^£0 sr^,
CERI 89-257
Revised April, 1990
PRO ^
WORKSHOP ON WATER QUALITY-BASED
PERMITTING, ENFORCEMENT, AND
TOXICITY REDUCTION EVALUATIONS
Boston, MA - May 22-23, 1990
Charlotte, NC - June 5-6, 1990
New Brunswick, NJ - June 19-20, 1990
Baltimore, MD - July 17-18, 1990
Madison, Wl - August 7-8, 1990
Denver, CO - September 18-19, 1990

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CXO1-!
WORKSHOP ON WATER QUALITY-BASED
PERMITTING, ENFORCEMENT, AND
TOXICITY REDUCTION EVALUATIONS
MAY 1990
U.S EPA Region 8 Library
80C-L
999 181 h St., Suilc 500
Denver, CO 80202-2466

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TABLE OF CONTENTS
Section 1
Recent Developments in the NPDES Program 	 1
Section 2
Concepts of Biological Testing 	 11
Section 3
Permit Limit Development	 75
Section 4
Compliance Monitoring and Enforcement	 107
Section 5
Basic Permitting Principles/Introduction to Toxicity Reduction Evaluations (TRE)
Introduction to TREs
TREs in Permitting and Enforcing Process
TRE Available Guidance	 125
Section 6
TRE Municipal and Industrial Protocols	 145
Section 7
Toxicity Identification Evaluation (TIE) Overview	 201
Section 8
Guidelines and Review Criteria for TRE Plans	 217
Section 9
TRE Case Studies and Summary 	 223

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Section 1
Recent Developments in the NPDES Program
-i-

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RECENT DEVELOPMENTS
WATER QUALITY ACT OF 1987
-	SLUDGE
-	STORMWATER
-	304(L)
COMBINED SEWER OVERFLOWS
REVISIONS TO THE TSD
DOMESTIC SEWAGE STUDY REGULATIONS
REVISIONS TO THE TSD
SCHEDULE:
« REGIONAL REVIEW (END 10/10/89)
•	WILLIAMSBURG GROUP REVIEW
-States	-Interest Groups
-Industry -Environmental Groups
•FEDERAL REGISTER ANNOUNCEMENT
•	BLUE RIBBON PANEL REVIEW
•	FINAL DOCUMENT
FINAL DOCUMENT AVAILABLE SUMMER '90
REVISIONS TO THE TSD
MAJOR CHANGES:
CHAPTER 1: New documentation
Correlations
CHAPTER 2: Acceptable ambient concentrations
Legal basis for AACs
CHAPTER 3: Streamlined procedures
Screening
Bioconcentration
-3-

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REVISIONS
TO THE TSD
MAJOR CHANGES:
CHAPTER 4:
Mixing Zones
Bioconcentration
CHAPTER 5:
More user-friendlyl
CHAPTER 6:
Permitting and Enforcement
Principles
ifii

Surface Water Toxics Control Program
Activities Conducted Under §304(1)
-4-

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Interrelationship of Waters Listed Under
Section 304(1) of the Clean Water Act
MINI LIST

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r
304(1)
"C LIST'
• 879 POINT SOURCES
° 625 INDUSTRIALS
-	134 METAL FINISHING
-	94 PULP & PAPER
-	55 NATURAL GAS
-	22 ORGANIC CHEMICAL
-	21 PETROLEUM REFINING
o 240 MUNICIPALS
° 14 FEDERAL FACILITIES
• ICSs REQUIRED
Individual Control Strategies
•	All known water quality problem waters impaired by
§307(a) toxics due entirely/substantially to point source
discharges require an ICS (Short List)
•	An ICS is a NPDES permit plus documentation (i.e.,
TMDLs/WLAs and other rationale)
•	An ICS is to produce a reduction in the discharge of
§307(a) toxic pollutants and achieve State WQ standards
within 3 years
•	Effluent toxicants, ammonia, and chlorine must also be
controlled by permits und6r other CWA authorities
304(l)-Changes to 40 CFR 122.44
SUBSECTION (d) - WATER QUALITY
STANDARDS AND STATE REQUIREMENTS
-	WATER QUALITY -BASED PERMIT LIMITS
FOR SPECIFIC TOXICANTS
-	WHOLE EFFLUENT TOXICITY LIMITS
WHERE NECESSARY TO 1
STATE WATER QUALITY !
V.
-6-

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Section 122.44(d)(l)(i)
ALL POLLUTANTS THAT CAUSE.
HAVE THE REASONABLE POTENTIAL
TO CAUSE OR CONTRIBUTE TO AN
EXCURSION ABOVE A WATER QUALITY
STANUARU MUST BE CONTROLLED
-	Includes narrative and numerical
criteria
-	Reflects EPA's approach to water
quality-based permitting
Section 122.44(d)(1)(H)
STATES MUST USE VALID PROCEDURES
TO DETERMINE WHETHER A DISCHARGE
CAUSES, HAS THE REASONABLE
POTENTIAL TO CAUSE. OR CONTRIBUTES
TO AN EXCURSION-"	
ACCOUNT FOR:
-	existing controls
-	variability
-	species sensitivity
-	dilution (where allowed)
Section 122.44(d)(l)(iii)
NPDES PERMITS MUST INCLUDE
EFFLUENT LIMITATIONS FOR EVERY
POLLUTANT THAT CAUSES, HAS THE
REASONABLE POTENTIAL TO CAUSE
OR CONTRIBUTES TO AN EXCURSION
ABOVE A_NUMERIC WATER QUALITY '
"CRITERION	'
-7-

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Section 122.44(d)(l)(iv)
NPDES PERMITS MUST INCLUDE WHOLE
EFFLUENT TOXICITY LIMITATIONS
WHEN A DISCHARGE CAUSES. HAS THE
REASONABLE POTENTIAL TO CAUSE.
OR CONTRIBUTES TO AN EXCURSION
ABOVgLA SXAT-F NUMERIC CRITFRTflN-
TOITWHOLE EFFLUENT TOXICITY
Section 122.44(d)(l)(v)
WHEN A DISCHARGE CAUSES. HAS THE
REASONABLE POTENTIAL TO CAUSE.
OR CONTRIBUTES TO AN EXCURSION
ARflVF A STATF. NAPRATTW. WATff.H
8UALITY CRITERION. THEPERMIT
UST CUMAlNXlMITATIONS ON~
WHOLE EFFLUENT TOXICITY *
- unless chemical specific limitations
are demonstrated to be sufficient
to achieve all applicable water
quality standards
Section 122.44(d)(l)(vi)
WHERE AN ACTUAL OR PROJECTED
EXCURSION ABOVE A WATER QUALITY
CRITERION IS ATTRIBUTABLE TO A
—PARTICULAR POLLUTANT FOR WHICH
«	THE STATE HAS NOT ADOPTED WATER
gUALITY CRITEgH)N. THE PERMIT MUST
«	CONTAIN WATER QUALITY-BASED
*	EFTLIJEN TLlMlTATlORS^O^CQIjTRQX,
. THE POLLUTANT OF CONCERN
-8-

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Section 122.44(d)(l)(vi)
THREE OPTIONS:
(1)	CALCULATE NUMERIC CRITERION
FOR THE POLLUTANT;
(2)	USE EPA's WATER QUALITY ^ H ^
CRITERION FOR THE POLLUTANT; OR
(3)	ESTABLISH EFFLUENT LIMITATIONS
ON AN INDICATOR PARAMETER
W
Section 122.44(d)(l)(vi)
IF AN INDICATOR PARAMETER IS USED,
FOUR PROVISIONS MUST BE MET:
/ (a) The permit must identify which
pollutants are intended to be controlled
by the indicator parameter;
(b) The fact sheet must set forth the
J basis for the limit;
is.. ^ (c) The permit must require monitoring
to show continued compliance with the
A water quality standards; and
(d) The permit must contain a reopener
clause allowing for changes (as needed) to
achieve water quality standards
Section 122.44(d)(l)(vii)
ALL WATER QUALITY-BASED EFFLUENT
LIMITATIONS MUST ADHERE TO
TWO PRINCIPLES:
(1) THE EFFLUENT LIMITATIONS MUST
BE DERIVED FROM AND COMPLY WITH
ALL APPLICABLE WATER QUALITY
STANDARDS
(2) THE EFFLUENT LIMITATIONS MUST
BE CONSISTENT WITH THE ASSUMPTIONS
AND REQUIREMENTS OF APPLICABLE
WASTELOAD ALLOCATIONS (IF
AVAILABLE)
-9-

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Section 2
Concepts of Biological Testing
-11-

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Section 122.44(d)(l)(v)
WHEN A DISCHARGE CAUSES. HAS THE
REASONABLE POTENTIAL TO CAUSE,
OR CONTRIBUTES TO AN EXCURSION
ABOVE A STATE NARRATIVE WATER
gUALITT CRITERION, THE PERMIT
MUST CONTAIN LIMITATIONS ON
WHOLE EFFLUENT TOXICITY
- unless chemical specific limitations
are demonstrated to be sufficient
to achieve all applicable water
quality standards
-13-

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LIMITATIONS OF
CHEMICAL-SPECIFIC APPROACH
•	All toxicants in complex wastewaters may
not be known (and therefore not limited).
*	Measurements of many individual toxicants
can be expensive (e.g., organic chemicals).
•	The bioavailability of the toxicants is not
assessed.
*	The interactions between toxicants (e.g.,
additivity, antagonism) are not measured.
LIMITATIONS OF
WHOLE-EFFLUENT APPROACH
•	Properties of specific chemicals are not
assessed (e.g., bioaccumulation).
•	Effluent toxicity treatability data are
lacking (engineers are more familiar with
designing systems to treat specific
chemicals).
•	WET testing at the point of discharge does
not account for changes in toxicity down-
stream due to chemical/physical conditions
(e.g., pH changes, photolysis, etc.).
-14-

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RESPONSE
ESTIMATORS
• ACUTE TOXICITY
—	SHORT TERM EXPOSURE
—	96 HOUR TEST
—	LETHALITY
• CHRONIC TOXICITY
-- LONG TERM EXPOSURE
— LIFE CYCLE TEST
~ LETHALITY
GROWTH
REPRODUCTION
TEST PROCEDURES
° TYPE OF TEST
-	STATIC, RENEWAL, FLOW-THRU
© NUMBER OF CONCENTRATIONS
-	100, 50, 25, 12.5, 6.25, 0
® NUMBER OF SPECIES
-	3 RECOMMENDED
• NUMBER OF ORGANISMS
-	20 RECOMMENCED
« DUPLICATIONS
® AGE OF ORGANISMS
-15-

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TEST PROCEDURES
(CONTINUED)
• TEST CONDITIONS
-	DURATION
ACUTE 24, 48, 96 h
CHRONIC 7, 14, 20 d
-	EFFECT MEASURED
MORTALITY, HATCHING, REPRODUCTION, GROWTH
-	CHAMBER LOADING
Sg 1L FLOW-THRU
0.8g 1L STATIC
-	D.O. > 40% SATURATION
NPDES COMPLIANCE MONITORING INSPECTOR TRAINING MODULE: BIOMONITORING
FIGURE 2-L TYPICAL EFFLUENT CONCENTRATIONS LSED IN TESTING
-16-

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LC --THE POLLUTANT CONCENTRATION AT WHICH 50 PERCENT
50 OF THE TEST ORGANISMS ARE KILLED WITHIN A SPECIFIED
TIME
NOEC--THE HIGHEST EFFLUENT CONCENTRATION AT WHICH NO
OBSERVED EFFECT WILL OCCUR AT CONTINUOUS EXPOSURE
TO TEST ORGANISMS
Response Curve for a Hypothetical Effluent
Acute Toxicity Test
PERCENT EFFLUENT
-17-

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CHRONIC DATA
The effects of an effluent on the growth and survival of inland silverside (Menidia
beryl1ina) larvae during an on site 7-day exposure. Test salinity was 32 , temperature 25
± 2°C. Three replicates of fifteen larvae were used in each concentration.
Effluent
Concen-
tration
(%>
Survival
(%)
Mean Dry
Weight/Larvae
(± S.D.)
(nig)
Mean
Salinity
(± S.D.)
(°/ )
* ' OO'
Mean
Temperature
(± S.D.)
CO
Mean
D.O.
(± S.D.)
(mg/1)
Control
(Brine +
D.I.)
87
0.963 ± 0.02
34
±
0
25.3
+
0.5
4.6
+
0.2
1.0
80
1.071 ± 0.08
34
+
0
25.4
+
0.3
4.4
+
0.1
3 . 2
58**
0.988 + 0.18
34
+
0.4
25.3
+
0.1
4.4
+
0.2
10.0
33*
0.737 ± 0.34
34
+
0
25.2
+
0.4
3.7
+
0.2
32 . 0
0*
	
35
+
0
24.3
+
0.4
6.8
+
0.3
50.0
0*
	
34
+
0
23.9
±
0.4
4.9
+
1. 1
* Significantly different from the control (a = 0.05).
** High variability between 3 replicates, not significantly different from the control.
Response Curve for a Hypothetical Effluent
Chronic Toxicity Test
z:
o
i—
o
3
O
o
a:
n.
hi
(Y1
o
i—
O
D
O
Li I
rt:
LlJ
u
fK
LJ
CL
I 10
6 2 12 5 25
PERCENT EFFLUENT
-18-

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ACUTE VALUE = IC50 = a point estimate
(a calculated value)
CHRONIC VALUE = NOEC =
= ICP =
a concentration endpoint
a point estimate
(a calculated value)
TOXIC UNITS (TU)
X TU = 100
LC50 OR NOEC
TUa = Toxic Units ACUTE
TUC = Toxic Unite CHRONIC
-19-

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EPA TOXICITY TESTS
SPECIES
Ceriodaphinia dubia
Daphina magna
Daphina pulex
Pirr.ephales promelas
Oncorhynchus myksis
14 Alternate Species
ACUTE
ENDPOINT
mortality
DURATION
48-96 hours
V
SPECIES
Pimeohales promelas
Pimeohales promelas
Ceriodaphinia dubla
Seienastrum
caprlcomutum
SHORT-TERM CHRONIC
ENDPOINT	DURATION
larval growth
and survival
larval survival and
teratogenicity
survival and
reproduction
growth
7 days
7 days
7 days
(approx)
96 hours
MARINE AMD KSTUARINE
Rapid
Chronic Toxicity Testa
Test Duration
Teat Salinity Rang*
Taat Bndpointa
Arbacia punctulata
Sea urchin
Champia parvula
Red macroalgae
Mvsidopsis bahia
Mysid
CvprJnodon varleoatus
Larval sheepshead minnow
Cyprilnp^on varleqatus
Embryo-larval
Sheepshead minnow
Menidia bervllina
Inland silverside
Total=1.5 hours
1	hour sperm exposure
20 min. fertilization
Total=7-9 days
2	day exposure
5-7 day incubation
Total=7 days
7 day exposure
Total=7 days
7 day exposure
Total«»9 day
9 day exposure
Total=7 days
7 day exposure
30 o/oo
28-32 o/oo
20-30 o/oo
20-32 o/oo
5-32 o/oo
5-32 O/OO
Fertilization
Cystocarp
production
(Fertilization)
Growth,
Survival,
Fecundity
Growth,
Survival
Percent Hatch,
Survival,
Percent
abnormality
Growth,
Survival
-20-

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United States	Environmental Monitoring and	EPA 600 4-85 013
Environmental Protection Support Laboratory	March 1985
Agency	Cincinnati OH 45268
Researcn and Development
Methods for
Measuring the Acute
Toxicity of Effluents to
Freshwater and
Marine Organisms
(Third Edition)

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TABLE 1
Summary of Recommended Test Conditions for Daphnid
(.Daphnia pulex and Daphnia magna) Acute Toxicity Test
1.	Temperature:
2.	Light quality:
3.	Light intensity:
4.	Photoperiod:
5.	Test chamber size:
6.	Test solution volume:
7.	Age of test organisms:
8.	Neonates/test chamber:
9.	Replicate
chambers/concentration:
10.	Total number of organisms
per concentration:
11. Feeding regime:
12. Aeration:
13. Dilution water:
14. Test duration:
15. Effects measured:
20 ± 2°C
Ambient laboratory illumination
10-20 jiiE/m 2/s (50-100 ft-c) (ambient lab
levels)
8-16 hours light/24 hours
100 mL beaker or equivalent
50ml/replicate (loading and D.O. must be met)
1-24 hr (neonates)
10
20
Feeding not required during first 48 hr. For
longer tests, feed every other day beginning on
the third day.
None, unless D.O. falls below 40% of
saturation, at which time start gentle, single-
bubble, aeration.
Receiving water or other surface water, ground
water, or svnthetic water: hard water for
' •>
Daphnia magna; moderately hard or soft water
for Daphnia pulex
Screening test - 24 h (static test)
Definitive test - 48 h (static test)
Mortality - no movement of body or
appendages on gentle prodding (LC50)
See Methods for Measuring the Acute Toxicity of Effluents m Freshwater and Marine Organisms. EPA/600/4-85/013, March 1985.
-22-

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TABLE 1
Summary of Recommended Test Conditions for Daphnid
(Daphnia pulex and Daphnia magna) Acute Toxicity Test
16. Test acceptability:	Control survival of 90% or greater
See Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms. EPA/600/4-85/013, March 1985.
-23-

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TABLE 2
Summary of Recommended Test Conditions for Mysid
(Mysidopsis bahia) Acute Toxicity Test
1.	Temperature:
2.	Light quality:
3.	Light intensity:
4.	Photoperiod:
5.	Test chamber size:
6.	Test solution volume:
7.	Age of test organisms:
8.	Organisms/test chamber:
9.	Replicate
chambers/concentration:
10.	Total number of organisms
per concentration:
11. Feeding regime:
12.	Aeration:
13.	Dilution water:
14.	Test duration:
15.	Effects measured:
20 + 2°C
Ambient laboratory illumination
10-20 AiE/m 2/s (50-100 ft-c) (ambient lab
levels)
8-16 hours light/24 hours
250 mL beaker or equivalent
200ml/replicate (loading and D.O. restrictions
must be met)
1-5 days
10
20
Two drops of concentrated brine shrimp nauplii
suspension twice daily (approx. 100
nauplii/mysid)
None, unless D.O. falls below 40% of
saturation, at which time start gentle, single-
bubble, aeration.
Natural seawater, or synthetic salt water
adjusted to 20 ppt salinity
Screening test - 24 h (static test)
Definitive test - 48 h (static test);
48-96 h (flow-thru)
Mortality - no movement of body or
appendages on gentle prodding (LC50)
Se; Methods for Measuring the Acute Toxicity of Effluents to Freshwatcrand Marine Organisms. EPA/600/4-85/013, March 1985.
-24-

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TABLE 2
Summary of Recommended Test Conditions for Mysid
(Mysidopsis bahia) Acute Toxicity Test
16. Test acceptability:	Control survival of 90% or greater.
See Methods for Measuring the Acute Toxicity of Effluents to Freshwaterand Marine Organisms. EPA/600/4-85/013, March 1985.
-25-

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TABLE 3
Summary of Recommended Test Conditions for Fathead Minnow
{Pimephales promelas) Acute Toxicity Test
1.	Temperature:
2.	Light quality:
3. Li"ht intensity:
C	*
4.	Photoperiod:
5.	Test chamber size:
6.	Test solution volume:
7.	Age of test organisms:
S. Number of fish/test chamber:
9.	Replicate
chambers/concentration:
10.	Total number of organisms
per concentration:
11. Feeding regime:
12.	Aeration:
13.	Dilution water:
14.	Test duration:
15.	Effects measured:
16.	Test acceptability:
20+ 2°C
Ambient laboratory illumination
10-20 |LtE/m 2/s (50-100 ft-c) (ambient lab
levels)
8-16 hours light/24 hours
1 L beaker or equivalent
0.75 L/replicate (loading and D.O. restrictions
must be met)
1-90 days
10
20
Feeding not required first 96 h
None, unless D.O. falls below 40% of
saturation, at which time start gentle, single-
bubble, aeration.
Receiving water, other surface water, ground
water, or soft synthetic water
Screening test - 24 h (static test)
Definitive test - 48 h (static test);
48-96 h (flow-thru)
Mortality - no movement (LC50)
Control survival of 90% or greater.
See Methods for Measuring the Acute Toxicity of Effluents to F-eshwnterand Marine Organisms, EPA/600/4-85/013. March 1985.
-26-

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TABLE 4
Summary of Recommended Test Conditions for Silverside
{Menidia spp.) Acute Toxicity Test
1.	Temperature:
2.	Light quality:
3.	Light intensity:
4.	Photoperiod:
5.	Test chamber size:
6.	Test solution volume:
7.	Age of test organisms:
8.	Organisms/test chamber:
9.	Replicate
chambers/concentration:.
10.	Total number of organisms
per concentration:
11.	Feeding regime:
12.	Aeration:
13.	Dilution water:
14.	Test duration:
20 + 2°C (northern latitudes)
25 + 2°C (southern latitudes)
Ambient laboratory illumination
10-20 /iE/m2/s (50-100 ft-c) (ambient lab
levels)
8-16 hours light/24 hours
1 L beaker or equivalent
0.75 L/replicate (loading and D.O. restrictions
must be met)
1-90 days
10
20
Feeding not required first 96 h
None, unless D.O. falls below 40% of
saturation, at which time start gentle, single-
bubble, aeration.
Natural seawater, or synthetic salt water
adjusted to 25-30 ppt salinity
Screening test - 24 h (static test)
Definitive test - 48 h (static test);
48-96 h (flow-thru)
15. Effects measured:
Mortality - no movement (LC50)
See Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms. EPA/600/4-85/013, March 1985.
-27-

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TABLE 4
Summary of Recommended Test Conditions for Silverside
(Menidia spp.) Acute Toxicity Test
16. Test acceptability:	Control survival of 90% or greater.
See Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms. EPA/600/4-85/013, March 1985.
-28-

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&EPA Short-Term Methods for
Estimating the Chronic
Toxicity of Effluents and
Receiving Waters to
Freshwater Organisms
Second Edition

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Anatomy of female Daphnia pulex (De Geer), X70; A, antenna; BC,
brood chamber; H, heart; INT, intestine; L, legs; OV, ovary; P,
postabdomen; PC, postabdominal claw. (From Pennak, 1978).

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CERIODAPHNIA SURVIVAL AND REPRODUCTION
O SCOPE OF TEST-MEASURES THE CHRONIC TOXICITY OF WHOLE
EFFLUENTS TO THE CLADOCERAN, CERIODAPHNIA DUBIA. DURING
A 7-DAY, STATIC RENEWAL EXPOSURE.
O SUMMARY OF METHOD—CERIODAPHNIA ARE EXPOSED IN A STATIC
RENEWAL SYSTEM FOR 7 DAYS TO DIFFERENT CONCENTRATIONS
OF EFFLUENT OR RECEIVING WATER. TEST RESULTS ARE BASED ON
SURVIVAL AND REPRODUCTION. IF THE TEST IS CONDUCTED
£ PROPERLY, THE CONTROL ORGANISMS SHOULD PRODUCE THREE
BROODS OF YOUNG (10-35 NEONATES/BROOD) DURING THE 7-DAY
PERIOD.
O END POINT—CERIODAPHNIA MORTALITY AND REPRODUCTION
O TEST OBSERVATIONS-FIRST BROOD ON THIRD DAY IS 2-5 YOUNG,
THEREAFTER A BROOD OF 10-35 NEONATES IS RELEASED EVERY
36-48 HOURS.
-MORTALITY AND/OR NUMBER OF YOUNG PER ADULT FEMALE ARE
RECORDED DAILY.

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CERIQDAPHN1A SURVIVAL AND REPRODUCTION
(Continued)
O NUMBER AND AGE OF ORGANISMS-60 NEONATES < 4-H OLD
AT THE BEGINNING OF THE TEST (10 ORGANISMS/CONCENTRATION
+ CONTROL).
O TERMINATION OF TEST-ALL OBSERVATIONS MUST BE COMPLETED
IN 7 DAYS (± 2-H) AFTER INITIATION OF TEST.
I
** O DATA ANALYSIS-TOTAL NUMBER OF YOUNG PER ADULT FEMALE IS
DETERMINED UNTIL DEATH OR END OF TEST, WHICHEVER COMES
FIRST. CALCULATE MEAN NUMBER OF YOUNG PER ADULT FEMALE
FOR EACH CONCENTRATION TO PROVIDE A COMBINED MEASURE
OF THE TOXICANT'S EFFECT ON MORTALITY AND REPRODUCTION.
O CALCULATIONS-SEE CHRONIC METHODS MANUAL FOR DETAILED
DISCUSSION.

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TABLE 7
Summary of Recommended Test Conditions for
Ceriodaphnia Survival and Reproduction Test
1.	Test type:
2.	Temperature:
3.	Light quality:
4.	Light intensity:
5.	Photoperiod:
6.	Test chamber size:
7.	Test solution volume:
8.	Renewal of test concentrations:
9.	Age of test organisms:
10.	Neonates/test chamber:
11.	Replicate
chambers/concentration:
12.	Neonates/concentration:
13.	Feeding regime:
14.	Aeration:
15.	Dilution water:
Static renewal
25°C ± l'C
Ambient laboratory illumination
10-20 /xE/m2/s (50-100 ft-c) (ambient lab
levels)
16 hours light, 8 hours darkness
30 mL beakers or equivalent
15 mL/replicate (loading and D.O. restrictions
must be met)
Daily
Less than 24 hour; all released within a 8-h
period
1
10
10
Feed 0.1 mL each of YCT and algal suspension
per test chamber daily
None
Moderately hard synthetic water is prepared
using Millipore Milli-QR or equivalent
deionized water and reagent grade chemicals or
20% DMW.
16. Effluent concentrations:
Minimum of 5 and a control
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.
EPA/600/4-89/001, March 1989.
-33-

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TABLE 7
Summary of Recommended Test Conditions for
Ceriodaphnia Survival and Reproduction Test
17.	Dilution factor:
18.	Test duration:
19.	Endpoint:
20.	Test acceptability:
21. Sampling requirement:
Approximately 0.3 or 0.5
Until 60% of control females have three broods
(may require more or less than 7 days).
Survival and reproduction
80% or greater survival and an average of 15
or more young/surviving female in controls. At
least 60% of surviving females in controls
should have produced their third brood.
For on-site tests, samples are collected daily,
and used within 24 h of the time they are
removed from the sampling device. For off-
site tests, a minimum of three samples are
collected, and used on days 1-2, 3-4, and 5-7.
22. Sample volume required:
1 L/day
See Short-Term Methods for Estimatine ihe Chronic Toxicity of Effluents and Receiving Waters to FreshwaterOrganisms.
EPA/600/4-89/001, March 1989.
-34-

-------
TABLE 2. DATA FROM CERIODAPHNIA EFFLUENT TOXICITY TEST
Total No. Most Young
Effl Day 	Replicate	 Live Live By Any
Cone
No.
A
B
c
D
E
F
G
H
I
J
Young
Adults
One Adult

3
0
0
0
0
0
X
0
0
0
0
0
9
0

4
2
2
4
0
6
•
2
6
1
4
27
9
6
Cont
5
9
2
9
0
9
-
0
6
2
9
46
9
9

6
5
6
9
6
0
-
3
5
11
10
55
9
11

7
6
8
5
10
1
_
3
8
10
3
54
9
10


77
TT
T7
TT
TT
T
~T
TT
TT
TT
ITT


3
0
0
0
0
0
0
0
0
0
0
0
10
0

4
2
4
l
0
4
2
2
3
2
2
22
10
4
1.0X
5
9
5
2
0
8
13
2
8
2
2
51
10
13

6
6
3
6
2
8
8
10
0
6
6
55
10
10

7
5
3
12
4
3
8
9
0
13
12
69
10
13


77
TT
TT
1
TT
31
23
TT
TT
77
197


3
0
0
0
0
0
0
0
0
0
0
0
10
0

4
2
1
2
4
4
8
2
0
4
3
30
10
8
3.OX
5
3
4
2
9
11
6
2
4
8
6
55
10
11

6
10
8
6
9
0
3
6
10
8
3
63
10
10

7
10
12
10
2
0
6
11
8
0
6
65
10
12


TT
75"
tt
TT
TT
TS
TT
77
TT
TT
2TT

3
0
0
0
0
0
0
0
0
0
3
3
10
3

4
2
4
2
2
0
l
2
0
4
2
19
10
4
6.OX
5
2
8
0
2
4
2
2
4
6
8
38
10
8

6
9
2
2
3
10
6
8
6
2
4
52
10
10

7
10
2
3
6
12
13
9
11
9
4
79
10
13


TT
TT
1
TT
TT
77
TT
TT
TT
TT
lTT

3
0
0
0
0
0
0
0
0
0
0
0
10
0

4
0
4
2
2
2
0
1
2
1
4
18
10
4
12.OX
5
1
8
0
2
3
3
0
2
3
0
22
10
8

6
8
4
8
3
10
10
5
10
6
7
71
10
10

7
11
5
7
6
10
12
10
11
8
7
87
10
12


TT
tt
T7
TT
TT
TT
TT
TT
TT
TT
1ST

3
0
0
0
0
0
0
0
0
0
0
0
10
0

4
X
0
0
0
X
X
0
X
0
0
0
6
0
25.OX
5
-
X
1*
X
-
-
X
•
X
X
1
1
1

6
-
-
-
-
-
m
-
-
-
m
0
0
0

7
-
-
-
•
m
•

•
m

0
o
0



IT
1
~T
T
T
IS
"7
T
IS
T


x » Dead adult, no young produced before death.
lx * Dead adult; one young produced before death.
Note: Days 1 and 2 are not Included because young were not produced until
the third day. Adult mortality was not recorded for days 1 and 2.
-35-

-------
Fathead minnow: adult female
(right).
(left) and breeding male

-------
FATHEAD MINNOW (PIMEPHALES PROMELAS)
LARVAE
O SCOPE OF TEST-ESTIMATE THE CHRONIC TOXICITY OF WHOLE
EFFLUENTS TO FATHEAD MINNOW LARVAE IN A 7-DAY, STATIC
RENEWAL TEST.
O SUMMARY OF METHOD-LARVAE (PREFERABLY < 24-H OLD) ARE
ij EXPOSED IN A STATIC RENEWAL SYSTEM FOR 7 DAYS TO
DIFFERENT CONCENTRATIONS OF EFFLUENT TEST RESULTS ARE
BASED ON THE SURVIVAL AND GROWTH (INCREASE IN WEIGHT) OF
THE LARVAE.
O END POINT-MORTALITY AND GROWTH (CHANGE IN WEIGHT) OF
THE LARVAE.
O NUMBER AND AGE OF ORGANISMS-150-300 PER TEST; THREE
SEPARATE SPAWNS; <24-H OLD.

-------
FATHEAD MINNOW (PIMEPHALES PROMELAS)
LARVAE(Continued)
O TERMINATION OF TEST-AFTER 7 DAYS THE LARVAE IN EACH
CHAMBER ARE PRESERVED AS A GROUP. LARVAE ARE RINSED,
OVEN DRIED (100° C FOR >2-H) AND WEIGHED (0.1MG).
UJ
00
1
O DATA ANALYSIS-MORTALITY AND MEAN WEIGHT OF LARVAE FOR
EACH CONCENTRATION ARE COMPARED TO CONTROLS AND
STATISTICALLY SIGNIFICANT (P = 0.05) DIFFERENCES DETERMINED.
O CALCULATIONS-SEE CHRONIC METHOD MANUAL FOR DETAILED
DISCUSSION.

-------
TABLE 5
Summary of Recommended Test Conditions for Fathead Minnow
(Pimephales promelas) Larval Survival and Growth Test
1.
Test type:
Static renewal
2.
Temperature:
25°C ± 1°C
3.
Light quality:
Ambient laboratorv illumination
4.
Light intensity:
10-20 \lE/m2/s (50-100 ft-c) (ambient lab
5.	Photoperiod:
6.	Test chamber size:
7.	Test solution volume:
8.	Renewal of test concentrations:
9.	Age of test organisms:
10.	Larvae/test chamber:
11.	Replicate
chambers/concentration:
12.	Larvae/concentration:
13.	Feeding regime:
14. Cleaning:
levels)
16 hours light, 8 hours darkness
500 mL beakers or equivalent
250 mL/replicate (loading and D.O. restrictions
must be met)
Daily
Newly hatched larvae (less than 24 hours old)
15 larvae/chamber (minimum 10)
4 (minimum of 3)
60 larvae/concentration (minimum 30)
Feed 0.1 mL newly hatched (less than 24-h old)
brine shrimp nauplii three times daily at 4-h
intervals or, as a minimum, 0.15 mL twice
daily, 6 h between feedings (at the beginning of
the work day prior to renewal, and at the end
of the work day following renewal). Sufficient
larvae are added to provide an excess. Larvae
are not fed during the final 12 h of. the test.
Siphon daily, immediately before test solution
renewal
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.
EPA/600/4-89/001, March 1989.
-39-

-------
TABLE 5
Summary of Recommended Test Conditions for Fathead Minnow
(Pimephales promelas) Larval Survival and Growth Test
15. Aeration:
16. Dilution water:
None, unless D.O. falls below 40% of
saturation. Rate should not exceed 100
bubbles/min.
Moderately hard synthetic water is prepared
using Millipore Milli-QR or equivalent
deionized water and reagent grade chemicals or
20% DMW
17.	Effluent concentrations:
18.	Dilution factor:
19.	Test duration:
20.	Endpoints:
21.	Test acceptability:
22.	Sampling requirement:
Minimum of 5 and a control
Approximately 0.3 or 0.5
7 days
Survival and growth (weight)
80% or greater survival in controls; average dry
weight of surviving controls equals or exceeds
0.25 mg
For on-site tests, samples are collected daily,
and used within 24 h of the time they are
removed from the sampling device. For off-
site tests, a minimum of three samples are
collected, and used on days 1-2, 3-4, and 5-7.
23. Sample volume required:
2.5 IVday
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.
EPA/600/4-89/001, March 1989.
-40-

-------
TABLE 6
Summary of Recommended Test Conditions for Fathead Minnow
(Pimephales promelas) Embryo-Larval Survival and Teratogenicity Test
1.	Test type:
2.	Temperature:
3.	Light quality:
4.	Light intensity:
5.	Photoperiod:
6.	Test chamber size:
7.	Test solution volume:
8.	Renewal of test concentrations:
9.	Age of test organisms:
10.	Larvae/test chamber:
11.	Replicate
chambers/concentration:
12.	Larvae/concentration:
13.	Feeding regime:
14.	Aeration:
Static renewal
25"C ± 1°C
Ambient laboratory illumination
10-20 AtE/m2/s (50-100 ft-c) (ambient lab
levels)
16 hours light, 8 hours darkness
150-500 mL beakers or equivalent
70-200 mL/replicate (loading and D.O.
restrictions must be met)
Daily
Less than 36 hour old embryos
15 larvae/chamber (minimum 10)
4 (minimum of 3)
60 larvae/concentration (minimum 30)
Feeding not required
None, unless D.O. falls below 40% of
saturation. Rate should not exceed 100
bubbles/min.
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and ReceivingWaters to Freshwater Organisms.
EPA/600/4-89/001, March 1989.
-41-

-------
TABLE 6
Summary of Recommended Test Conditions for Fathead Minnow
(.Pimephales promeias) Embrvo-Larval Survival and Teratogenicity Test
15. Dilution water:
16.	Effluent concentrations:
17.	Dilution factor:
18.	Test duration:
19.	Endpoint:
20.	Test acceptability:
21.	Sampling requirement:
Moderately hard synthetic water is prepared
using Millipore Milli-QR or equivalent
deionized water and reagent grade chemicals or
20DMW. The hardness of the test solutions
must equal or exceed 25 mg/L (CaC03) to
ensure hatching.
Minimum of 5 and a control
Approximately 0.3 or 0.5
7 days
Combined mortality (dead and deformed
organisms)
80% or greater survival in controls
For on-site tests, samples are collected daily,
and used within 24 h of the time they are
removed from the sampling device. For off-
site tests, a minimum of three samples are
collected, and used on days 1-2, 3-4, and 5-7.
22. Sample volume required:
2.5 L/day
See Short-Tern Methods for Estimating the Chronic Tcacitv of Effluents and ReceivineWaters to Freshwater Organisms.
EP.V600/4-89/001. March 1989.
-42-

-------
TABLE 2. SUMMARY OF SURVIVAL AND GROWTH DATA OF FATHEAD MINNOW LARVAE EXPOSED FOR
SEVEN DAYS TO SOOIUM PENTACHLOROPHENATE
Proportion of
Test NaPCP Survival In	Mean Ave Dry Wgt (mg) In	Hean
Cone. Cone. Replicate Chambers	Prop.	CVa Replicate Chambers	Dry Wgt CV
No. (ug/L) A B C D	Surv.	(X) A B C	D («g) (%)
1
Control
1.0
1.0
0.9
0.9
0.95
6
0.711
0.662
0.718
0.767
0.714
6
2
32
0.6
0.8
1.0
0.8
0.85
12
0.646
0.626
0.723
0.700
0.674
7
3
64
0.9
1.0
1.0
1.0
0.975
5
0.669
0.669
0.694
0.676
0.677
2
4
128
0.9
0.9
0.8
1.0
0.90
9
0.629
0.680
0.513
0.672
0.624
12
5
256
0.7
0.9
1.0
0.5
0.775
29
0.650
0.558
0.606
0.508
0.580i>
11
6
512
0.4
0.3
0.4
0.2
0.325*
29
0.358
0.543
0.488
0.495
0.471«>
17
'Coefficient of variation (stanaard deviation X 100/nean).
bS1gn1f1cantly different from control (P • 0.05).

-------
ALGAL (SELENASTRUM CAPRICORNUTUM)
GROWTH TEST
SCOPE OF TEST-MEASURES THE CUR.»NIC TOXICITY OF WHOLE
EFFLUENT TO THE FRESH WATER ALGA, SELENASTRUM
CAPRICORNUTUM, DURING A 4-DAY STATIC EXPOSURE.
SUMMARY OF METHOD-A SELENASTRUM POPULATION IS EXPOSED
IN A STATIC SYSTEM TO A SERIES OF EFFLUENT CONCENTRATIONS
FOR 96-H. THE RESPONSE OF THE POPULATION IS MEASURED IN
TERMS OF CHANGES IN CELL DENSITY (CELL COUNTS PER ML),
BIOMASS, CHLOROPHYLL CONTENT, OR ABSORBANCE. BY
EXTENDING THE TEST TO 14 DAYS, IT MAY BE USED TO MEASURE
THE ALGAL GROWTH POTENTIAL OF WASTEWATERS AND
SURFACE WATERS.
-SELENASTRUM IS A UNICELLULAR COCCOID GREEN
ALGA.

-------
ALGAL (SELENASTRUM CAPRICORNUTUM)
GROWTH TEST (Continued)
END POINT-CELL COUNTS OR CHLOROPHYLL CONTENT OR
TURBIDITY (LIGHT ADSORBANCE). REGARDLESS OF THE METHOD,
TEST SOLUTIONS SHOULD BE CHE ck6D UNDER THE MICROSCOPE
TO DETECT ABNORMALITIES IN THE CELL SIZE OR SHAPE.
METHODS OF MEASURING THE END POINT
(1)	COUNTING-AUTOMATIC PARTICLE COUNTER; COUNTS EACH
CELL PASSING THROUGH AN APERATURE BY MEASURING A
CHANGE IN VOLTAGE. CELLS MAY BE MANUALLY COUNTED,
MINIMUM OF 400 CELLS PER REPLICATE.
(2)	CHLOROPHYLL CONTENT-CHLOROPHYLL MAY BE MEASURED
IN-VIVO FLUOROMETRICALLY OR SPECTROPHOTOMITRICALLY.
FLUOROMETRIC MEASUREMENTS ARE RECOMMENDED
BECAUSE OF SIMPLICITY AND SENSITIVITY.

-------
ALGAL (SELENASTRUM CAPRICORNUTUM)
GROWTH TEST(Continued)
o METHODS OF MEASURING THE END POINT
(3)	TURBIDITY(ABSORBANCE)— A SPECTROPHOTOMETER IS USED
TO DETERMINE THE TURBIDITY, OR ABSORQ^NCE. OF THE
CULTURES AT A WAVE LENGTH 750 NM. VOLUME, SIZE, AND
PIGMENTATION OF THE ALGAE CAN AFFECT THE RESULTS;
CALIBRATE TO ESTABLISH A RELATIONSHIP BETWEEN
ABSORBANCE AND CELL DENSITY.
(4)	BIOMASS—ALGAL GROWTH POTENTIAL RESULTS MAY BE
EXPRESSED AS A MG DRY WEIGHT ORGANIC MATTER/LITER.
BIOMASS CAN BE CALCULATED FROM CELL COUNTS AND
MEAN CELL VOLUMES, OR CAN BE MEASURED DIRECTLY BY
GRAVIMETRIC METHODS.
O NUMBER AND AGE OF CELLS-10,000 CELLS/ML/DILUTION AND
REPLICATE FROM 4 TO 7 DAY STOCK.
o CALCULATIONS-SEE CHRONIC METHODS MANUAL FOR DETAILED
DISCUSSION

-------
TABLE 8
Summary of Recommended Test Conditions for the AJgal
(Selenastrum capricomutum) Growth Test
1.
Test type:
Static
2.
Temperature:
25 • C ± 10 C
3.
Light quality:
"Cool white" fluorescent lighting
4.
Light intensity:
86 ± 8.6 ME/m 2/s (400 ± 40 ft-c)
5.
Photoperiod:
Continuous illumination
6.
Test chamber size:
125 or 250 mL beakers or equivalent
7.
Test solution volume:
50 or 100 mL/replicate
8.
Renewal of test concentrations:
None
9.
Age of test organisms:
4 to 7 days
10.
Initial cell density:
10,000 cells/mL
11.
Replicate chambers/conc.:
3
12.
Shaking rate:
100 cpm continuous, or twice daily by hand
13.
Dilution water:
Algal stock culture medium without EDTA or
enriched surface water
14.
Effluent concentrations:
Minimum of 5 and a control
15.
Dilution factor:
Approximately 0.3 or 0.5
16.
Test duration:
96 h
17.
Endpoint:
Growth (cell counts, chlorophyll fluorescence,
absorbance, biomass)
18.
Test acceptability:
2 X 105 cells/mL in the controls; variability of
controls should not exceed 20%
19.
Sample volume required:
1 L (one sample for test initiation)
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms.
EPAJ600/4-89/001, March 1989.
-47-

-------
TABLE 4. SAMPLE OATA FROM ALGAL TOXICITY TEST WITH CAOHIUH CHLORIDE

Growth
Cells/mL
Percent
Toxlcmt
Response
O
O
o
Inhibition
Concentration
Cells/nt
(Logio)
of Growth
(uq Cd/L)
1000
lit)
0 (Control)
1209
3.082


1180
3.072
0

1340
3.127

5
1212
3.084


1186
3.074
3.4

1204
3.081

10
826
2.917


628
2.798
39.1

816
2.912

20
493
2.693


416
2.619
64.5

413
2.616

40
127
2.104


147
2.167
88.7

147
2.167

80
49.3
1.693


40.0
1.602
96.4

44.0
1.643

-48-

-------
United Slates
Environmental Protection
Agency
Environmental Monitoring and EPA 600 4-87 028
Support Laboratory	May 1988
Cincinnati OH 45268
Research and Development
SEPA Short-Term Methods for
Estimating the Chronic
Toxicity of Effluents and
Receiving Waters to
Marine and Estuarine
Organisms

-------
TABLE 13
Summary of Recommended Test Conditions
for Arbacia punctulata fertilization test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Light quality:
5.	Light intensity:
6.	Test vessel size:
7.	Test solution volume:
8.	Number of sea urchins:
9.	Number of egg and sperm cells
per chamber:
10.	Replicate
chambers/treatment:
11.	Dilution water:
12.	Dilution factor:
13.	Number of treatments/study:
14.	Test duration:
15.	Effects measured:
Static
30ppt ±2ppt
20°±1°C
Ambient laboratory light during test
preparation
10-20 /xE/m 2/s (50-100 ft.c.) (ambient lab
levels)
Disposable (glass) liquid scintillation vials
(20ml capacity), not pre-cleaned
5ml
Pooled sperm from four males and pooled eggs
from four females per test
About 2000 eggs and 5,000,000 sperm cells per
vial
4 (minimum of 3)
Uncontaminated source of natural seawater or
deionized water mixed with hypersaline brine
or artificial sea salts
0.3 or 0.5
Minimum of 5 effluent concentrations and a
control
1 hour and 20 minutes
Fertilization of sea urchin eggs
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-50-

-------
TABLE 13
Summary of Recommended Test Conditions
for Arbacia punctulata fertilization test
16. Test acceptability:	Control fertilization sperm:egg ratio of 70-90%.
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-51-

-------
5 mm
Figure 1. Life history of Chang) 1a parvula. Upper left: Size and decree of
branching 1n female branch tips used for toxicity tests. Fro« Thursby and
StMle, 1987.
-52-

-------
TABLE 14
Summary of Recommended Test Conditions
for Champia parvula sexual reproduction test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Photoperiod:
5.	Light quality:
6.	Light intensity:
7.	Test vessel size:
8.	Test solution volume:
9.	Dilution water:
10.	Dilution factor:
11.	Number of dilutions:
12.	Number of replicate Chambers
per treatment:
13.	Number of organisms
per test chamber:
14.	Test duration:
15. Effects measured:
Static
30ppt ±2ppt
22-24 "C
16 h light, 8 h dark
Cool white fluorescent lights
100 nE/m 2/s (500 ft.c.)
200 mL polystyrene cups, or 250 mL
Erlenmeyer flasks
100ml
30 parts per thousand salinity natural seawater,
or a combination of 50% (30 part per thousand
salinity) natural seawater and 50% (30 part per
thousand) salinity artificial seawater
0.3 or 0.5
At least 5 and a control
4 (minimum of 3)
5 female branch tips and 1 male plant
2-day exposure to effluent, followed by 5 to 7
day recovery period in control medium for
cystocarp development
Reduction in cystocarp production compared to
controls
See Short-Term Methods for Estimating the ChronicToxicitv of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-53-

-------
TABLE 14
Summary of Recommended Test Conditions
for Champia parvula sexual reproduction test
16. Test acceptability:	Control survival of 80% or greater, control
average cystocarp production of 10 or greater
per plant. (NOTE: plants fragmenting in lower
concentrations may indicate undue stress)
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-54-

-------
MATURE FEMALE, EGGS IN BROOD SAC
eyesto Ik
ontennule
antenna
carapace
pleopods
brood sac with
developing embryos
statocyst
telson
uropod'
oviducts with developing ova
Figure 3. Mature female M. bahia with eggs 1n oviducts and developing
embryos 1n the~brood sac. Above: lateral view. Below: dorsal
view. From Lussier, Kuhn, and Sewall, 1987.
-55-

-------
TABLE 12
Summary of Recommended Test Conditions for the
Mysidopsis Bahia Seven Day Survival, Growth, and Fecundity Test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Light intensity:
5.	Photoperiod:
6.	Test chamber:
7.	Test solution volume:
8.	Renewal of test solutions:
9.	Age of test organisms:
10.	Organisms/test chamber:
11.	Replicate
chambers/treatment:
12.	Source of food:
13.	Feeding regime:
14.	Cleaning:
15.	Aeration:
16.	Dilution water:
Static renewal
20ppt to 30ppt ± 2ppt
26 °-270 C
10-20 ME/m2/s (50-100 ft.c.) (ambient lab
levels)
16 h light, 8 h darkness, with phase in/out
period
8oz plastic disposable cups or 400ml glass
beakers
150ml/replicate cup
Daily
7 days
5
8
Artemia nauplii
Feed 150 24-h old Artemia nauplii per mysid
daily, half after test solution renewal and half
after 8-12 hours
Pipette excess food from cups daily
None, unless D.O. falls below 60% of
saturation, then gently in all cups
Uncontaminated source of natural seawater or
hypersaline brine
See Short-Term Methods for Estimating the ChronicToxicitv of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-56-

-------
TABLE 12
Summary of Recommended Test Conditions for the
Mysidopsis Bahia Seven Day Survival, Growth, and Fecundity Test
17.	Number of treatments/study:
18.	Dilution factor:
19.	Test duration:
20.	Effects measured:
21.	Test acceptability:
Minimum of 5 and a control
Approximately 0.3 or 0.5
7 days
Survival, growth, and egg development
Control survival of 80% or greater, control dry
weight of 0.2 mg/mysid or greater.
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and RcccivingWatcrs to Marine and Estuarinc Organisms.
EPA/600/4-87/028, May 1988.
-57-

-------
TABLE 9
Summary of Recommended Test Conditions for Sheepshead
Minnow Cyprinodon variegatus Larval Survival and Growth Test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Light quality:
5.	Light intensity:
6.	Photoperiod:
7.	Test chamber size:
8.	Test solution volume:
9.	Renewal of test concentrations:
10.	Age of test organisms:
11.	Larvae/test chamber:
12.	Replicate
chambers/concentration:
13.	Source of food:
14.	Feeding regime:
15. Cleaning:
Static renewal
20ppt to 32ppt ± 2ppt
25°C ± 2°
Ambient laboratory illumination
10-20 jiiE/m 2/s (50-100 ft-c) (ambient lab
levels)
14	hours light, 10 hours darkness
300ml - 1L beakers or equivalent
250-750ml/replicate (loading and D.O.
restrictions must be met)
Daily
Newly hatched larvae (less than 24 hours old)
15	larvae/chamber (minimum 10)
4 (minimum of 3)
Newly hatched Anemia nauplii (less than 24
hours old)
Feed once a day 0.10g wet weight Artemia
nauplii per replicate on Days 0-2; feed 0.15g
wet weight Artemia nauplii per replicdte on
Days 3-6
Siphon daily, immediately before test solution
renewal
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988
-58-

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TABLE 9
Summary of Recommended Test Conditions for Sheepshead
Minnow Cyprinodon variegatiis Larval Survival and Growth Test
16.	Aeration:
17.	Dilution water:
18.	Effluent concentrations:
19.	Dilution factor:
20.	Test duration:
21.	Effects measured:
22.	Test acceptability:
None, unless D.O. falls below 60% of
saturation, then aerate all chambers. Rate
should be less than 100 bubbles/minute
Uncontaminated source of natural seawater,
hypersaline brine, or artificial seawater mixed
with deionized water
5 and a control
Approximately 0.3 or 0.5
7 days
Survival and growth (weight)
Control survival of 80% or greater, and average
control average dry weight of 0.6 mg or greater
or, if preserved, 0.5 mg or greater.
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Rcccivine Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988

-------
TABLE 10
Summary of Recommended Test Conditions for Sheepshead Minnow
Cyprinodon variegatus Embryo Larval Survival and Teratogenicity Test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Light quality:
5.	Light intensity:
6.	Photoperiod:
7.	Test chamber size:
8.	Test solution volume:
9.	Renewal of test concentrations:
10.	Age of test organisms:
11.	Embryos/test chamber:
12.	Replicate
chambers/concentration:
13.	Embryos per concentration:
14.	Feeding regime:
15.	Aeration:
16.	Dilution water:
Static renewal
5ppt to 32ppt ± 2ppt
25°C ± 2°
Ambient laboratory illumination
10-20 /xE/m 2/s (50-100 ft-c) (ambient lab
levels)
14	hours light, 10 hours darkness
500ml
400ml (minimum of 250 ml)
Daily
less than 24 hours old
15	embryos/chamber (minimum 10)
4 (minimum of 3)
60 (minimum of 30)
Feeding not required
None, unless D.O. falls below 60% of
saturation
Uncontaminated source of natural seawater,
hypersaline brine, or artificial seawater mixed
with deionized water
17. Effluent concentrations:
5 and a control
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-60-

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TABLE 10
Summary of Recommended Test Conditions for Sheepshead Minnow
Cyprinodon variegatiis Embryo Larval Survival and Teratogenicity Test
18.	Dilution factor:
19.	Test duration:
20.	Effects measured:
Approximately 0.3 or 0.5
9 days
Percent hatch; percent larvae dead or with
debilitating morphological and/or behavior
abnormalities such as: gross deformities,
curving spine, disoriented, abnormal swimming
behavior; surviving normal larvae from original
embryos
21. Test acceptability:
Control survival of 80% or greater.
See Short-Term Methods for Estimating the ChronicToxicitv of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-61-

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TABLE 11
Summary of Recommended Test Conditions for the Inland
Silverside Menidia beryllina Larval Survival and Growth Test
1.	Test type:
2.	Salinity:
3.	Temperature:
4.	Light quality:
5.	Light intensity:
6.	Photoperiod:
7.	Test chamber size:
8.	Test solution volume:
9.	Renewal of test concentrations:
10.	Age of test organisms:
11.	Larvae/test chamber:
12.	Replicate
chambers/concentration:
13.	Source of food:
14.	Feeding regime:
15.	Cleaning:
16.	Aeration:
Static renewal
5ppt to 32ppt ± 2ppt
25°C ± 2°
Ambient laboratory illumination
10-20 jLiE/m 2/s (50-100 ft-c) (ambient lab
levels)
14	hours light, 10 hours darkness
300ml - 1L containers
250-750ml/repIicate (loading and D.O.
restrictions must be net)
Daily
7-11 days post hatch
15	larvae/chamber (minimum 10)
4 (minimum of 3)
Newly hatched Artemia nauplii
Feed 0.10g wet weight Artemia nauplii per
replicate on days 0-2; Feed 0.15g wet weight
Artemia nauplii per replicate on days 3-6
Siphon daily, immediately before test solution
renewal and feeding
None, unless D.O. falls below 60% of
saturation, then aerate all chambers. Rate
should be less than 100 bubbles/minute
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-62-

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TABLE 11
Summary of Recommended Test Conditions for the Inland
Silverside Menidia beryllina Larval Survival and Growth Test
17.
Dilution water:
Uncontaminated source of natural seawater or
hypersaline brine mixed with deionized water
18.
Effluent concentrations:
at least 5 and a control
19.
Dilution factor:
Approximately 0.3 or 0.5
20.
Test duration:
7 days
21.
Effects measured:
Survival and growth (weight)
22.
Test acceptability:
Control survival of 80% or greater, control
average dry weight (for 7 day old larvae) of 0.5
mg or greater, or, if preserved, 0.43 mg or
greater.
See Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receivine Waters to Marine and Estuarine Organisms.
EPA/600/4-87/028, May 1988.
-63-

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CHRONIC DATA
The effects of an effluent on the growth and survival of inland silverside (Menidia
bervllina) larvae during an on site 7-day exposure. Test salinity was 32 °/00, temperature
25 ± 2 °C. Three replicates of fifteen larvae were used in each concentration.
Effluent

Mean Dry
Mean
Mean
Mean
Concen-

Weight/Larvae
Salinity
Temperature
D.O.
tration
Survival
(± S.D.)
(± S.D.)
(± S.D.)
(± S.D.)
(%)
(%)
(mg)
(°/ )
V / 00'
(•C)
(mg/1)
Control
(Brine +
D.I.)
87
0.963
+
0.02
34
+
0
25.3
+
0.5
4.6
+
0.2
1.0
80
1.071
+
0.08
34
+
0
25.4
+
0.3
4.4
+
0.1
3.2
58**
0.988
+
0. 18
34
+
0.4
25.3
+
0.1
4.4
+
0.2
o
o
33*
'0.737
+
0. 34
34
+
0
25.2
+
0.4
3.7
+
0.2
32 . 0
0*

— -
—
35
+
0
24.3
+
0.4
6.8
+
0.3
50.0
0*

—
—
34
+
0
23.9
+
0.4
4 . 9
+
1.1
* Significantly different from the control (a = 0.05).
** High variability between 3 replicates, not significantly different from the control.
VARIABILITY IN
TOXICITY TEST RESULTS
SPECIES USED
STRAIN OR SOURCE OF TEST ORGANISMS
CONDITION OR HEALTH OF THE ORGANISMS
TEST CONDITIONS
•	TEMPERATURE
•	D.O.
9 FOOD
•	WATER QUALITY
•	ETC.
-64-

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ACUTE RELATIVE SENSITIVITY
(ranked from most sensitive = 1, to least = 3)
SPECIES	AMMONIA CHLORINE DDI COPPER
Fathead	3	3 3	3
Daphnia	2	111
Rainbow
Trout	1	2 2	2
PRECISION - the degree of reproducibility of test results
2 TYPES OF PRECISION:
Intra-laboratory precision (within) - the ability of trained
lab personnel to repeatedly obtain consistent results
when performing the same test on the same species
using the same toxicant.
Inter-laboratory precision (round-robin) - how reproducible
a method is when conducted by a large number of
laboratories performing the same test on the same
species using the same toxicant.
-65-

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CV = Coefficient of Variation
A measure of precision
standard deviation X 100
mean
EXAMPLES OF PRECISION DATA
Cerioriaphnia
Sheepshead minnow
Intra-lah
(CV%)
41.13 NAPCP
27.6 Copper
33.3 SDS
Inter-lab
(CV%)
41.1	NAPCP
29.0 NaCI
44.2	Effluent
-66-

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Tabic 1-6
Intra-iaboratory Precision of Chronic Whole Effluent Taocity Tea Methods
Test NOEC
Method Ranse
Mean
1C25
CVf%1
Mean
[C50
CVfW
Comoound
Water
Used
Cvcrinodoo varientes
Survival A Growth






50 ; 0 ug/l ,
0.5 - 1.0
31 - 125 uy\ ,
1.3 - 2.5 mg/1
70
1.5
300.4
2.2
41.8
31.4
33.0
27 6
130
1.9
396.9
16
408
31.8
19.2
35.3
Copper
SDS
Copper
SDS
AS
AS
NS
NS
Embryo larvaJ survival A teratogenicity
200 - 240 \i%!\
2.0 - 1.0 ugrt
EQO
202
1.9
2.3
35
EC50
233.5
11.7
2.5
2.9
Copper
SDS
AS
AS
Merndia bervilina - Survival A Growth






31-125 ug/l"
1.3 - 0 ug/l
209.9
1.3
43.7
43.2
340.8
1.9
50.7
9.4
Copper
SDS
NS
NS
Mvndooos bahia -Survival. Growth A Fecundity






<0.3 - 5 0 ug$
63 - 125 ug/l
5.7
138.3
35.0
18.0
6.9
185.8
47.8
5.8
SDS
Copper
NS
NS
Arbacia ouncnilata -Fcrtilizanoo






5.3 - 12.5 ug/l#*
1.2 - 3.3 mg/1 „
<6.1 - 24.4 ug/l
0.9 - 1.8 mg/1
23J
1.7
22.9
Z54
54.6
29.7
41.9
30.3
45.7
2.4
33.3
3.2
47.9
23.3
46.4
33.1
Copper
SDS
Copper
SDS
AS
AS
NS
NS
Chamoia oarvula - RcDroducaoo






0.5 - 1.0 ug/l* ..
0.09 - 0.48 mg/1
0.93
230
63
60.9
1.4
0.3
38.6
24.1
Copper
SDS
AS/NS
AS/NS
Pimenhaies ommelan - Survival A Growth
128- 256ug/l* ,
0.0U - 0.013 mg/1
Embryo larval survival A teratogenicity
0.011 - 0.013 mg/1
CeraxbotmiadiitNa - Reoroduciim


L£1
0.0068
1.51
62
4 U
NAPCP2
Cadmium
Cadium
Diquat
FW
FW
FW
FW
0.10 - 0 JO mg/1 *
0.22
41.13
0.3
27.9
NAPCP
FW
Seierurtm eanncnmimim- <* hour Survival






0.49- 10.0 ug/l***
0.23
102.6
0-38
75.6
Cadmium
chloride
FW
Notes
" Difference of 1 tea concentration AS-artificial seawater
"" Difference of 2 tea concentrations NS-naturai seawater
•** Difference of 4 test concentrations FW-freshwater
7 Sodium dodecyi sulfate
2 Sodium pentachlorophenol
-67-

-------
Table 1 - 7
Summary of tntcr-lab Variability Data [39. 42~44)
Chroruc.
1.
Test Method
Cvorinodon variecatui
NO EC Ranse
CV<1) CIC251
CV (10501

7 day growth 4 survival
1 - 3.2% effluent*
44.2%
56.9%
2.
PvmcDhaies oromelas




7 day growth & survival
<3 0 - 6.0 mg/1
potassium chromate
31.0
290
3.
Ceriodaohnia dubia



4
7 day reproduction
Ceriodaohnia dubia
0.25 - O.JQ mg/1
NAPCP®
41.1
27.9

7 day reproduction
6 - 12% effluent
—
—
5.
Ceriodaohnu dubia




7 day reproduction
<0.25 - 1.0 g/1
NaCl
29.0
40.0
Acute:

Toxicant

CV (LC50^
6.
Cvonnodoc vanczatus




96 hour static
96 hour flow-through
96 hour static
96 hour flow-through
endosulfan
endosulfan
silver nitrate
silver nitrate

37.7%
46.2
34.6
50.1
7
Mvsdoons bahia




% hour static
96 hour flow-through
96 hour static
96 hour flow-through
endosulfan
endosulfan
silver nitrate
silver nitrate

59.5%
51.9
26.6
22J
Notes
y CV-coefficientof variation
. NAPCP-Sodium penuchlorophenol
This represents a difference of one exposure concentration
— Data unavailable
-68-

-------
Table 1-4
Precision of Inorganic Analysis
at the Lov End of the Measurement Detection Range
Ana.lyte	No. of Labs	CV (%)
Aluminum
37
43
Cadmium
63
66
Chromium
72
40
Copper
86
36
Iron
78
38
Lead
64
46
Manganese
55
129
Mercury
76
79
Silver
50
18
Zinc
62
118
This data is taken from reference EPA/600/4-79/029, Revised
March 1983. Data provided to ERL-Duluth by W.Peltier.
Table 1-5
Precision Ranges for Organic Chemical Analysis
Chemical
Benzene
4 Chlorobenzenes
Ethyl benzene
Toluene
2 3 Halocarbons
4 Halocarbons
11 Phenols
Benzidine
3,3-Dichlorozidine
6 Pthalate esthers
3 Nitrosamines
24 Organochlorine
Pesticides and PCBs
No. CV
Labs ill
20 31-64
% Data EPA Document
Discarded Referenced
20
20
20
17
16
17
22
16-29
40-50
20-45
38-64
38-69
>12-45
10
¦>
->
20
->
22
1 9
¦>
600/S4-84-064
600/S4-84-064
600/S4-84-044
600/S4-84-062
600/S4-84-056
600/S4—84-051
600/S4-84-061
16 PNAs
16-91
600/S4-84-063
-69-

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Table 1-5 continued
Nitrobenzene
Isophorone	? 26-60	?	600/S4-B4-018
2,6-Dinitrotoluene
2,4-Dinitrotoluene
5 Haloethers	20 32-53	?	600/S4-84-039
8 Chlorinated	20 26-41	25	600/S4-84-039
hydrocarbons
Table 1-6
Precision of Non-metal Inorganic Analyses
Over the Measurement Range
Lab
Parameter
CV (%) Ranae
17
Alkalinity
4.9-14
>20
Residual chlorine
13-25
16
Ammonia nitrogen
15-58
6
Kjeldahl nitrogen, total
38-41
15
N03 nitrogen
17-61
6
Total P
25-40
53
BOO
15-33
58
COD
6.9-34
21
TOC
4.6-70
Data taken from EPA-600/4-79-020 (revised 1983).
-70-

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COMPLEX EFFLUENT TOXICITY
TESTING PROGRAM
8 Streams - 84 Monitoring Locations
RESULTS:
•	Ambient toxicity directly correlated to
stream population impact*
•	Effluent toxicity from single source
directly correlated to stream impact
* Stream population also affected
by temperature, D.O., pH, etc.
COMPARISON OF AMBIENT TOXICITY AND IMPACT
83 STATIONS ON 0 STREAMS IN THE U.S.
IMPACT PREDICTED
IMPACT PRESENT
68
NO MPACT PREDICTED
MP ACT PRESENT
NO IMPACT PREDICTED
NO MP ACT PRESENT 4'8%
MPACT PREDICTED 3 6%
NO MPACT PRESENT
-71-

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NORTH CAROLINA STUDY
43 Point Source Discharge Sites
No Inatream Toxicity Predict
Impact Noted 50*
Inatream Toxicity Predicted
Impact Noted _
65%
No Inatream Toxicity Predicted
No Impact Noted 23%
Inatream Toxicity Predicted
No Impact Noted -jy
SALT WATER STUDY
79 Ambient Stations and 4 Dischargers
PREDICTED AMBIENT TOXICITY
6% NO TOXICITY OBSERVED
NO AMBIENT TOXICITY PREDICTED
TOXICITY OBSERVED
5%
PREDICTED AMBIENT TOXICITY
TOXICITY OBSERVED ^ ^
NO AMBIENT TOXICITY PREDICTED
NO TOXICITY OBSERVED
-72-

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TEST CONDUCTED AT CENTRAL LABORATORY
NO. OF LABORATORIES CONDUCTING SHORT-TERM CHRONIC TESTS
COST RANGE ($)
DAPHNIDS
MINNOW
MINNOW
AGPT

LARVAL
EMBRYO

201-500
4
2
1
5
501-800
2
1
2
2
801-1100
2
3
4
2
1101-1400
7
2
1
2
1401-1700
2
4
—
—
1701-2000
—
1
4
--
2001-2300
1
—
1
«
3101-2600
—
1
1
-
TOTAL LABS
18
1
13
11
MEAN COST
1067
1220
1605
737
COST RANGE
300-2200
400-2500
400-2500
200-1400
TOXICITY TESTING COSTS
TEST
48-HOUR DAPHNIA ACUTE (STATIC)
96-HOUR FATHEAD MINNOWS ACUTE (STATIC)
24-HOUR ETT SCREENING TEST
9 6-HOUR ETT DEFINITIVE (FISH)
96-HOUR ONSITE FLOW-THROUGH
7-DAY CERIODAPHNIA CHRONIC
7-DAY FATHEAD MINNOW CHRONIC
21-DAY DAPHNIA CHRONIC
COST
$400-800
$500-900
$100-200
$500-900
$5,000-7,000
$1,100-1,800
$1,200-1,800
$S,000-7,000
-73-

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The following are the cost estimates associated with the current capabilities
of the toxicity testing center. These cost are per treatment and as such can
be applied equally to effluents and receiving waters (a typical effluent test
has 5 treatments).
These cost are based on a "one-time" testing routine. When multiple tests
are arranged for large projects the costs will be less per treatment. After the
initial year of operation, we will have a better handle on the true cost of
running the testing center. At that time the cost estimates will be revised,
and should come down.
Species
Cost per
Treatment
Champia, algal
Laminaria, algal
Sea urchin
Mysid-96 hr
Mysid-7 day
Ampelisca, sediment
Bivalve-48 hr
Fish-96 hr
Fish- 7 day
300.00
300.00
300.00
425.00
725.00
725.00
425.00
425.00
725.00
-74-

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Section 3
Permit Limit Development
-75-

-------
Objective
Ensure that effluents do
not cause or contribute
to exceedances of the
water quality standards
in the receiving water
Toxicity is inversely
proportional to
wet test endpoints
(NOEC, LC )
50
TOXIC UNITS (TU)
100
x TU = 	
LC OR NOEC
OU
TU. - Toxic Units ACUTE
d
TU. ¦ Toxic Units CHRONIC
o
-77-

-------
Effluent Cone.
-78-

-------
Effluent Cone.
Time
General Approach
1.	Gather information
2.	Determine applicable WQS
3.	Evaluate which parameters need limits
4.	Determine wasteload allocations (WLAs)
5.	Determine long-term average effluent
concentrations (LTAs)
6.	Determine permit limits
-79-

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Step 1 - Gather information
Effluent data.
Receiving water data
Step 2 - Determine applicable WQS
Narrative standard
No acute toxicity within or outside
mixing zone
Interpretation: 0.3 TUa
No chronic toxicity outside mixing zone
In terpre ta tion: 1.0 TU
* Also consider chemicals which do not
have numeric standards
-80-

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DURATION
MAGNITUDE \ FREQUENCY
CMC = 0.3 TUA  1Hr/3 yrs
CCC = 1.0 TUC © 4 DAYS/3 yrs
WLA IS A QUANTIFICATION OF AN AMBIENT
TARGET. IT IS SEPARATE FROM A PERMIT LIMIT
DOSE RESPONSE CURVE
ACUTE
w

<*>
7,
100
o
90
PL.


70
£
50
8
&
30
w

cu
10

0
PERCENT EFFLUENT
-SI-

-------
IX
120
no
'00
125
90
ao
u 70
I
i 60
as
57
SO
40
X
40
20
10
12
0- 11- 21- 31- 41- .51- 61- 71- 81- SI-
10 20 .30 .40 50 .00 . 70 .80 90 1.0
LC1/LCS0 Ratio
Step 2 (cont.)
Numeric standards
For most parameters, use values
in WQS
For ammonia and most metals, use
equations in WQS
(need receiving water data)
-82-

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Step 2 (cont.)
Also note duration and frequency for each WQS
• Most pollutants and toxicity
Acute 1 hr. avg. not to be exceeded
more than once every 3 years
Chronic 4 day avg. not to be exceeded
more than once every 3 years
• Pesticides, PCBs, Silver:
Review guidance before using
statistical approach
Step 3 -
Evaluate which parameters need limits
(a) Determine dilution factors
(b) Choose evaluation method
and evaluate need for limit
-83-

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Step 3 (cont.)
When is a limit needed?
If effluent "will cause,
have the reasonable potential
to cause, or contribute to"
an exceedance of the WQS
after mixing (if allowed).
Step 3 (cont.)
(a) Determine acute and chronic dilution
factors
Acute WQS applied:
•	near outfall, if
rapid initial mixing
•	otherwise at end-of-pipe
(i.e. dilution factor =1)
Chronic IVOS applied:
a at edge of mixing zone
-84-

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(A) When plant's water source is the
receiving stream
Qc
d.f.
Q
(B)	When plant's water source is not the
receiving stream
Qs + Q
d.f. .	-
e
(C)	Qe = effluent design flow
Qs = receiving stream design flow
1Q Acute
10
7Q10 Chronic
Step 3 (cont.)
To determine minimum dilution factors
near outfall or at edge of mixing zone,
use worst-case conditions.
•	Simple dilution calculations
(rivers and streams)
•	Field studies
•	Computer modeling
-85-

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Step 3 (cont.)
(b) Choose evaluation method and
evaluate need for limits
"Standard"
Actual
Concentration
Effluent
Receiving
Water
WLA
(multiply
by dilution
factor)
WQS
compare
compare
Effluent Cone.
(divide
by dilution
factor)
Receiving water
conc. after mixing
(IWC or RWC)
Traditional
Timesaving
Step 3 (cont.)
Toxicity - Convert units on effluent
before doing comparison
Acute No. of TUa's = 100
in effluent LC50(in %)
for most sensitive species
Note.- Calculation only works if
LCcn i 100% effluent
50
-86-

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Step 3 (cont.)
Acute (cont.)
•	LC50 = 100%
?
Effluent concentration < WQS * DFa
1 TU < 0.3 TU * DF
a	a a
Need DFa > 3.3 before no limit
is needed.
•	As LC50 decreases, need even greater
dilution.
Step 3 (cont.)
Acute (cont.)
Evaluating need for limit if LC50 > 100%
(1)	Significant mortality, but < 50%
® End-of-pipe (DFa= 1) Need limit always
• Near outfall	Depends on DFg and
degree of mortality
(2)	Mortality not significantly different from
control (= "no acute toxicity")
No limit needed
-87-

-------
Step 3 (cont.)
• Chronic
•	If have chronic data
No. of TU 's in effluent = 100/NOEC (in %)
C
•	If have only acute data
No. of TUc's in effluent = [100/LCso(in %)] * ACR
where ACR = acute/chronic ratio
= 10 if no chronic data available
Step 3 (cont.)
Traditional Method:
Acute
End-of-pipe
Near outfall
Chronic
Edge of mixing zone
If answer in either or both
approaching no, need limit.
-88-
Effluent Cone.
DF
WQS
Effluent Cone.
Effluent Cone.
DF_
<	WQS
(acute)
?
<	WQS
(acute)
Effluent Cone. '
DF
< WQS
(chronic)
cases is no, or is

-------
Step 3 (cont.)
Timesaving Method:
Acute
End-of-pipe
(DF = 1)
Near outfall
Effluent Cone. < WQS * DF
IWLA}
Effluent Cone. < WQS
(acute)
Effluent Cone. < WLA,
Chronic
Edge of mixing zone
Effluent Cone. < WLA
If answer is no, or is approaching no, need limit.
Approach
Step 4
Step 5
WLA
a
WLA
c
I I
L.T.A.
a
L.T.A.
c

Step 6
Limits
WLA Requirements
Required treatment
Select limiting
L.T.A.
Calculate limits
•	daily maximum
•	monthly average
-89-

-------
Step 4 - Determine acute and chronic
waste/oad allocations
If used traditional approach in Step 3:
(1)	Multiply acute WQS by acute dilution
factor to get WLAa.
(2)	Repeat using chronic WQS and chronic
dilution factor to get WLAc .
If used timesaving approach in Step 3,
use acute and chronic WLAs from that step.
Step 4 - (cont.)
Units Conversion for Toxicity WLAs:
•	Have acute WLA in units of TU , and
chronic WLA in units of TU . a
c
•	Need units same for later comparison
averages.
•	Convert units: Acute/Chronic Ratio (ACR)
WLA (in TUa) * ACR = WLAac(in TUC)
where ACR = acute chronic ratio
LCJin .%)
		 for effluent
NOEC (in %)
(assume ACR » 10 if no data)
-90-

-------
Step 5 - Determine long-term average effluent
concentrations (LTAs) required to meet WLAs.
Information needed:
•	Acute and chronic WLAs (from Step 4)
•	Probability that effluent will meet WLA
(99%)
•	Effluent variability
Step 5 (cont.)
Effluent Variability
Coefficient of variation (CV)
= measure of how far values
vary about mean
= standard deviation / mean
-91-

-------
CV = 10
CV = 0 6
A A . . /

^ vv
N/
Relationship Between Wasteload Allocation and Long-Term
Average for Different Coefficients of Variation
Step 5 (cont.)
Coefficient of variation (CV)
•	If variability mostly related to production
and expect similar variability in future,
can use existing effluent data. *
•	Otherwise use CV = 0.6
•	Find s and x for , x2 , etc.
CV = s/x
-92-

-------
What are eqns. in Steps 5 and 6 accomplishing?
• Transformation from "real world" into
"log world" where statistics are valid,
then back into "real world."
• Transition from 1-hr and 4-day average
WQS to daily max. and monthly ave.
permit limits.
BOD5 FREQUENCY DISTRIBUTION
PLANT C
ISO
100
80
o
CONCENTRATION IN mg/l
I I
u «•
18
-93-

-------
METAL PRIORITY POLLUTANT
FREQUENCY DISTRIBUTION
PLANT A
>•
o
z
Ui
3
o
ui
DC
IL
140
120
100
80
•0
40
20
0
n
2=n=
0.4
t.2 2.0
2.8
CONCENTRATION IN mg/l
Step 5 (cont.)
Find acute LTA required to meet WLAa
(WLAac for toxicity)
g Know: WLA a
CV
Z
B Calculate: a =
2
a
P
LTAa =
-94-
from Step 4
from effluent data, or 0.6
2.326 for 99% probability
of meeting WLA
\/In (CV2+ 1)
In (CV2+ 1)
In (WLAa ) - Za
e (jj, + 0.5 a2)

-------
Step S (cont)
Find chronic LTA required to meet WLAj.
WLAC from Step 4
CV
same as for acute
LTA calculation
Calculate: /x 4 = In (WLAC ) - Z \f In
1 + (e °2-l)
4 J
jj. = fj.4 - 0.5 a ^+ 0.5 In
1 + (ea ¦ 1)
4 —1
LTA- = e (A + °-5 a1)
Step 5 (cont.)
Select the lowest of
LTAa and LTA, for use
a	C
in Step 6.
-95-

-------
TIME
(Days)
z
o
I-
<
DC
H
Z
HI
O
z
o
o
>-
z
UJ
1L
IL
HI
WLA
— A
	MAN DM.V
WLA.
AVO montiKv
TM« ITA(*llh«f LT^or 11(^1, gtvsn (Ms CV, w« tcNtvt t«lh WlAo
10 11 12 13 14 IB
(Days)
-96-

-------
Permit Limit Derivation
Technology-Based Water Quality-Based
Best plant performance	V\^S
WLA = "effluent standard"
1
LTA effluent concentration	LTA effluent conc. needed to
I1	meet WLA
I
Set permit limits (monthly ave. and Same as for technology-based
daily max.) to ensure LTA is	limits
achieved.	Hi
I	I
Required technology-based	Required water quality-based
performance for all plants	performance for one plant
III |J
I
jent
I
Step 6 -Determine monthly average and daily
maximum permit limits required to meet LTA.
Information needed:
•	LTA (from Step 5)
•	Effluent variability (CV)
•	Probability that effluent will meet LTA
daily max. 99%
mo. ave. 95%
•	No. of samples to be taken per month
-97-

-------
Step 6 (cont.)
Derive daily maximum permit limit.
I Know: LTA lowest one from Step 5
CV same as Step 5
Z 2.326 (for 99%)
a 1
L same as Step 5
„2 J
B Calculate: fi = In (LTA) - 0.5 
-------
Parameter: COPPER
Acute Wasteload Allocation (WLA, acute) —
Chronic Wasteload Allocation (WLA, acute) -
Coefficient of variation (CV) of effluent =
Monthly sampling frequency required in permit
68.000 ug/L
42.000 ug/L
0.600
1.000 samples per month
Back calculate the long term average (LTA)
that will meet both of the above WLAs:
est s
est u.
est u.
LTA
4d
Id
Lowest LTA
Acute
0.555
NA
2.930
21.834
Chronic
0.555
3.055
2.944
22.152 ug/1
21.834 ug/1
Using the lowest LTA and CV from above, derive the
Maximum Daily and Monthly Average permit limits
n=l
n=4
Percentile Basis
95th %ile 99th %ile
Percentile Basis
95th %ile 99th %ile
est s2
0.307
0.307
est u
2.930
2.930
Maximum Daily =
46.613
68.000
Monthly n =
1.000
1.000
est s2.n
0.307
0.307
est u.n
2.930
2.930
Monthly Average =
46.613
68.000
46.613 68.000 ug/L
33.895 41.396 ug/L
-99-

-------
Timesavers for Steps 5 and 6:
Tables - Permit Writer's Guide
(p. 12)
Computer Spreadsheet
-100-

-------
Table 5-1
Back Calculation of Long Term Average Wasteload

WLA multipliers

cv
[ 0.5 c
e
2-zc]


95th
percentile
99th
percentile
acute
0.1
0.853
0.797

CM CO
d o
0.736
0.644
0.643
0.527
1 TA 1AM A [ 0.5 O2-Z0]
LTAa = WLAa»e
0.4
0.571
0.440

0.5
0.6
0.514
0.468
0.373
0.321
where a2 = In [ CV2 + 1 ],
0.7
0.432
0.281
z = 1.645 for 95th percentile occurrence probability, and
0.8
0.9
0.403
0.379
0.249
0.224
z = 2.326 for 99th percentile occurence probability
1.0
0.360
0.204

1.1
0.344
0.187

1.2
0.330
0.174

1.3
0.319
0.162

1.4
0.310
0.153

1.5
0.302
0.144

1.6
0.296
0.137

1.7
0.290
0.131

1.S
0.285
0.126

1.9
0.231
0.121

2.0
0.277
0.117



WLA multipliers

CV
[ 0.5 o
e
i2 - Z o4 ]


95th
99th
chronic

percentile
percentile



(4-day average)
p O
to ^
0.922
0.853
0.891
0.797

0.3
0.791
0.715

0.4
0.736
0.643
1 TA Ullt [ 0.5 c42 - z 
-------
Table 5-2
Calculation of Permit Limits
CV
LTA multipliers
[zc-
e
0.5 o2 ]
95th
percentile
99th
percentile
0.1
1.17
1.25
0.2
1.36
1.55
0.3
1.55
1.90
0.4
1.75
2.27
0.5
1.95
2.68
0.6
2.13
3.11
0.7
2.31
3.56
0.8
2.48
4.01
0.9
2.64
4.46
1.0
2.78
4.90
1.1
2.91
5.34
1.2
3.03
5.76
1.3
3.13
6.17
1.4
3.23
6.56
1.5
3.31
6.93
1.6
3.38
7.29
1.7
3.45
7.63
1.8
3.51
7.95
1.9
3.56
8.26
2.0
3.60
8.55
Maximum Daily Limit
MDL ss LTA • e
' z a- 0.5 a2 ]
where a2 = ln[ CV2 + 1 ],
z =» 1.645 for 95th percentile occurrence probability, and
z = 2.326 for 99th percentile occurence probability
Average Monthly Limit
AML = LTA • e
[zon-0.5 on2]
where crn2 - In [ CV2 / n +1 ],
z = 1.645 for 95th percentile,
z = 2.326 for 99th percentile, and
n = number of samples/month
CV
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.3
1.9
2.0
LTA multipliers
;zan -0.530
1.17
1.36
1.55
1.75
1.95
2.13
2.31
2.48
2.64
2.78
2.91
3.03
3.13
3.23
3.31
3.38
3.45
3.51
3.56
3.60
1.12
1.25
1.38
1.52
1.66
1.80
1.94
2.07
2.20
2.33
2.45
2.56
2.67
2.77
2.86
2.95
3.03
3.10
3.17
3.23
-102-
1.08
1.17
1.26
1.36
1.45
1.55
1.65
1.75
1.85
1.95
2.04
2.13
2.23
2.31
2.40
2.48
2.56
2.64
2.71
2.78
1.06
1.12
1.18
1.25
1.31
1.38
1.45
1.52
1.59
1.66
1.73
1.80
1.87
1.94
2.00
2.07
2.14
2.20
2.27
2.33
1.03
1.06
1.09
1.12
1.16
1.19
1.22
1.26
1.29
1.33
1.36
1.39
1.43
1.47
1.50
1.54
1.57
1.61
1.64
1.68
99th
percentile
n=1 n=2 n=4 n=10 n=30
1.25
1.55
1.90
2.27
2.68
3.11
3.56
4.01
4.46
4.90
5.34
5.76
6.17
6.56
6.93
7.29
7.63
7.95
8.26
8.55
1.18
1.37
1.59
1.83
2.09
2.37
2.66
2.96
3.28
3.59
3.91
4.23
4.55
4.86
5.17
5.47
5.77
6.06
6.34
6.61
1.12
1.25
1.40
1.55
1.72
1.90
2.08
2.27
2.48
2.68
2.90
3.11
3.34
3.56
3.78
4.01
4 23
4 46
4.68
4.90
1.08
1.16
1.24
1.33
1.42
1.52
1.62
1.73
1.84
1.96
2.07
2.19
2.32
2.45
2.58
2.71
2.84
2.98
3.12
3.26
1.04
1.09
1.13
1.18
1.23
1.28
1.33
1.39
1.44
1.50
1.56
1.62
1.68
1.74
1.80
1.87
1.93
2.00
2.07
2.14

-------
Paraaeter: COPPER
Acute Wasteload Allocation (WLA.acute) =	18.00 uq/L
Chronic Wasteload Allocation (WLA.chronic) =	12.00 uo/L
Coefficient of variation (CV) of effluent =	0.60
Monthly samelino freouencv reauired in oennit =	4.00 saiaoles/iaonth
Back calculate the long term averape (LTA)
that Hill meet both of the above WLAs:
Acute Chronic
est.s
0.555
0.555
est u. 4d
NA
1.802
est u. Id
1.601
1.691
LTA
5.779
6.329 uq/L
Lowest LTA =	5.779 ua/L
Usino the lowest LTA and CV from above, derive the
Maximum Daily and Monthly Averaos oenait limits
Percentile Basis
95th X'ile 99th 7/ile
est	s2	0.307	0.307
est	u	1.601	1.601
Itaffluffi Dailv	=	12.339	18.000 uq/L
Month lv	n =	4.000
est	s2.n	0.086	0.086
est	u.n	1.711	1.711
Monthlv Averaoe =	8.972	10.958 ua/L
-103-

-------
Parameter: Toxicity
Acute Wasteload Allocation (WLA,acute) =
Chronic Wasteload Allocation (WLA,chronic) =
Coefficient of variation (CV) of effluent =
Monthly sampling frequency required in permit =
Back calculate the long term average (LTA) that win meet the above wla
Acute Chronic
3.66 TUc
1.32 TUc
0.60
4.00 samples/month
est S	0.555	0.555
est U, 4d	NA	-0.405
est U, Id	0.008	-0.516
LTA	1.175	0.696 TUc
Lowest LTA =	0.6 96 TUc
Using the lowest LTA and CV from above, derive the
Maximum Daily and Monthly Average permit limits
Percentile Basis
95th %'ile 99th Vile
est S2	0.307 0.307
est u	-0.516	-0.516
Maximum Daily =	1.486 2.168 TUc
Monthly n =	4.000
est s2,n	0.086 0.086
est U,n	-0.405	-0.405
Monthly Average =	1.081 1.320 TUc
-104-

-------
Fact Sheet
•	Results of evaluation showing
which parameters need limits
•	Determination of limits for each parameter
•	Cite method followed and source
•	Give values used:
•	Ambient data (for WQS)
® Max. effluent values and ACR
•	DF and DF
a	c
•	WLA and WLA
a	c
•	cv
•	No. of samples /month
•	Probabilities for calculating
LTA, daily max., and mo. avg.
Checklist of Data Needs
(for Parameters in WQS, Including Toxicity)
Essential Helpful
Concentrations
Max. values
Long-term data
Acute-chronic ratio
Receiving water
Hardness, pH, temp., salin.
Ambient pollutant conc.'s
Dilution factors





\r
I ^

|

I

-105-

-------
Section 4
Compliance Monitoring and Enforcement
-107-

-------
ENFORCEMENT PRINCIPLES
The same principles apply to whole-effluent toxicity enforcement
as enforcement of chemical-specific limits.
PERMITTEES are responsible for:
Attaining
Monitoring, and
Maintaining COMPLIANCE with their permits
(and for data quality)
REGULATORS are responsible for:
TRACKING COMPLIANCE with and
ENFORCING against violations of permit requirements
These principles and the integration of whole-effluent toxicity
control into the compliance monitoring and enforcement process
are embodied in the COMPLIANCE MONITORING AND
ENFORCEMENT STRATEGY FOR TOXICS CONTROL.
COMPLIANCE MONITORING
PROCESS
1.	Determining permit requirements;
2.	Tracking compliance and determining
violations;
3.	Reporting and placing the violations in
priority order; and
4.	Determining appropriate response.
-109-

-------
1. DETERMINING PERMIT
REQUIREMENTS
Communication between permit writer and enforcement is
vital
Permits must be enforceable (wording is important); permits
should be easy to track (difficult permits are more likely to
fall through the cracks!)
Determining permit requirements (or even enforcement order
requirements) may sound easy, but it isn't always. Permits may
be soundly written from a water-quality perspective, but difficult
to track (e.g., based on stream-flow, effluent flow, or even the
Spring thaw!). Others may just be poorly written:
CONSENT DECREE: WORDING IS
IMPORTANT
A Poorly Worded Example:
Beginning on the day that follows by two weeks the last day of
the next month following the date of entry of this decree, and
continuing thereafter on the corresponding day of each
subsequent month until the termination of the decree,	
shall prepare and transmit a monthly report for the second
previous month...
Another example of permit wording with significant import
from an enforcement standpoint is a limit which is not framed in
terms of daily (monthly) average and daily maximum (or weekly
average for POTWs) as required in 40 CFR 122.45.
When determining the statutory maximum penalty that may
be assessed for violation of a permit, it is important to know the
period of time represented by a given violation.
In the absence of a defined limit (in terms of days represented
by the reported value), violations which occur must be alleged to
represent a duration comparable to the frequency of monitoring...
e.g., monthly monitoring = 30 days.
As with all parameters, permittees who violate the limit
always have the option of conducting more frequent monitoring if
they doubt that the results are truly representative of the entire
period of time.
-110-

-------
2. TRACKING COMPLIANCE AND
DETERMINING VIOLATIONS:
SELF-MONITORING REPORTS
DMR/QA PROGRAM
INSPECTIONS
CITIZEN COMPLAINTS
SELF MONITORING REPORTS:
-	DISCHARGE MONITORING REPORTS
-	Sample Type (required and actual)
-	Sample Frequency (required and actual)
-	Sample Location (required)
-	Analytical Results (limit and actual)
-	SIGNATURE CERTIFYING ACCURACY
-	PROGRESS REPORTS
-	Scheduled milestone events
QUALITY ASSURANCE OF DMR DATA:
QA is an important aspect of whole-effluent toxicity control.
Many Regions and States are requiring complete reports on whole-
effluent toxicity tests as specified in the protocols. These reports
are then compared to QA checklists to make sure that the tests
fall within the parameters of the protocol. Any deviation from
these bounds (temperature, D.O., etc.) provide the basis for
requiring re-tests, or require more technical review (i.e., ESD
review) to determine whether or not the test was valid.
DMR/QA:
REFERENCE TOXICANT PILOT PROJECT
*	New Jersey
*	North Carolina
-111-

-------
PERMITTEE NAMt/ADOREM ttmhnlr
fmihti A.mm,-//	if UiQrre mil
NAME	
ADDRESS
NATIONAL KM.LUTANT DISCNASOf ELIMINATION SYSTEM {NPt>*S)
DISCHARGE MONITORING REPORT Afff)
		/*>			(17-19)	
form VunJ
o*mma 2040-000$
Appro** *Mpirm 9-JO-*
PERMIT NUMBER


MONITORING
PERIOD


FACILITY
YEAR
MO
DAY

YEAR
MO
DAY

LOCATION FROM



TO



MOTE: Road Instructions botore complsMog this term.

{SU JI)
{2113)
(Mi)

(lb 2Tl
| Hit,
iJOJI)
(32-37)
(J Card Only) QUANTITY OR LOAOINO
(46-53)	(*441)
(< Card Only)
OUALITY OR CONCENTRATION
(
******
****
4.8
UIKLY MM
DELMON
MONTH MM
******
PERCENT
MNTHLY
24/COMI'
SAMPLE
MEASUREMENT
_ to
Pamir
roquiremomt
SAMPLE
MEASUREMENT
PfBUSIT
ftMuinasmr
pamiit
ftEQUIRCMBNT
NAME/TITLE PRINCIPAL EXECUTIVE OFFICER
TYPED OR PRINTED
i ccRTirv unocr pcnalrv or law that i havc personally cmaminco
A NO AM F AMI IAR WITH THE lf#OM*AT)OM SUBMITTED MCRflN A NO OASCD
ON MV tNOumv or TmOSC INDIVIDUALS iMMCCNATFIV RtSRONSlOLC fOR
OfTT AIN»*G THf INFORMATION I SClKVC THF SUBMITTED INFORMATION
IS TRUC ACCURATC ANO COMPLtTl I AM A*AR{ THAT THf Rf ARC SlG
NtftCANT PCNAl TICS FOR SUBMITTING FALSI l»f Off MA TON INCIUOING
THf POS^ieniTT Of *INf ANO IMPRISONMCNT Sit 10 USC \ 1001 ANO
33USC 4 <319 iPrnalltr» under thrm* ttmiulru m«v	/mr> U(> IIihumi
artd luaiMM imprtmtnrttrtti tj tulu-rrn 4 muntMm and » \i
SIGNATURE OF PRINCIPAL EXECUTIVE
OFFICER OR AUTHOR IZEO AGENT
TELEPHONE
DATE
AREA
CODE
COMMENT AND EXPLANATION OF ANY VIOLATIONS (Reference all utluihmtnrs here)
TOXICITY TESTINC REPORTS SHALL BE ATTACHED TO AND SUBMITTED WITH THE DISCHARGE MONITORING REPORT. ALL SAMPLES SHALL BE
TAKEN AFTER THE LAST TREATMENT UNIT PRIOR TO DISC11ARCE INTO THE RIVER. NOEL SHALL BE REPORTED AS PERCENT EFFLUENT.
\ Form 3320-1 (R»». 10-79) previous coition to ok used
UNTIL SUPPLY 10 EXHAUSTED
(REPLACES IPA FORM T-40 WHICH MAY NOT BE USED.)
pmir n>

-------
QOMtlTY CONTROL FACT SHEET FOR SELF-BIOMONITORING
ACUTE/CHRONIC TOXICITY TEST DATA
Permit No. ftK OOOOOQI
Facility Nana	AQCOftP.
Facility Location AftWfPUJN
Laboratory/Investigator 		
Permit Requirementsi	_ 	
	 foUWMft if&X	WJ&AH1ED-
Sampling Location fMVK	Type of Sample 1A ttOOff CDHP.
Limit _!i3e		Test Duration 7 PW C°* 3 broods}
Type of Test CgfllPWPMNlA PUftA,	Test Organism Age 2-2.4' ftU. UtttkiN
ttfom s-Q&c-tiei&mi	4 ***& e? era* ptosr
Test Resultsi reftop GlOOl.O)
LCS0/EC50/^OEl)		95% Confidence Interval
Quality Control Summary:
Date of Sample: 	 Dates of Test: IftJrtlHH 72fmft.t\ON
y ^	^ lfS.
Control Mortality: a %	Control Mean iQffyi.i^iiftiiiUpM ¦£» \p
#"*** 26 °"
Temperature maintained within +#»C of test temperature? Yea	 No	
Dissolved oxygen levels always greater than 40% saturation?
YeS	 N°		£ l/i5ml *olu&bn
Loading factor for all exposure chambers less than or equal to
maximum allowed for the test type and temperature? Yes	 No	
Do the test results indicate a direct relationship between effluent
concentration and response of the test organism (i.e., more deaths
occur at the highest effluent concentrations)? Yes	 No	
-113-

-------
INSPECTIONS:
Inspections are conducted to:
*	Verify permittee compliance
*	Support enforcement actions
41 Respond to citizen complaints
*	Support permit development
*	Maintain a regulatory presence
INSPECTIONS FOR WHOLE-EFFLUENT
TOXICITY
*	Part of compliance inspection
*	Focus on QA
*	Regions/States must have the capability of assessing
sample compliance (lab capability in-house or thru
contract)
*	Reserve sampling inspections for permitting and
enforcement priorities
Biomonitoring Inspector Training Module
CITIZEN COMPLAINTS
TRACKING COMPLIANCE through the Permit
Compliance System:
PCS FUNCTIONS:
*	Tracks permit issuance, reissuance, and appeal activities
*	Screens compliance data for effluent, schedule, and
reporting violations
*	Tracks enforcement responses
*	Automates QNCR preparation
*	Automates (some) Strategic Planning and Management
System reporting
*	Provides facility information
*	Tracks inspections
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TABLE 020 COMPLIANCE SCHEDULE EVENTS
(partial TRE list)
Code: Schedule Event:
001CT Submit Toxicity Evaluation Scope
001PA Submit Toxic Reduction Evaluation
002CT Begin Toxicity Evaluation Program
003CT Proposed Toxic Reduction Treat Mod.
01399 Submit Proposed Toxic Reduction Eval
01999 Completion of Toxic Reduction Eval
02199 Toxics Reduction Evaluation Plan
02299 Toxics Reduction Evaluation
10108 1st Bioassay Result
WET PARAMETER CODES
*	Five digits, first one always a "T'
*	Second digit reflects analytical end point
(e.g., LCbo, NOEL, percent mortality, etc.)
0 Third digit reflects type of test (e.g., acute,
chronic), length of test (e.g., 24 hours), and solution renewal
(e.g., static, renewal, or flow-through)
4 Fourth and fifth digits reflect species
In PCS, effluent limits and measurements are tied to a pipe
schedule which specifies the pipe, reporting frequency (e.g.,
monthly DMRs), and limit effective dates. Whole-effluent toxicity
limits that are required on a different reporting frequency, or that
have different limit effective dates, or that change reporting
frequency or limit within the life of the permit require the
establishment of new pipe designators (act like separate pipes).
This can be confusing and time-consuming.
MORAL: Keep differences in reporting frequency and limit
effective dates to a minimum or be prepared for an increase in
mental illness, followed by sore throats (from screaming), a loss of
hair (from pulling it out), and increased head and stomach aches
among the PCS community!
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"T" DATA SHEET
Whole-Effluent Limit Expressed As (circle appropriate):
A = LCm
B = NOEL
C = % effluent causing	effect (e.g., LC3J
D = NOAEL
E = Low flow pass/fail (LF P/F)*
F = Half low flow pass/fail (HLF P/F)*
G = P/F*
H = Chronic Value (CHV)
I = LCgo/P/F*
J = % Mortality at	% effluent
* Pass/Fail Conditions	
Type of Test (circle appropriate):
A = Static 48 hour ACUTE
B = Static 96 hour ACUTE
C = Static 4 day CHRONIC
D = Static 7 day CHRONIC
E - Static 24 hour ACUTE
M = Static Renewal 48 hour ACUTE
N = Static Renewal 96 hour ACUTE
O = Static Renewal 4 day CHRONIC
P = Static Renewal 7 day CHRONIC
W = Flow-thru 48 hour ACUTE
X = Flow-thru 96 hour ACUTE
Y = Flow-thru 4 day CHRONIC
Z = Flow-thru 7 day CHRONIC
Test Species (circle appropriate):
1A = Selenastrum capricornutum
IB = L minor
1C = Champia
3A = Arbacia
3B = Ceriodaphnia
3C = Daphnia magna
3D = Daphnia pulex
3E = Mysidopsis bahia
3F = Oyster embryo
3G = Daphia species
6A = Cyprinodon variega
6B = Menidia
6C = Pimephales promelas
6D = Salmo gair
6E = Lepomis macrochirus
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3. REPORTING AND PLACING THE
VIOLATIONS IN PRIORITY ORDER:
QUARTERLY NONCOMPLIANCE REPORT ©NCR)
-	Effluent limitations - with potential to impact water quality
-	Compliance schedule milestones - 90 days late
-	Reports - 30 days late or incomplete
SIGNIFICANT NONCOMPLIANCE (SNC)
-	Effluent violations - with potential to impact water quality
-	Compliance Schedule Milestones* - 90 days late
-	Submit TRE plan/schedule
-	Initiate TRE
-	Submit final TRE test results/Implementation Plan
-	Start construction
-	End construction
-	Attain final compliance
-	Reports* - 30 days late
•	DMR
•	Final TRE report of progress (indicate permit compliance)
* All milestones and reports required by judicial actions
4. DETERMINING APPROPRIATE RESPONSE:
Initial review of self-monitoring reports leading to the review
of violations by someone responsible for enforcement:
VIOLATION REVIEW ACTION CRITERIA
(magnitude of violation that warrants professional review)
VRAC = 1
Any violation of a Whole-Effluent Toxicity limit must be
evaluated
Any monitoring results in response to "monitor only"
requirements must be evaluated
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HOW DO YOU REVIEW TOXICITY VIOLATIONS?
CONSERVATIVELY!
Remember assumptions made in permit limit
derivation
-	Variability of Effluent
-	Monitoring Frequency
-	Acute to Chronic Ratio
Consider importance of a prompt response
-	Time needed to determine cause/eliminate toxicity
-	Is stream near (or soon may be near) low flow conditions?
Do you have definitive data, or just a
pass/fail result?
WHAT NEXT?
Same first step as with any violation:
GET INFORMATION ON THE VIOLATION
Require More Monitoring if necessary
Remind permittee of violation reporting requirements:
40 CFR Section 122.41 (1)(6&7)
Reporting Requirements. (6) "The permittee shall report any
noncompliance which may endanger health or the environment.
Any information shall be provided orally within 24 hours from the
time the permittee becomes aware of the circumstances. A
written submission shall also be provided within 5 days of the
time the permittee becomes aware of the circumstances. The
written submission shall contain a description qf the
noncompliance and its cause; the period qf noncompliance,
including exact dates and times, and (f the noncompliance has
not been corrected, the anticipated time it is expected to
continue; and steps taken or planned to reduce, eliminate and
prevent reoccurrence qf the noncompliance...(7)...The permittee
shall report all instances of noncompliance not reported under
paragraphs (1)(4), (5), and (6)...at the time monitoring reports are
submitted. These reports shall contain the information listed in
paragraph (1)(6) of this section."
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HOW DO PERMITTEES IDENTIFY THE SOURCE
OF THE TOXICITY - AND ITS TREATABILITY?
GENERALIZED TRE FLOWCHART
Information and Data
Acquisition
Toxicity Identification
Evaluation
Toxicity Treatability
Evaluation
Control Method Selection
and Implementation
HOW DO YOU REQUIRE A TRE?
Mechanisms:
Permit - can require monitoring, establish effluent limitations, and
require any and all phases of a TRE, including construction.
Section 308 Letter - can require monitoring and the first phases
of a TRE (toxicity source and treatability studies, not
construction).
Section 309 Orders, Judicial Decrees - can require monitoring and
all phases of a TRE, including constructions, BUT ONLY IN
RESPONSE TO A PERMIT VIOLATION!
NOTE: Only permits may require a compliance schedule for
construction or other corrective measures in advance of the
effective date of a whole-effluent limit in the permit.
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DETERMINING HOW TO WORD A TRE
SCHEDULE:
Easier if you have some information in advance (require as
much as possible informally)
Make sure you have a FINAL COMPLIANCE DATE (it may be
your last strong-hold)
Remember it is the PERMITTEE'S responsibility to solve their
problem (EPA is providing the protocols and some assistance
thru NETAC, BUT we do not have enough Don Mounts, Fred
Bishops, Bill Peltiers, etc., etc., to solve the nation's problems
MODEL LITIGATION GUIDANCE
*	Anticipated defenses with responses
*	Statutory and Regulatory cites
*	Model complaint alleging WET monitoring and
limit violations
*	Model consent decree requiring a TRE
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STATUTORY AND REGULATORY CITES
Statute:
Section 101(a)
"The objective of this Act is to restore and maintain the
chemical, physical and biological integrity of the Nation's
waters."
Section 101(a)(3)
Declaration of Goals and Policy • "it is the national policy
that the discharge of toxic pollutants in toxic amounts be
prohibited;"
Section 301(a)
"Except as in compliance with this section and sections
302,306,307, 318, 402, and 404 of this Act, the discharge
of any pollutant by any person shall be unlawful." - see
Section 502
Section 301(b)(1)(C)
'In order to carry out the objective of this Act there shall
be achieved not later than July 1, 1977, any more stringent
limitation, including those necessary to meet water quality
standards...
Section 302(a)
provides the authority to establish water quality-based
effluent limitations on discharges that interfere with the
attainment or maintenance of that water quality which shall
assure protection of public health, public water supplies,
and the protection and propagation of a balanced population
of shellfish, fish and wildlife.
Section 303(c)(2)(B)
authorizes the adoption of numeric water quality criteria
that are based upon biological monitoring or assessment
methods and the use of effluent limitations or other permit
conditions based on or involving biological monitoring or
assessment methods or previously adopted numeric criteria.
"Nothing in this section shall be construed to limit or delay
the use of effluent limitations or other permit conditions
based on or involving biological monitoring or assessment
methods..."
Section 304(a)(8)
requires EPA to develop and publish information on
methods for establishing and measuring water quality
criteria for toxic pollutants including biological monitoring
and assessment methods.
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Section 308(a)
authorizes the installation, use and maintenance of
biological monitoring methods by point sources, where
appropriate, for the development of effluent limitations or
the determination of compliance with such limitations,
prohibitions, or effluent standards.
Section 402
authorizes issuance of a permit for the discharge of any
pollutant, or combinations of pollutants (See Section 502),
upon the condition that the discharge meet all applicable
requirements and provisions of the CWA.
Section 502
"effluent limitation" is defined as 'any restriction...on
quantities, rates, and concentrations of chemical, physical,
biological, and other constituents which are discharged..."
"pollutant" is defined as '...industrial, municipal, and
agricultural waste discharged into water.'
"toxic pollutant" is defined as 'those pollutants, or
combinations of pollutants, including disease-causing agents,
which after discharge and upon exposure, ingestion,
inhalation or assimilation into any organism, either directly
from the environment or indirectly by ingestion through
food chains, will... cause death, disease, behavioral
abnormalities, cancer, genetic mutations, physiological
malfunctions (including malfunctions in reproduction) or
physical deformations, in such organisms or their offspring.
"biological monitoring" is defined as 'the determination of
the effects on aquatic life, including accumulation of
pollutants in tissue, in receiving waters due to the
discharge of pollutants (A) by techniques and procedures,
including sampling of organisms representative of
appropriate levels of the food chain appropriate to the
volume and the physical, chemical, and biological
characteristics of the effluent, and (B) at appropriate
frequencies and locations."
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Regulation - 40 CFR:
Section 122.2
'Whole effluent toxicity means the aggregate toxic effect of
an effluent measured directly by a toxicity test.
Section 122.44(d)
'In addition...each NPDES permit shall include conditions
meeting the following requirements when applicable...Water
quality standards and State requirements: any requirements
in addition to or more stringent than promulgated effluent
limitations guidelines or standards...necessary to:
(1) achieve water quality standards established under section
303 of the CWA, including State narrative criteria for water
quality
(i) Limitations must control all pollutants or pollutant
parameters...which...may be discharged at a level
which will cause, have the reasonable potential to
cause, or contribute to an excursion above any State
water quality standard, including State nan-ative
criteria for water quality...
(iv)	(Numeric criterion for whole effluent toxicity)
(v)	'...has the reasonable potential to cause, or
contributes to an in-stream excursion above a
narrative criterion..., the permit must contain effluent
limits for whole effluent toxicity...[except] where
chemical-specific limits for the effluent are sufficient
to attain and maintain applicable numeric and
narrative State water quality standards.'
COURT DECISION:
NRDC v. EPA. 859 F.2d 156 (D.C. Cir. 1988)
The court found that "while 'toxicity' appears to be an
attribute of pollutants rather than a pollutant itself, we see no
reason why this should preclude the agency from using it as a
measure to regulate effluents that are pollutants. Under Section
502(6) of the Act, 33 U.S.C. Section 1362(6), the term 'pollutant'
includes 'industrial, municipal, and agricultural waste.' Any
discharge to which a toxicity limit could be applied would seem to
fall within this broad definition."
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ENFORCEMENT CASES
NRDC v. EPA. 859 F.2d 156 (D.C. Cir. 1988)
The court upheld EPA regulations which authorize the use
of effluent limits framed in terms of toxicity.
Reynolds Metals Co. v. EPA. 760 F.2d 549 (4th Cir. 1985);
Weyerhaeuser Co. v. Costle. 590 F.2d 1011 (D.C. Cir. 1978);
C&H Sugar Co. v. EPA. 553 F.2d 280 (2nd Cir. 1977);
FMC Corp. v. Train. 539 F.2d 973 (4th Cir. 1976); and
BASF Wyandotte y. Costle. 598 F.2d 651 (1st Cir. 1979)
The courts upheld other water characteristics (BOD, TTO,
TSS, COD) as permit limits.
Champion International Corp. y. EPA. 648 F. Supp. 1398 (W.D.N.C.
1987)
The court upheld EPAs authority to object to permits which
do not contain conditions adequate to achieve approved state
water quality standards.
Trustees for Alaska v. EPA. 749 F.2d 549, 557 (4th Cir. 1984)
The court found that EPA as permit writer is required to
establish whatever permit limits are necessary to achieve water
quality standards.
API v. EPA. 787 F.2d 978 (5th Cir. 1986)
The court upheld EPAs use of a 96-hour LCM as the most
widely accepted benchmark for toxicity evaluations by EPA and
sustained EPAs choice of the test for limiting effluent "mud"
(drilling fluid) toxicity.
BASF Wyandotte v. Costle. 598 F.2d 647-50. 655 (1st Cir. 1979);
Citizens to Preserve Overton Park v. Volpe. 401 U.S. 402, 416,
91 S.Ct. 814, 824, 28 L.Ed.2d 136 (1971);
Permian Basin Area Rate Cases. 390 U.S. 747, 810-11, 88 S.Ct.
1344, 1382-83, 20 L.Ed.2d 312 (1968); and
Baltimore Gas & Electric Co. v. Natural Resources Defense
Council. 462 U.S. 87, 103 (1983)
The courts deferred to the Agency's judgement in the
settlement of technical issues.
Given the on-going nature of enforcement of water quality-
based permit requirements, it is recommended that updated
information be researched by the reader through available means
such as the use of LEXIS.
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Section 5
Basic Permitting Principles
Introduction to Toxicity Reduction Evaluations (TRE)
TREs in Permitting and Enforcing Process
TRE Available Guidance
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PERMITTING PRINCIPLES AND
EXAMPLE PERMIT LANGUAGE
FOR TREs
BASIC PERMITTING PRINCIPLES
AND
EXAMPLE PERMIT LANGUAGE
FOR
WET and TREs
PRINCIPLE NUMBER ONE
PERMITS MUST BE
PROTECTIVE OF WATER
QUALITY


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AT A MINIMUM, ALL MAJOR PERMITS AND
MINORS OF CONCERN SHOULD BE
EVALUATED FOR POTENTIAL OR KNOWN
TOXICITY
FINAL WHOLE EFFLUENT TOXICITY LIMITS
MUST BE INCLUDED IN PERMITS WHERE
NECESSARY TO ENSURE THAT STATE
WATER QUALITY STANDARDS ARE MET
PRINCIPLE NUMBER TWO
PERMITS MUST BE
WRITTEN TO AVOW
AMBIGUITY AND ENSURE
EN FOR CEABILITY
PERMIT CONDITION NUMBER
ONE
PERMIT LIMITS IN PART ONE
OF PERMIT EFFECTIVE IMMEDIATELY
OR AT A SPECIFIED DELAYED
DATE
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EXAMPLE PERMIT LANGUAGE
FOR EFFLUENT TOXICITY LIMITS
Part 1A. Final Eftluent Limitations and Monitoring Requirements
During the period beginning on the effective date
of this permit and lasting until the expiration date,
the permittee is authorized to discharge in
accordance with the following limitations and
monitoring requirements from the following
outfall(s):	
Effluent Characteristic
Discharge Limitation
Monnonng Frequency
Reporting
code/units
Parameter
Daily
Maximurr
Monthly
Average
Measurement
Frequency
Sample
Type
61426/TUc Toxicity 5.9	1.2	x/mofflfi composite
PERMIT CONDITION NUMBER
TWO
REPORTING REQUIREMENTS TO INCREASE
MONITORING IN THE EVENT OF EFFLUENT
VIOLATION, AND LEADING EITHER TO
AN ENFORCEMENT ACTION, TRE OR
RETURN TO NORMAL MONITORING
DOES NOT VERIFY THE ORIGINAL VIOLATION
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EXAMPLE PERMIT LANGUAGE
FOR REPORTING REQUIREMENTS
Pari 1B. Reporting Requirements
1. Toxicity Limitations
Where any one monitoring event shows a violation of the limits
in Part 1A of this permit, the permittee shall be considered in
violation of this permit and shall increase the frequency of
toxicity testing to once per week and submit the data within
x days to the permitting authority The permitting authority
will determine whether enforcement action will be initiated, or
whether the permittee must implement the requirements of
Part IIIA of this permit or return to the monitoring requirements
in Part 1A.
The permittee shall use the testing and data assessment procedure
described in Part IIIB of this permit.
PERMIT CONDITION NUMBER
THREE
TOXICITY TESTING SPECIES AND
PROTOCOLS SHOULD BE ACCURATELY
REFERENCED/CITED IN THE PERMIT
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PRINCIPLE NUMBER THREE
WHERE NOT IN COMPLIANCE
WITH A WET LIMIT,
PERMITTEES MUST BE
COMPELLED TO COME INTO
COMPLIANCE WITH THE
LIMIT AS SOON AS POSSIBLE
ALL COMPLIANCE DATES SHOULD BE
SPECIFIED
CORRECTIVE ACTIONS (TREs) CANNOT
BE DELAYED PENDING EPA OR STATE
APPROVAL OF THE PLAN. UNLESS
STATE REGULATIONS REQUIRE
PRIOR APPROVAL
TRE PRELIMINARY SCHEDULE
SET IN PERMIT WHERE TOXICITY IS
KNOWN, OR BY RE-OPENING THE
PERMIT OR ISSUING AND ENFORCEMENT
ORDER (SECTION 308 OR 309) WHERE
TOXICITY IS FOUND SUBSEQUENT
TO PERMIT ISSUANCE
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EXAMPLE PERMIT LANGUAGE
FOR TRE SCHEDULES
Part MIA. Special Conditions: Toxicity Reduction Evaluation
The Discharger shall demonstrate that effluent toxicity-based permit
limits described in Part IA of this permit are being attained and
maintained through the application of all reasonable treatment and/or
source control measures. Upon identifying noncompliance with those
limits following the conditions of Part IC1, the Discharger shall initiate
a TRE according to the following schedule:
IaSk	Deadline
1. Take all reasonable measures necessary
to immediately reduce toxicity, where
source is known
Within 24 hours
2.	Where source of toxicity is known, submit
a plan and schedule to attained continued
compliance with effluent toxicity-based permit
limitations in Part IA, if immediate compliance
is not attained
3.	Where source of toxicity is unknown and
toxicity cannot be immediately controlled
through operational changes, submit a TRE
study plan detailing the toxicity reduction
procedures to be employed. EPA's Toxicity
Reduction Evaluation Procedures; Phases 1,2 and
3 (EPA 600/3-88-034,035 and 036) and TRE
Protocol for POTWs (EPA600/8-88-00)
shall be the basis for this plan.
4.	Initiate TRE plan
5.	Comply with approved TRE schedule
6.	Submit results of TRE; include a summary of
findings, corrective actions required, and data generated
7.	Implement TRE controls as described In final report
8.	Complete TRE implementation to meet
permit limits and oonditions
Within 30 days
Within 45 days
Within 45 days
Immediately upon
approval
Per approved schedule
On due date of final reportj
per approved schedule
Per approved schedule,
but in no case later than
later than x months from
initial noncompliance
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TOXICITY REDUCTION EVALUATION
(TRE)
A SITE-SPECIFIC STUDY CONDUCTED IN
A STEP-WISE PROCESS TO
NARROW THE SEARCH FOR
EFFECTIVE CONTROL MEASURES
FOR EFFLUENT TOXICITY

Figure 1 - Generalized TRE Flowchart
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WHAT PROMPTS A
TOXICITY REDUCTION
EVALUATION?
SCENARIO #1
TOXICITY FOUND PRIOR
TO PERMIT ISSUANCE
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SCENARIO #1
® TRE REQUIREMENT IN PERMIT
© SOME DATA. SO SPECIFIC SCHEDULE
• LIMIT IN PERMIT PRIOR TO REQUIRING
CONSTRUCTION OR SOURCE CONTROL
EXAMPLE PERMIT LRN6URGE
FOR TRE SCHEDULES
Part IIIR. Special Conditions: Toxicity Reduction Evaluation
The Discharger shall initiate a TRE according to the following schedule
Task
Deadline
1 Submit TRE study plan
detailing the toxicity reduction
evaluation procedures to be
employed EPAs Toxicity Reduction
Evaluation Procedures: Phases 1. 2
and 3 (EPA 600/3-88-034.035 and
0361 and TRE Protocol for Municipal
Wastewater Treatment Plants (EPA
600/8-88-621 shall be the basis for
this plan
Within 45 days of
permit Issuance
2 Inmate TRE
3. Submit TRE progress reports
4 Submit results of TRE
5 Implement TRE controls as
described in final report
Within 45 days of
permit issuance
By the 15th day of each
calendar quarter
Within 10 months of
permit issuance
Within 16 months of
permit issuance
6 Complete tojdcity control
implementation and meet
permit limits and conditions
Within 2 years of
permit issuance
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^ SCENARIO #2A
TOXICITY TESTING WITH A
TRE TRIGGER
SCENARIO #2A
• TRE TRIGGER IN PERMIT
• NO SPECIFIC DATA, SO GENERAL TRE
SCHEDULE
• MUST REOPEN PERMIT TO SET LIMIT
AND REQUIRE CONSTRUCTION,
SOURCE CONTROL. OR OTHER
TOXICITY CONTROL OPTIONS
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EXAMPLE PERMIT LANGUAGE
FOR TRE SCHEDULES
Part ll IA Special Conditions Toxicity Reduction Evaluation
The Discharge' shall demonstrate mil affluent loudly-based permit
limits DttcMd in Part IA ot this permit a'* Ming attained and
maintained through the application o> all reasonable treatment tnti/or
source control measures Upon identifying noncompliance with those
limns following the conditions ot Pan IC1. the Discharger shall initiate
a TRE aocon*ng 10 the following schedule
T«sfc	Da«r*n>
t Take all reasonable measuros necessary
to immediately reduce toiejry, where
lourco is known	Within 24 hours
2	Where source ot toitoty is known, submit
a plan and schedule to attaned oonwiued
compliance arttn etttuent to>citybased permit
bmltatens m Pan IA, H Immediate oompkance
la not attalnod
3	Where source ot k»oty is unknown and
toocity cannot be Immediately controlled
through operational changes, submit • TRE
study plan (totalling ttte totdty reduction
procedures to be employed EPAs Ternary
Reduction Evaluaton Procedures Phases t.2 and
3	(EPA S0O/3'S8-034.0J5 and 036) and TRE
Protocol tor POTWs (EPASOO'S SS OOI
tihal b» the bant tor tha plan
4	Initiate TRE plan
5. Comply with improved TRE schedule
4 Submit results ot TRE. nckjdt a summary o'
findings, corrective actions requirod. and data generated
7 Implement TRE controls as described n final repon
0. Complete TRE Implementation to meet
permit bmita and oondftens
Within 30 days
Within 45 days
Wittifi 45 days
tmmeckately upon
approval
Pot approved schedule
On due date ot fmai report
per approved schedule
Per approved schedule,
but in no case later than
Wet than > months from
Initial noncompliance
rr SCENARIO #2B
TOXICITY TESTING WITH
NO TRE TRIGGER
OR
PERMITTING AUTHORITY
FINDS TOXICITY
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SCENARIO #2B
1.	ISSUE SECTION 308 LETTER TO
REQUIRE ACCELERATED TESTING
2.	IF ADDITIONAL DATA SHOWS TOXICITY,
ISSUE SECTION 308 LETTER TO REQUIRE
TRE
REOPEN PERMIT TO ADD LOOT AND/OR
REQUIRE TRE
® SOME DATA, SO SPECIFIC SCHEDULE
• MUST REOPEN PERMIT TO SET LIMIT
AND REQUIRE CONSTRUCTION OR
SOURCE CONTROL. OR OTHER
TOXICITY CONTROL OPTIONS
KF SCENARIO #3A
WHOLE EFFLUENT TOXICITY
LIMIT WITH A TRE TRIGGER
OR
SCENARIO #2B
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SCENARIO #3A
•	NO SPECIFIC DATA, SO GENERAL TRE
SCHEDULE
•	MAT WANT TO ISSUE SECTION 308
LETTER TO REQUIRE ACCELERATED
TESTING
•	DO NOT NEED TO REOPEN PERMIT TO
SET LIMIT/REQUIRE CONSTRUCTION.
SOURCE CONTROL. OR OTHER
TOXICITY CONTROL OPTIONS
• MAT WANT TO REOPEN PERMIT TO
MODIFT SCHEDULE
SCENARIO #3B
WHOLE EFFLUENT TOXICITY
LIMIT WITH NO TRE TRIGGER
SCENARIO #3B
•	ISSUE SECTION 308 LETTER REQUIRING
ACCELERATED MONITORING
OR
•	ISSUE SECTION 309 ORDER REQUIRING
ADDITIONAL MONITORING AND/OR TRE
• SOME DATA, SO SPECIFIC TRE SCHEDULE
• DO NOT NEED TO REOPEN PERMIT TO
SET LIMIT/REQUIRE CONSTRUCTION.
SOURCE CONTROL. OR OTHER
TOXICITY CONTROL OPTIONS
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OTHER CONSIDERATIONS
A SUCCESSFUL THE SHOULD RESULT IN
COMPLIANCE WITH THE PERMIT
TOXICITY MAT BE ELIMINATED THROUGH
SIMPLE OfltM OR HOUSEKEEPING
IMPROVEMENTS
TOXICITY MAT MYSTERIOUSLY DISAPPEAR
BUT THE TRE IS NOT OVER YET
-EXISTING DATA EVALUATION
-CONTINUED FOLLOW-UP
MONITORING

BJP IN SUMMARY. . .
TREs
ARB TRIGGERED BY "UNACCEPTABLE .
TOXICITY1'
CAN BE REQUIRED BT
PERMIT [Q
SECTION 308 LETTER
SECTION 309 ORDER (OR
JUDICIAL DECREES)
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AVAILABLE GUIDANCE
•	METHODS FOR AQUATIC TOXICITY
IDENTIFICATION EVALUATIONS
-	PHASE I—TOXICITY CHARACTERIZATION
PROCEDURES, EPA 600/3-88/034
-	PHASE II—TOXICITY IDENTIFICATION
PROCEDURES, EPA 600/3-88/035
-	PHASE III—TOXICITY CONFIRMATION
PROCEDURES, EPA 600/3-88/036
•	GENERALIZED METHODOLOGY FOR CONDUCTING
INDUSTRIAL TOXICITY REDUCTION EVALUATIONS,
EPA 600/2-88/070
•	TOXICITY REDUCTION EVALUATION PROTOCOL
FOR MUNICIPAL WASTEWATER TREATMENT
PLANTS, EPA 600/2-88/062
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Figure i 2 Tonciry BtOoctton Evauiauon (TRE) Mow cnan.
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Yll
Convention*! Pollutant
Trttiabilitv Tam
Toncitv Identification
Evaluation (F^jur# 4-1)
vaa
Figure 1-1. TRC flew diagram for municipal waatawatar trsatmant plant.
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Section 6
TRE Municipal and Industrial Protocols
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MUNICIPAL PROTOCOL
TOXICITY REDUCTION EVALUATION
PROTOCOL FOR MUNICIPAL
WASTEWATER TREATMENT PLANTS
TRE REQUIREMENT
•	Triggered by evidence of unacceptable
effluent toxicity
•	Usually a TRE plan and schedule must be
submitted
•	Continues until acceptable effluent toxicity
is achieved
TOXICITY REDUCTION EVALUATION
•	Identify the constituents causing effluent
toxicity
•	Locate the sources of effluent toxicants/toxicity
•	Evaluate the feasibility and effectiveness of
toxicity control options
-147-

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MUNICIPAL TRE PROTOCOL
•	Development and review of a TRE plan
•	Selection of appropriate steps in a TRE
•	Evaluation and interpretation of the data
•	Selection and implementation of control
options
LIMITATIONS OF THE PROTOCOL
•	Addresses Methods for Reduction in Whole
Effluent Toxicity
•	Limited Case Studies
COMPONENTS OF THE MUNICIPAL TRE PROTOCOL
Information and Data Acquisition
POTW Performance Evaluation
Toxicity Identification Evaluation
Toxicity Source Evaluations (Tiers I and II)
POTW In-Plant Control Evaluation
Toxicity Control Selection and Imolementation
POTW OPERATIONS AND PERFORMANCE DATA
NPDES Discharge Monitoring Reports
POTW Design Criteria
Process Control Data
Treatment Interferences
Process Sidestream Discharges
Wastewater Bypasses
-148-

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PRETREATMENT PROGRAM INFORMATION
POTW Effluent and Influent Toxicity/Toxics Data
POTW Sludge Toxics Data
Industrial Waste Survey Information
Annual Pretreatment Program Reports
Local Limits Compliance Reports
POTW PERFORMANCE EVALUATION
•	Evaluate major unit treatment processes
(CCP Approach)
•	Identify deficiencies that may contribute to
effluent toxicity
•	Determine in-plant sources of effluent toxicants
(e.g., chlorination, bypasses)
POTW PERFORMANCE EVALUATION
A limited TIE Phase I can be conducted to:
•	Indicate in-plant toxicants such as chlorine
and suspended solids
•	Provide information to set up treatability tests
CONVENTIONAL TREATABILITY TESTS
Recommended for Improvements in Conventional
Pollutant Treatment
Can Identify Modifications in Conventional
Treatment That Also Reduce Toxicity
-149-

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CONSIDERATIONS IN TIE TESTING AT POTWS
Characterize effluent toxicant variability over time
Utilize pretreatment program data to support TIE
Can initiate treatability tests based on Phase I
results
RESULTS OF TIE
•	Specific toxicants are identified
•	One fraction is consistently toxic
•	Variable fraction toxicity
PURPOSE OF TOXICITY SOURCE EVALUATION (TSE)
Determine Sources of Effluent Toxicants/Toxicity
Determine Feasibility of Pretreatment Control
TIER I TSE - SAMPLING DECISIONS
Sewer Line Sampling:
•	TIE and pretreatment program data
are limited
•	POTW has a large number of lUs
Point Discharge Sampling:
•	TIE and pretreatment program data
attribute toxicants to lUs
•	Number of lUs is manageable
-150-

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TSE TIER I APPROACHES
Chemical-Specific Tracking
Refractory Toxicity Assessment
CHEMICAL-SPECIFIC TSE REQUIREMENTS
Pretreatment Program Data to Indicate Sources
Knowledge of Sewer Discharge Characteristics
Accurate Analytical And Flow Data
TSE - REFRACTORY TOXICITY ASSESSMENT
A simulation of the POTW treatment system
which utilizes toxicity tests to estimate the
amount of refractory toxicity in sewer
wastewaters.
RTA SAMPLE COLLECTION, CHARACTERIZATION
	AND PREPARATION	
24-Hour Flow Composites
Analyze for COD, TKN, TP, TDS and pH
Adjust BODs:N:P Ratio to 100:5:1
Adjust pH
-151-

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EXAMPLE RESULTS FOR TIER I RTA
LC50 (%	Effluent)
Sample/	Sample/	Potential
Synthetic	Primary	Primary	Toxicity
Source Wastewater	Effluent	Effluent	Source
A 35 38	70	YES
B 22 77	72	NO
C 21 12	85	YES
TIER II - TOXICITY SOURCE EVALUATION
Confirm Sources of Refractory Toxicity
Identified In Tier I
Determine Potential for Biological Treatment
Inhibition (optional)
Characterize Refractory Toxicity Using TIE
Phase I Tests (optional)
EXAMPLE RESULTS FOR TIER II RTA
Sample Dilution
(Times Percent Flow in POTW Influent)

10x
5x
2x
Batch Effluent LC 50
10
30
50
Batch Effluent Toxic
10
3.3
2
Units (TU)
Sum of TUs = 15.3
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RELATIVE TOXICITY LOADING CALCULATION
Relative Score =
Sum of TUs x Sewer Discharge Flow Rate
Where Sum of TUs = 15.3
Flow Rate = 1 mgd
Relative Score = 15.3 TU x 1 mgd = 15.3
TSE TIER II - PRETREATMENT CONTROL EVALUATION
Approaches to Local Limits Development
•	Allowable headworks loading
•	Industrial User management
•	Case by case permitting
Equitable Cost Recovery
SELECTION OF OPTIONS FOR EVALUATION
Review PPE data to determine:
•	Space and equipment
•	Operational control
Review TIE data to determine:
•	Types of toxicants amenable to
treatment
•	Treatability test design
-153-

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TOXICITY TREATABILITY TESTS
Activated Sludge
Coagulation and Precipitation
Sedimentation
Granular Media Filtration
Activated Carbon
EVALUATION OF TOXICITY CONTROL OPTIONS
Selection based on results of:
•	PPE
•	TIE
•	TSE Tier I - Chemical Specific Testing
•	TSE Tiers I and II - Refractory
Toxicity Assessment
•	POTW Treatability Testing
POTW TECHNOLOGIES FOR CATEGORIES OF POLLUTANTS
Biodegradable
Organic
Compounds and
Ammonia
Biological Process
Control
Nutrient
Addition
Non-Biodegradable
Organic
Compounds
Coagulation/
Precipitation
Filtration
Activated
Carbon
-154-

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POTW TECHNOLOGIES FOR CATEGORIES OF POLLUTANTS
Organic
Compounds
Volatile
Heavy Metals
and Cationic
Compounds
Biological Process
Control
pH Adjustment
Aeration
Coagulation/
Precipitation
Filtration
COMPARISON OF SELECTION CRITERIA FOR
TOXICITY CONTROL OPTIONS
Ability to achieve effluent toxicity limits
Ability to comply with other permits
Capital and O&M Costs
Ease of Implementation
Reliability
Environmental Impact
TOXICITY CONTROL IMPLEMENTATION
Toxics Control Implementation Plan
Follow-up Monitoring
Selection Criteria
Alternative
ABC
-155-

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Case Study
Toxicity Reduction Evaluation
At The
Linden Roselle Wastewater Treatment Plant
Purpose Of The Case Study
To Refine Procedures For Conducting TREs
With Emphasis On Methods For Tracing
The Sources Of Acute Effluent Toxicity
TRE Elements
WWTP Performance Evaluation
Pretreatment Program Evaluation
Toxicity Identification Evaluation
Toxicity Source Evaluation
Pretreatment Program Evaluation
Average Influent Ammonia = 80 mg/1
Average Effluent Ammonia = 76 mg/1
Variability In Influent Characteristics Related To Variable Industry
Production Schedules
-156-

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WWTP Performance Evaluation
Pretreatment Agreement Reduced Influent BOD5
And TSS Loadings
Lower Influent Loadings Resulted In Improved Treatment
No Net Ammonia Removal Due To Lack Of Nitrification Capability
140 -
_ 130 -
1986	1987	1988
AVERAGE MONTHLY VALUES
Average monthly Influent ammonia concentrations>
Linden Roselle WWTP.
Species Correlation Results
LC50 (Percent Effluent)
Sampling Mysidopsis Daphnia Ceriodaphnia Pimephales
Date	bahia	pulex	dubia	promelas
6/11/89 to 22.1	19.4	54.7	48.2
6/12/89
6/20/89 to 15.8	11.6	17.3	34.9
6/21/89
6/26/89 to 5.2	14.0	17.3	23.3
6/27/89
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TIE Phase I Results
TIE Treatment
A
7/10/89
Ceriodaphnia LC50
B
7/25/89
C
8/23/89
Baseline Toxicity
pH Adjustment
pH 11
Filtration
pH 11
pH 3
Aeration
Original pH
pH 11
C^g SPE Column
Original pH
pH 11
pH 3
75.0
84.5
>100
>100
>100
86.6
NR
>100
>100
39.6
>100
>100
NR
NR
65.0
>100
>100
>100
31.1
>100
>100
NR
NR
>100
42.7
41.6
>100
TIE Phase I Summary
Lower Toxicity For One Sample With Low Ammonia Concentration
Graduated pH Test Suggested Ammonia Or Ammonia-Type
Compounds
Ammonia Reduction In Cj8 SPE Column Treatment
Toxicity Source Evaluation
Bench-scale, Batch Simulation Of The WWTP Activated Sludge
Treatment Process to Determine The Refractory Toxicity Of Sewer
Wastewaters
-158-

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Refractory Toxicity Assessment
Selection Of Key Manhole Sampling Locations
Conventional Pollutant Analyses To Determine BOD:N:P Ratio for
RTA Tests
Develop Sampling Schedule Based On Industry Production Schedules
Sewer sampling locations, Linden Roselle TRE Case Study.
-159-

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Tier I - Toxicity Source Evaluation
Ceriodaphnia LC50
Wastewater
Source
Test
Date
Wastewater In
Primary Effluent
Primary
Effluent
Manhole 7
7/25/89
8/1/89
>100
<10.0
91.3
22.3
Manhole 8
7/27/89
8/1/89
31.1
<10.0
45.8
31.1
Manhole 9
7/24/89
7/31/89
58.6
35.4
35.4
35.4
Tier I - Toxicity Source Evaluation
3 Of The 7 Manholes Were Identified As Possible Sources Of Acute
Refractory Toxicity
2 Of The 3 Toxic Manholes Had Total Ammonia Concentrations Of
200 To 300 mg/1
Several Industries On Manholes 7 And 8 Selected For Further Testing
Sample Number
188851 KM7 TU (FW = 5)	PE TU
-160-

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Sample Number
ESS% KM8 TU (FW - 5)	PE TU
1	2	3	4	5
Sample Number
B88SJ Industry A-7 (2X)	PE Batch Effluent
-161-

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Samplo Number
188881 Industry A-7 (IX)	PE Batch Effluent
C—8	C-8	D—8	D—8	D-8	E-8	E-8
Industry Sample
E8881 Industry TU (5X)	PE Til
-162-

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Summary Of Toxicity Source Evaluation
3 Of The 7 Manholes Were Identified As Possible Sources Of Toxicity
3 Of 6 Indirect Dischargers Were Identified As Possible Sources Of
Toxicity
1 Of The Indirect Dischargers Had Total Ammonia Concentrations
Of 500 To 900 mg/1
Conclusions
RTA Test Was An Effective Tool For Tracing The Sources Of
Refractory Toxicity
TIE Phase I Tests Indicated Ammonia Or Ammonia-Type
Compounds and Non-Polar Organics
Refractory Acute Toxicity Was Associated With Sources of Ammonia
Recommendations
TIE Phase II Analyses For Ammonia And Ammonia-Type
Compounds
Additional TSE Analyses Using A Chemical-Specific And/Or Toxicity
Tracking Approach
Evaluation Of The Nitrification Capability Of The WWTP
-163-

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CASE STUDY
MOUNT AIRY, NORTH CAROLINA
TOXICITY REDUCTION EVALUATION
MT. AIRY TOXICITY REDUCTION EVALUATION
TECHNICAL APPROACH
•	CHEMICAL MEASUREMENTS
•	MOCK EFFLUENTS
•	TIE PHASE I TESTS
•	SOURCE EVALUATION
•	LOCAL PRETREATMENT LIMITATIONS
-164-

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MODIFIED TIE PHASE I
CATION RESIN - METALS
ORGANIC RESIN " ORGANICS
CHEMICAL MEASUREMENT
•	FOCUSED ON ALKYL PHENOLS,
METALS, SOLVENTS
•	COMPARISON OF CHEMICAL DATA
TO LITERATURE TO INDICATE TOXICANTS
-165-

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SOURCE EVALUATION
ALKYL PHENOLS -
COPPER
ZINC
SOLVENTS
TEXTILE SURFACTANTS
DYE COMPONENTS
SODIUM HYDROSULFITE
TEXTILE SCOURING
PRETREATMENT CONTROL
•	LINEAR ALCOHOL ETHOXYLATES IN LIEU OF
ALKYL PHENOL ETHOXYLATES
•	CHEMICAL USAGE OPTIMIZATION
•	REDUCED APPLICATION OF METAL-BASED DYES
•	ZINC-FREE HYDROSULFITE
-166-

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CASE STUDY
TOXICITY REDUCTION EVALUATION
AT THE
PATAPSCO WASTE WATER
TREATMENT PLANT
Baltimore, Maryland
PURPOSE OF TRE CASE STUDY #1
Develop and validate procedures
for municipal TREs with emphasis on
evaluating methods for tracing
effluent toxicity to its sources.
-167-

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WHY PATAPSCO WAS CHOSEN FOR A CASE STUDY
•	Effluent Toxicity
•	Treatment Performance Problems Related
To Toxicity
•	Experience in Toxicity Monitoring/Existing
Data Base
•	Proximity to the Chesapeake Bay Estuary
OBJECTIVES OF THE TRE
•	Evaluate Operations and Performance
•	Identify Effluent Toxicants
•	Trace Toxicants and/or Toxicity
•	Evaluate, Select and Implement
Controls
-168-

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PATAPSCO TRE - CASE STUDY SCHEDULE
MONTHS
0 1 2 3 4 5 6 7 8 9 10 11 12131415161718
TOXICITY TESTING
PERFORMANCE REVIEW
TIE (PHASE I)
SOURCE EVALUATION
FINAL REPORT
AQUATIC TOXICITY TESTS
TEST
ENDPOINT
7-day Ceriodaphnia dubia 48-hour LC 50
7-day ChV
96-hour Mysidopsis bahia 96-hour LC
50
MICROTOX
TM
5-minute EC 50
-169-

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ACUTE TOXICITY OF PRIMARY EFFLUENT
ACUTE TOXICITY OF PRIMARY EFFLUENT
MEAN
lc50/ec50 (S.D.)	N
Ceriodaphnia dubia	2.6 (1.9)	13
Mysidopsis bahia	23.0 (7.2)	13
TM
MICROTOX	8.0 (6.4)	13
-170-

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ACUTE TOXICITY OF SECONDARY EFFLUENT
1000.0
1000.0
100.0
t—i—i—i—i—i—r~i—i—r
i—r- r r i i—i—i—p—i—i—i—i—i—i—r-t—i—i—i—i i i—i t— i—i "i
Apr MayJun Jul Aug Sep Oct Nov Jan
MICROTOX Mysidopsis Ceriodaphnia
ACUTE TOXICITY OF SECONDARY EFFLUENT
MEAN
LC50/EC50 (S.D.) N
Ceriodaphnia dubia 6.3 (4.6)
45
Mysidopsis bahia
47.6 (23.1)
44
MICROTOX
TM
79.3 (23.4)
40
-171-

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CHRONIC TOXICITY OF
PRIMARY AND SECONDARY EFFLUENT
NOEC MEAN ChV MEAN
(S.D.)	(S.D.) N
Ceriodaphnia dubia
Primary Effluent 0.8 (1.1) 1.2 (1.8) 12
Secondary Effluent 2.3 (1.6) 2.8 (2.1) 45
PERCENT TOXICITY REDUCTION BY
THE PATAPSCO WWTP
MEAN (S.D.) N
MICROTOX™	87.7 (12.2)	37
5-minute EC5q
Mysidopsis bahia 55.5 (16.8)	12
96-hour LC 59
Ceriodaphnia dubia
48-hour LC50	60.7(30.4)	13
7-day ChV	62.5 (31.1)	12
-172-

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SUMMARY OF TOXICITY RESULTS
® Ceriodaphnia dubia was the most
sensitive species
• Significant correlation of Ceriodaphnia
dubia and Mysidopsis bahia
o Percent toxicity reduction ranged
from 50-90%
PLANT PERFORMANCE EVALUATION
Primary Treatment Did Not Reduce
Influent Toxicity
Increases in Acute Effluent Toxicity
Occurred During Reduced Plant Performance
Performance and Operations Were Not a
Major Cause of Effluent Toxicity
-173-

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TIME LETHALITY TE8T
ON WHOLE 8AMPLE
FILTERED
(1.0um) 8AMPLE

I AERATED FOR |
1 HOUR
48-HOUR
ACUTE TEST
UNAERATED
48-HOUR
I ACUTE TE8TI
UNAERATED
48-HOUR
I ACUTE TE8TI
AERATED
I TIME LETHALITY
OR 48-HOUR
TEST
RAI8E pH>11
! AERATE FOR 1 HR.
AND NEUTRALIZE
TIME LETHALITY
TEST
COS) COLUMN I
EXTRACTION |
%
-174-

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TIE PHASE I RESULTS
SECONDARY EFFLUENT 10 DECEMBER 1986
Whole Aerate Filter NH3-N C-18 Cation Anion Residual
Treatments
TIE PHASE I RESULTS
SECONDARY EFFLUENT 23 JULY 1986
70
Bebo
30
O
10
48-hour LC50
Theoretical LC50
1
100
90
80
70
60
50
40
30
20
10
0
Whole Aerate Filter	C-18 Cation Anion Residual
Treatments
-175-

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TIE PHASE I RESULTS
PRIMARY EFFLUENT 23 JULY 1986
100
90
80
70
60
50
40
30
20
10
0
Whole Aerate Filter	C-18 Cation Anion Residual
Treatments
TIE PHASE I RESULTS
SECONDARY EFFLUENT 23 JULY 1986
3 70
"5 80
|2 50
•	40
|2 30
O 20
C 10
•
S o
25 50 75 80 85 90 95 100
Percent Methanol Fractions From C-18 Column
-176-

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TIE PHASE I RESULTS
.SECONDARY EFFLUENT 10 DECEMBER 1986
S. 25 50 75 SO $5 90 95 100
Percent Methanol Fractions From C-18 Column
REFRACTORY TOXICITY
ASSESSMENT (RTA)
•	Collect Sewer Samples
•	Batch Simulation of Activated Sludge
Treatment Process
•	Measure Batch Effluent Toxicity
•	Rank Sources by Relative Toxicity
Loading
-177-

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DESCRIPTION OF INDIRECT
INDUSTRIAL DISCHARGERS
INDUSTRY
CODE	INDUSTRY PRODUCTS
A Organic Chemicals and Pesticides
B Detergent Alkylates, Hydrotropes, and
Petroleum Intermediates
C Emulsifiers, Surfactants, and Specialty
Monomers
D Organic and Inorganic Chemicals
E Washdown of Chemical Transport Trucks
RTA RESULTS - INDUSTRY B
Percent


Ceriodaphnia
Industrial
MICROTOX
™ ^C 50
) Time Lethality (TU)
Wastewater
Influent
Effluent
Influent
Effluent
100
45.7
100
76.0
22.9
75
65.4
100
88.1
24.5
50
100
100
100
25.5
25
82.8
—
100
26.2
10
100
—
—
—
RAS
100

10.7

-j 78-

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RTA RESULTS - INDUSTRY D
Percent


Ceriodaphnia
Industrial
MICROTOX
™ 
-------
TIE PHASE I RESULTS
INDUSTRY A -12 DECEMBER 1986
o
o
ID
o
O
X
¦
100
90
80
70
60
50
40
30
20
10
0
48-hour LC50
Theoretical LC50
I
100
80
80
70
80
SO
30
20
10
0
Whole Aerate Filter NH3-N C-18 Cation Anion Residual
Treatments


-O 100
3
¦o
90
¦
80
o
70
o
IO
80
o
50
.1
40
3
30
©
20
i
CO
10

0
TIE PHASE I RESULTS
INDUSTRY A - 12 MARCH 1987
Whole Aerate Filter NH3-N C-18 Cation Anion Residual
Treatments
-180-

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TIE PHASE I RESULTS
INDUSTRY E - 26 MARCH 1987
Whole Aerate Filter NH3-N C-18 Cation Anion Re8idual
Treatments
TIE PHASE I RESULTS
INDUSTRY A - 12 MARCH 1987
2 o
® 25 SO 75 80 8S >0 96 100
Percent Methanol Fractions From C-18 Column
-181-

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TOXICITY IDENTIFICATION EVALUATION
OF RTA EFFLUENTS
INDUSTRY PRINCIPAL TOXIC FRACTION
A
NON-POLAR ORGANICS

RESIDUAL TOXICITY
C
NON-POLAR ORGANICS

RESIDUAL TOXICITY
D
NON-POLAR ORGANICS

RESIDUAL TOXICITY
E
ANIONS
CONCLUSIONS
The WWTP Achieved Significant
Toxicity Reduction; However,
Substantial Acute and Chronic
Toxicity Remained
Effluent Toxicity Was Not Caused
By Poor Treatment Operation Or
Performance
-182-

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CONCLUSIONS
(Continued)
Non-Polar Organic Compounds
Appear To Be The Principal Effluent
Toxicants
Acute Toxicity To Ceriodaphnia Was
Largely Associated With Particles
> 0.2 um
CONCLUSIONS
(Continued)
RTA Was An Effective Tool For
Identifying Contributors To The
WWTP's Effluent Toxicity
-183-

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RECOMMENDATIONS
Test Enhanced Solids Removal Techniques
(e.g. Coagulation/Precipitation or Filtration)
For Sorbable Toxicity Reduction
Additional RTA Testing to Identify Sources
Contributing to the WWTP Toxicity
Pass-Through
-184-

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TRE INDUSTRIAL PROTOCOL
TRE INDUSTRIAL
PROTOCOL -- AN OVERVIEW
Toxicity Reduction Evaluation
(TRE)
An investigation conducted within a plant
or municipal system to isolate the sources
of effluent toxicity and/or determine the
effectiveness of pollution control options
in reducing the effluent toxicity.
A TRE is a step-wise process
consisting of:
« Evaluation of existing site-specific information
® Toxicity characterization/identification evaluation
•	Confirmation
° Source Evaluation
•	Toxicity reduction method evaluation
•	Method selection and implementation
•	Follow-up monitoring
-185-

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TREs
Toxicants may never actually be identified -
perhaps only characterized
These are multidisciplinary investigations -
environmental toxicology, environmental
chemistry, environmental engineering,
process engineering, wastewater treatment
TREs
•	These investigations have proceeded
beyong the research state - the
objectives are achievable
•	The knowledge base is increasing
rapidly
•	The process can be simple or
complex, depending upon the
characteristics of the effluent
Industrial TREs
The industry employees are important team
members
Isolation and identification of toxic
contributors is more straightforward than in
municipal TREs
Indirect discharges may require a TRE
-186-

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TRE Flowchart
Evaluation of
Existing Site-Specific
Information
Information Acquisition
•	Plant and process descriptions
•	Influent and effluent physical/
chemical data
•	Effluent toxicity data
•	Instream biological data
-187-

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Housekeeping
•	Materials handling
•	Spill control
•	Facility cleanliness
•	Waste handling, storage
and disposal
•	Environmental awareness
Treatment System
•	Design characteristics
•	Operating efficiency
•	Influent characteristics
•	Effluent characteristics
Chemical Use
•	Process chemicals
•	Biocides
•	Cleaning operations
•	MSDS review
Products Manufactured
• Raw materials
® By-products
-188-

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Episodic Events
•	Intentional or unintentional releases of
toxics to the wastewater treatment
system
•	Accidental spills of toxics
Two TIE Approaches Are Possible
Toxicity Identification Evaluation
Objective: Identify cause(s) of
final effluent toxicity
Ideally: Identify specific chemical(s)
^ Not: Characterize the toxicants and
identify group(s) or fraction(s)
of chemicals
Specific Toxicant Identification
Identify toxicants so that they can
be eliminated or reduced at the source,
either by substitution or pretreatment.
-189-

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TIE STRATEGY
•	Evaluation of Variability
•	Performance of Toxicity
Fractionation / Characterization
•	Identification of Specific Toxicants
•	Confirmation of Identifications
Effluent Variability
•	Volume
•	Constituents
-190-

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Effluent Variability
Test Date. Days
Effluent Variability
Hours
-19J-

-------
Effluent Characterization
Baseline
Toxicity Tests
Toxic Effluent Sample
Degradation
Tests
Reducing
Agent Test
Chelation
Test
Air Stripping
Test
Filtration
Test
Cte Solid Phase
Extraction Test
Acid Base Neutral
Acid Base Neutral
Toxicity Treatment
Characterize whole effluent toxicity
and then develop treatment procedures
to reduce the toxicity to acceptable
levels.
Toxicity Reduction
Method Evaluation
•	Eliminate at the source
•	Treatment of final effluent
-192-

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Source Elimination
Chemical substitition
Process modification
Treatment of process streams
(pretreatment)
Eliminate the process
Treatment of Final Effluent
Identify possible treatment methods
Modify or add to present system
Design and construct new
treatment system
TOXICITY PERSISTENCE
« Can be useful in sourcing toxicants and
in identifying treatment options
Follow-Up Monitoring
•	Optimize toxicity reduction
method
•	Confirm toxicity reduction
•	Assess acceptability of
toxicity variability
-193-

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Factors to Consider
Before Initiating a TRE
DILUTION WATER OPTIONS
•	Receiving Water
•	Laboratory Water
•	Reconstituted Water
Proper Sample Collection,
Shipment and Storage Is Critical
to Maintain Realistic Effluent
Characteristics
-194-

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EFFLUENT COLLECTION
SHIPPING AND STORAGE
.	Grab Sample vs. Composite
•	Shipped on Ice in the Dark
•	Overnight Courier
•	Stored Until Use at About 4° C
Species Sensitivity
•	Temporal variability
•	Very important in establishing and
attaining the goals of the TI/RE
The Development of a Sufficient
Database Is the Only Way to Ensure
That Inappropriate Treatment
Costs Are Avoided
-195-

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On-Site Cooperation Is Essential
•	With an industry it is virtually impossible
to Identify the source of the problem(s)
without their cooperation.
•	Frequently an on-site person assigned
to the TI/RE full-time will be very
valuable.
Work Cooperatively With the
Regulatory Authority
•	Agree upon a reasonable goal.
•	Work together to accomplish the goal.
•	Assure that the goal does not change
in the process of conducting the TRE.
•	Flexibility in the schedule is the key.
•	Maintaining a good faith effort is important
•	H is always easier to be successful if a good
relationship is maintained between permittee,
contractor, and regulatory authority.
Increased Level of Environmental
Awareness/Consciousness
# A requirement If long-term and widespread success If to
be achieved.
-196-

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Summary
•	A defined TRE target Is essential
•	Generalized methodologies should
be developed and applied
•	Flexibility In design, implementation
and schedule
•	TREs should be facility-specific
•	Effluent variability Influences
design of TRE
•	Confirmation of causes of toxicity
important
•	Cooperation among all participants
critical
Chlorine Toxicity
•	Pass-through cooling water
toxicity
•	POTW disinfection toxicity
Unionized Ammonia Is Primarily
Responsible for Ammonia Toxicity,
and Both Temperature and pH
Control Percent Unionized Ammonia
-197-

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Ammonia Toxicity is Difficult to
Interpret in Shipped Effluent
Samples Collected in Cold Weather
Heating the Municipal Effluent From
(5 - 10° C) up to 25° C Increases
the Percent Unionized Ammonia
Substantially, as Does an
Increase in pH From 7.0 to 8.0
The Result Is a More Toxic Effluent
Adjusting Test Temperatures and pH for
Fish Bioassays Is Possible and
Can Provide for More Environmental
Realism in Toxicity Tests
Toxicity Resulting From Polar
Organics Presents Difficult
Problems in TREs
-198-

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Fractionation/Characterization Procedures
Do Not Necessarily Produce Absolute
Separation of Classes of Toxicants
Effluent Matrix Variability
Can Also Affect the
TRE Process
In TRE Work the Results of Chemical
Analyses Need to Be Carefully Interpreted
Sample Preparation Procedures
Deserve Special Scrutiny
-199-

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Section 7
Toxicity Identification Evaluation (TIE) Overview
-201-

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TOXICITY IDENTIFICATION
EVALUATION OVERVIEW
PRIORITY POLLUTANT ANALYSES
• Do not represent all toxic
chemicals
Two Approaches to Toxicity Reduction Evaluations:
Toxicity Investigation Evaluation
(Causative Agent Approach)
Effluent Toxicity Treatability
Both the Treatability & Identification
Approaches Utilize Physical-Chemical
Characteristics of Toxicants.
TIE BASED ON TWO PRINCIPLES
Concentrate
Separate
-203-

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ai «)4ns7<>niniR n
AhuwMnrf
TtMinnnn
snoimon
4nnnnon
JL-JL




I«vt» {mMt )
THREE MAJOR PARTS OP THP TIP
Characterization
Identification
Confirmation
Identify General Nature of Toxicant(s)
--	Solubility
-	Volatility
••	Decomposabillty
••	Complexabllity
••	Filterabillty
••	Sorbability
-204-

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I TOXICITY CHARACTERIZATION
MANIPULATION TYPE CHARACTERIZATION
pH adjustment
Filtration and pH
adjustment
Purging and pH
adjustment
C-18 SPE and pH
adjustment
Thiosulfate and
EDTA
Toxic form altered
Suspended solids
Solubility changes
Volatiies
Non-polar compounds
Oxidants and metals
Following Characterization,
the choice of
Treatability or Identification
is best made.
II. TOXICITY IDENTIFICATION
GROUP
Non-polar organics
Metals
Ammonia
Surfactants
Polar organics
Volatiies
-205-
APPROACH
SPE, HPLC, GC/MS
AA, Ion exchange,
chelation
pH manipulation,
zeolite
Bubble removal

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FINAL CONFIRMATION
1.	Toxicity vs. Concentration
Correlation
2.	Symptoms
3.	Spiking Effluent
4.	Toxicity Mass Balance
5.	Other Species
6.	Spiking Fractions
7.	Misc.
PH
Hardness
Tissue Uptake
-206-

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2	4
TOXIC UNITS OF SUSPECT TOXICANTS
Toxic Components of a POTW Effluent
w _
t 3
z
3
o
* 2
O *
Who** Effhwnt
WE Filtered
•Total PMUclds
I I V
I I I I I I
EFFLUENT SAMPLE
Presence of toxic concentrations
does not prove
the cause of toxicity.
-207-

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Amount of industrial
contributions not
related to toxicity.
A GOOD TIE PLAN CONSISTS OF
SEVERAL ASPECTS:
•	Use of broad "Characterization" steps.
•	Effluent variability conaidered
•	Toxicity tracked with analyaea.
•	Analytical capability is broad.
•	Reliance on the GC/MS Is realistic
•	Choice of test species logical.
•	Toxicity tests are streamlined.
•	Supply of test organiams is not limiting.
•	Team work approach is built In.
Choice of Test
Species for TIE?
-208-

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Does Objective of TIE
Include Resident Species?
TRIGGERS
FOR
TRE's
CHRONIC
TIE METHODS
LIMITED
EPA ACUTE
METHODS
NOT APPLICABLE
CHRONIC TRE
TREATABILITY METHODS
COMPLETE
-209-

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CHOOSING
A
CONTRACT FIRM
TOXICANT PATTERNS
ARE
EMERGING
-210-

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LEGEND
~	< .10 ug/l
~	0.10-0.25
¦ >0.25
— Not datemtoed
Figure 2. Frequency of dazinon occurrence in POTW
effluents from around the United States.

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64 Sites Evaluated
6 Lacked Acute Toxicity
58 TIE'S
Summary of NETAC TIE'S
Total Samples Evaluated	58
Successful Phase I	39 of 40
Successfully Determined
if Toxicant is X
(e.g., ammonia)?	18 of 18
Successful for Phase II
and/or III	16 of 18
Phase I Findings:
pH dependent	3
Inorganics	15
Oxidants	9
Non-polars	25
Volatiles	1
-212-

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Some Toxicants Identified
thru February 1989
Discharge Type
Industrial
Compound^
zinc, non-polar organics
¦	salinity (TDS)
- ammonia
¦	nickel
Municipal	zinc, non-polar organics
ammonia, diazinon, malathion
nickel
diazinon, chlorfenvinphos
diazinon, dichlorovos
-	ammonia, non-polar organics
diazinon, non-polar organics
-	diazinon
detergents
Other:
Ambient
Ambient
carbofuran, methyl parathion
diazinon
Elutriates
Elutriates
ammonia
manganese
One to three toxicants
have usually been
most of the problem.
-213-

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Toxicity often occurs
at
ug/L concentrations.
In a 35 mgd plant
1 ug/L s 0.3 lbs/day.
With 90% removal
1 ug/L = loading of about 3 lbs/day.
TOXICANTS AT ug/L CONCENTRATIONS:
•	Can't be found by examination
of loadings
•	Can't be related to conventional
pollutants
•	Are not likely to be related to
flow
CONCLUSIONS:
1.	TIE/TRE's are enjoying good success.
2.	Many contractors are able to do TIE'S.
3.	Phase I, characterization, is rapidly
becoming routine.
4.	Trends are emerging.
5.	Solutions are being found.
6.	Causes of toxicity are often few.
7.	The cause of toxicity is often not
industrial.
-214-

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NON POLAR ORGANIC
Toxicity Removed by C18
Toxicity Eluted from C18
AMMONIA
More toxic at high pH
Fathead minnows more sensitive
Total ammonia >10-20 mg/L
SURFACTANTS
Toxicity Reduced by:
Filtration
Aeration
C18
Toxicity degrades in plastic faster than in glass
-215-

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CATIONIC METALS
Removal/Reductions in Toxicity by:
EDTA
Ion Exchange
C18
Filtration
Thiosulfate
Toxicity pH sensitive
OXIDANTS
Toxicity reduced by:
Thiosulfate
Aeration
Aging
TRC measurable at effluent LC50
TDS
Conductivity > 3000 at LC50
no toxicity loss from:
Aeration
C18
Filtration at pHi
Thiosulfate
Not pH sensitive
Ion exchange reduces
Fathead minnows less sensitive
-216-

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Section 8
Guidelines and Review Criteria for TRE Plans
-217-

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GUIDELINES AND REVIEW
CRITERIA FOR
TRE PLANS
GUIDELINES
FOR
TOXICITY REDUCTION
EVALUATION PLANS
A DISCHARGER WILL BE REQUIRED TO SUBMIT A
TRE PLAN TO THE REGULATORY AUTHORITY
IN ORDER TO PROVIDE:
•	A DESCRIPTION OF THE STUDY PLAN
•	A SCHEDULE FOR CONDUCTING SPECIFIC TASKS
AND REPORTING THE RESULTS
•	RELEVANT BACKGROUND INFORMATION ON THE
FACILITY
•	WHO WILL BE CONDUCTING THE EVALUATION
THE TRE PLAN SHOULD CLEARLY ESTABLISH: |
•	SPECIFIC OBJECTIVES (TARGET) OF THE STUDY
•	MONITORING TEST CONFIRMATION OF REDUCTION
•	THE SCHEDULED COMPLETION DATE AND MILESTONES
¦219-

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DEVELOPMENT OF THE TRE PLAN IS SOLELY THE
RESPONSIBILITY OF THE DISCHARGER
• 1 TO 3 MONTHS WILL BE SUFFICIENT FOR PLAN
DEVELOPMENT
COMMUNICATION AND COOPERATION IN TRE PLAN
DEVELOPMENT AND REVIEW WILL HELP ENSURE A
AND GOOD FAITH EFFORT TO ACHIEVE
THE TRE OBJECTIVE
U.S EPA DOES NOT RECOMMEND -
TRE PLANS BE FORMALLY APPROVED
UNLESS REQUIRED BY STATE REGULATIONS
FLEXIBILITY IN DESIGNING AND CONDUCTING A TRE
WILL BE NECESSARY
•	SOME DECISIONS ON THE MOST APPROPRIATE
APPROACH MUST BE BASED ON THE RESULTS
OF THE INITIAL STEPS OR TIERS OF THE TRE
•	BE WARY OF TFlEa THAT MAY BECOME RESEARCH
PROJECTS INVESTIGATING NEW AND UNPROVEN
METHODS AND PROCEDURES
•	USE THE EPA GUIDANCE AS BASIS FOR REVIEW
•	REQUEST REASONS AND DOCUMENTATION FOR ANY
MODIFICATIONS OR ALTERNATIVE APPROACHES
THINGS TO AVOID
•	RELIANCE ON PRIORITY POLLUTANT SCANS
•	RESEARCH ON SURROGATE PARAMETERS TO
CORRELATE WITH AQUATIC ORGANISM TOXICITY
TESTS
•	LACK OF PERIODIC PROGRESS REPORTS
•	CONFIRMATION STEP NOT CARRIED OUT FOR
IDENTIFICATION OR TREATABILITY STUDIES
•	PROPOSED SCHEDULE FOR TRE IS ARRIVED AT BY
ADDING TOGETHER TIMES FOR TRE STEPS THAT ARE
USUALLY ALTERNATIVES OR ELSE CONDUCTED
CONCURRENTLY
•	INCLUSION OF STUDIES NOT DIRECTLY RELEVANT TO
CONDUCTING THE TRE AND ACHIEVING THE TRE
OBJECTIVE
-220-

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THINGS TO ENCOURAGE"!
© INTERIM REPORTS WITH SUMMARY DATA AND NEXT
STEPS TO BE CONDUCTED
e SPECIFICATION OF THE NUMBER, TIMING OF SAMPLES,
AND THE SAMPUNG POINTS
o THE SPECIFIC PROCEDURES TO BE FOLLOWED IN THE
VARIOUS COMPONENTS OF THE EVALUATION
® PRELIMINARY OR MINIMUM ESTIMATES OF HOW MANY
OF EACH ANALYSIS WILL BE CONDUCTED
o INDICATION THAT INVESTIGATORS HAVE
INTERDISCIPLINARY EXPERTISE
• THOROUGH QA/QC PRACTICES INCORPORATED IN ALL
CONFIRMATION STEPS
EVALUATION CRITERIA}
® ARE THE OBJECTIVES OR TARGETS OF THE TRE
CLEARLY AND ACCURATELY STATED?
•	ARE THE SCHEDULE AND MILESTONES FOR
ACCOMPLISHING THE TASKS DESCRIBED IN THE STUDY
PLAN?
•	ARE THE FINAL REPORT, PROGRESS REPORTS AND
MEETINGS WITH THE REGULATORY AUTHORITY
INCLUDED AS PART OF THE SCHEDULE?
•	ARE THE APPROACHES OR METHODS TO BE UTILIZED
DESCRIBED TO THE EXTENT THAT IS POSSIBLE PRIOR
TO REACHING DECISION POINTS AND WITHOUT THE
RESULTS AND DATA THAT WILL BE COLLECTED IN THE
INITIAL STEPS OR TIERS OF THE TRE?
o HAS THE AVAILABLE GUIDANCE BEEN UTILIZED IN THE
DESIGN OF THE TRE AND THE DEVELOPMENT OF THE
TRE PLAN?
© DOES THE TRE PLAN ALLOW A SUFFICIENT AMOUNT OF
TIME AND APPROPRIATE LEVEL OF EFFORT FOR EACH
OF THE COMPONENTS OF THE STUDY PLAN?
•	DOES THE TRE PLAN SPECIFY WHAT RESULTS AND
DATA ARE TO BE INCLUDED IN THE INTERIM AND FINAL
REPORTS?
® DOES THE TRE PLAN PROVIDE FOR ARRANGEMENTS
FOR ANY INSPECTIONS OR VISITS TO THE FACILITY OR
LABORATORY WHICH ARE DETERMINED BY THE
REGULATORY AUTHORITY TO BE NEEDED?
•	ARE THE TOXICITY TEST METHODS AND ENDPOINTS TO
BE UTUZED SPECIFIED OR REFERENCED?
-221-

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•	ARE OPTIMIZATION OF EXISTING PLANT/TREATMENT
OPERATIONS AND SPILL CONTROL PROGRAMS PART
OF THE INITIAL STEPS OF THE TRE?
•	DOES THE TRE PLAN INCLUDE A TIMELINE
(HORIZONTAL BAR GRAPH) WHICH CLEARLY
ILLUSTRATES THE TIMEFRAME FOR CONDUCTING THE
SPECIFIC COMPONENTS OF THE TRE (AS DESCRIBED
IN THE GUIDANCE) AND ANY OVERLAP OF THESE
COMPONENTS?
•	DO THE SUBSEQUENT TESTS AND EVALUATIONS BUILD
ON THE PREVIOUS RESULTS AND PROCEED BY
NARROWING DOWN THE POSSIBILITIES IN A LOGICAL
PROGRESSION?
•	ARE ALL TEST RESULTS ANALYZED AND USED TO
FOCUS ON THE MOST EFFECTIVE APPROACH FOR
SUBSEQUENT SOURCE INVESTIGATIONS, TREATABILITY
STUDIES, AND CONTROL METHOD EVALUATIONS?
^EVALUATION CRITERIA FOR lils]
•	ARE THERE BROAD CHARACTERIZATION STEPS?
•	IS EFFLUENT VARIABILITY CONSIDERED?
•	IS TOXICITY TRACKED WITH ANALYSES?
•	IS THE ANALYTICAL CAPABILITY BROAD?
•	IS RELIANCE ON GC/MS REALISTIC?
•	IS CHOICE OF TEST SPECIES LOGICAL?
•	IS TOXICITY TESTING STREAMLINED?
•	IS SUPPLY OF TEST ORGANISMS ADEQUATE?
•	IS TEAM APPROACH BUILT IN?
•	IS CONFIRMATION INCLUDED?
-	MASS BALANCE	- OTHER SPECIES INCLUDED
-	CORRELATION	- SPIKING
-SAMPLING OVER TIME - BIOAVAILABILITY
-222-

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Section 9
TRE Case Studies and Summary
-223-

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TOXICITY REDUCTION
EVALUATION ABSTRACTS
AND CASE STUDIES
-225-

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MARTINEZ MANUFACTURING COMPLEX
SHELL OIL COMPANY
MARTINEZ,CALIFORNIA
TOXICITY RESEARCH 1976-1985
REFINED PETROLEUM PRODUCTION FACILITY
PRODUCES GASOLINE. DIESEL FUEL. LUBE OILS
AND GREASE
uaidciiiiiui

\
SHELL
TREATMENT PROCESSES

• OIL/WATER SEPARATION

•BIOLOGICAL OXIDATION

• SECONDARY CLARIFICATION

9 TERTIARY FILTRATION

>-

EARLY STUDIES
INITIAL FRAGTIONATION/CHARACTERIZATION
SHOWED OIL AND GREASE AND AMMONIA
TO BE PARTIALLY RESPONSIBLE FOR
TOXICITY
PRIOR TO CONFIRMATION, ACUTE
TOXICITY DISAPPEARED
-226-

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t
mamt
SUBSEQUENT METHOD
• EFFLUENT OCCASIONALLY
ACUTELY TOXIC
• CORRELATE OBSERVED EFFLUENT
TOXICITY TO MANUFACTURING
PROCESSES
• RELATE CHANGES IN PROCESSES TO
PERIODS OF TOXICITY/ NO TOXICITY
• NARROW SCOPE OF WORK TO
FOCUS ON PROCESSES WHICH
CORRELATE WITH EFFLUENT
TOXICITY
r

SUSPECTED TOXICANTS:

AMMONIA

OIL AND GREASE

•NAPHTHENIC ACIDS

DIALLYLAMINE PROCESSES

¦SEVERAL AMINE COMPOUNDS

POLYETHYLENEIMINE

¦HOCCULATING AGENT

-227-

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AMINES
CAUSE OF TOXICITY?
-CONVERTED TO AMMONIA DURING
BIOTREATMENT?
'PASS-THROUGH AT HIGH
CONCENTRATIONS?
•INHIBIT NITRIFICATION OF AMMONIA?
CONTROL METHOD (1976-9):
¦BYPASS WATER SCRUBBER:
INCINERATE ETHYLENEDIAMINE
DIRECTLY
-INSTALL ION ELECTRODE MONITOR:
MONITOR AMINE/AMMONIA
CONCENTRATIONS. AT HIGH
CONCENTRATIONS. RERUN
TREATMENT
\
SHELL
AMMONIA
SUSTAIN NITRIFICATION
BMPs
•CONTROL OF SLUDGE AGE
¦CONTROL PH
•AVOID INHIBITORY ADDITIVES (SUCH AS PEI)
-GREATER EMPHASIS ON SPILL CONTROL
-228-

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POLYETHYLENEIMINE
(PEI)
CAUSE OF TOXICITY:
¦free pei toxic to fish
¦INHIBITS DEGRADATION OF
OIL AND GREASE
•INHIBITS NITRIFICATION (RESULTING
IN HIGH CONCENTRATIONS OF
NAPHTHENIC ACIDS AND AMMONIA)
CONTROL METHOD:
¦REPLACE PEI WITH A DIFFERENT.
LESS TOXIC FLOCCULATING
AGENT
OIL AND GREASE
(NAPHTHENIC ACIDS)
SOURCE:
-CRUDE OIL DESALTER (NAPHTHENIC
ACIDS PARTITION INTO WATER PHASE)
CONTROL METHOD:
¦BRINE DEOILING UNIT ADDED TO
REDUCE CONCENTRATION OF
NAPHTHENIC ACIDS PARTITIONING
INTO WATER PHASE
•PACT ADDITION TO ACTIVATED
SLUDGE FOLLOWING SPILLS OR
UPSETS
-229-

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f
01
\
XX HAVEN
GLEN RAVEN MILLS
ALTAMAHAW, NORTH CAROLINA
FEBRUARY 1985- MARCH 1986
REQUIRED TO MEET 48 HOUR ACUTE
STATIC LC50 OF >90% ON
DAPHNIA PULIX


<



f ' N
OLE* IAW


PROCESS:


DYES PANTYHOSE WITH ACID
AND DISPERSE DYES


CHEMICALS USED:


DYESTUFFS


SURFACTANTS


CHELATING AGENTS


FABRIC SOFTENERS




CUR lUVCJt
TREATMENT PROCESSES:
EQUILIZATION
ACTIVATED SLUDGE TREATMENT
CLARIFICATION
CHLORINATION
-230-

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GLCff RAVEN
TIER 1: CHEMICAL
COMPOUND
OPTIMIZATION
ELIMINATE/MINIMIZE CHEMICALS
WITH KNOWN TOXICITY AND
MINIMAL BIODEGRADABILITY
AlKYl PHENYL ETHOXYLATES 
-------
/	A
CLOT KAVTW
TIER 3: EFFLUENT
CHARACTERIZATION
TOXIC CONCENTRATIONS FOUND:
COPPER
NICKEL
ZINC
NONBIODEGRADED NONIONIC
SURFACTANTS
LINEAR ALCOHOL ETHOXYLATES
	\
OLtJf MAVIS
METAL REDUCTION
EXPERIMENT:
CATIONIC EXCHANGE RESIN
SUBSTANTIAL REDUCTION OF COPPER
AND ZINC
MINIMAL REDUCTION OF IRON
SOME REDUCTION OF CADMIUM.
CHROMIUM, LEAD AND NICKEL
EFFLUENT LC50
71.9% UNTREATED
•0.7% TREATED
METALS NOT LIKELY SOURCE OF
TOXICITY
HIGH CONCENTRATIONS PROBABLY
CHELATED
-232-

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EXTENDED BIOLOGICAL
TREATMENT EXPERIMENT
TREATMENT
FULLY DEGRADE AND TREAT
SURFACTANTS
ACTIVATED SLUDGE RENEWAL
(20% EVERY FIVE DAYS)
EXTENDED TREATMENT
EFFLUENT LC50
71.9% UNTREATED
>90% TREATED
f	N
aim iuvxn
CONCLUSIONS:
EFFLUENT IS NONTOXIC IF ADEQUATE
BIOLOGICAL TREATMENT IS RECEIVED
ADDITIONAL BIOLOGICAL TREATMENT
BIODEGRADES SURFACTANTS
AND ORGANICS
REDUCES COD LOADING
MAXIMUM FLOW OF WASTEWATER
SHOULD BE NO GREATER THAN 20% OF
TREATMENT FACILITY CAPACITY OR
EFFLUENT SHOULD RECEIVE 20% MORE
SLUDGE CONTACT TIME
CONCENTRATIONS OF TOTAL
RECOVERABLE METALS EXCEED ACUTELY
TOXIC CONCENTRATIONS BUT DO NOT
APPEAR TO BE CONTRIBUTING
SIGNIFICANTLY TO TOXICITY
(DUE TO CHELATION?)
-233-

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TRE SUMMARY
INFORMATION
SYGBTES UIBTR THE PSOS88BMS
898
WHERE IBEs QRBE SEEN ©ONE
-234-

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THE STUDY LOOKED AT 34 TREs - ALL CONDUCTED PRIOR
TO PUBLICATION OF EPA PROTOCOLS
TYPES OF FACILITIES REPRESENTED:
4 OIL REFINERIES
4 METAL INDUSTRIES
4	TEXTILES FACILITIES
8	CHEMICAL INDUSTRIES
9	POTWs
5	MISCELLANEOUS
• TREs RANGED FROM 3 MONTHS TO 2 YEARS DEPENDING
ON THE COMPLEXITY OF THE WASTESTREAM AND
VARIABILITY OF TOXICITY
TOXICANTS
TOXICANTS WERE IDENTIFIED AT 23 FACILITIES
¦PESTICIDES AND HERBICIDES AT 7 FACILIVES
-OTHER ORGANICS AT 12 FACILITIES
¦AMMONIA AT 6 FACILITIES
¦METALS AT 5 FACILITIES
V.
tea
-2313-

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TREATMENT
•
13 FACILITIES HAVE SELECTED AND
IMPLEMENTED TREATMENT
•
6 FACILITIES HAVE SELECTED BUT NOT
IMPLEMENTED TREATMENT
•
13 FACILITIES HAVE NOT SELECTED TREATMENT
©
2 FACILITIES HAVE CLOSED THE OUTFALL

- 1 DIVERTED TO A POTW
¦1 STOPPED PROCESS CAUSING TOXICITY
TREATMENT EFFECTIVENESS:
•OF THE 13 FACILITIES WHICH IMPLEMENTED
TREATMENT:
-6 HAVE MET PERMIT LIMITS
•2 HAVE REMOVED ACUTE TOXICITY BUT SVLL HAVE
CHRONIC TOXICITY
•r IS NOT MEETING PERMIT LIMITS
¦2 ARE UNKNOWN (TREATMENT RECENTLY INSTALLED)
• PRODUCT SUBSTITUTION, PACT AND SYSTEM
UPGRADES MOST COMMON TREATMENTS
SELECTED
-236-
« U.S. GOVERM-ENT PRINTING OFFICE' 1990 748-010/25007

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