A WET Tale: Toxicity of Complex

Effluents

Society of Environmental Toxicology and Society of Environmental Toxicology and ChemistryChemistry

WET Expert Advisory Panels

• Cooperative Agreement between U.S. USEPA Cooperative Agreement between U.S. USEPA and SETAC Foundation initiated in 1996.and SETAC Foundation initiated in 1996.

• Provides support on whole effluent toxicity Provides support on whole effluent toxicity (WET) issues through expert advisory panels (WET) issues through expert advisory panels (EAP).(EAP).

• WET EAP Steering Committee oversees and WET EAP Steering Committee oversees and guides actions of panels to address specific guides actions of panels to address specific issues (e.g., WET training and data issues (e.g., WET training and data interpretation). interpretation).

WET INTRODUCTION TOPICS COVERED IN

COURSE• Statutory and Regulatory Background of the WET Statutory and Regulatory Background of the WET

programprogram• Standards and Criteria in WET DevelopmentStandards and Criteria in WET Development• WET Methods, QA/QC, and Statistical ProceduresWET Methods, QA/QC, and Statistical Procedures• NPDES USEPA and State WET Permit DevelopmentNPDES USEPA and State WET Permit Development• Permittees Perspective on the WET ProgramPermittees Perspective on the WET Program• Compliance and Enforcement WET ProceduresCompliance and Enforcement WET Procedures• TRE/TIE Overview with Case HistoryTRE/TIE Overview with Case History

PURPOSE OF WORKSHOP

• Foster better understanding of scientific Foster better understanding of scientific underpinnings of WET as a toolunderpinnings of WET as a tool

• Provide overview of key implementation Provide overview of key implementation considerationsconsiderations

• Discuss current WET issues and Discuss current WET issues and concernsconcerns

STATUTORY OF WATER POLLUTION CONTROL

• Rivers and Harbors Act of 1899Rivers and Harbors Act of 1899

• Public Health Services Act of Public Health Services Act of 19121912

• Oil Pollution Act of 1924Oil Pollution Act of 1924

• FWPCA of 1948, 1956, 1961, FWPCA of 1948, 1956, 1961, 19651965

• FWPCA of 1972, 1977, 1987FWPCA of 1972, 1977, 1987

EARLY USE OF TOXICITY TESTING IN THE USEPA WET

PROGRAM• 1974, Region 4 conducted on-site acute tests1974, Region 4 conducted on-site acute tests

• 1976, selected industrial facilities conducted acute tests1976, selected industrial facilities conducted acute tests

• 1982, SETAC workshop evaluated USEPA WET testing1982, SETAC workshop evaluated USEPA WET testing

• 1983, began use of short-term chronic testing1983, began use of short-term chronic testing

• 1983, WET as a predictor of receiving water toxicity 1983, WET as a predictor of receiving water toxicity evaluated evaluated

• 1984, USEPA policy recommended use of WET limits in 1984, USEPA policy recommended use of WET limits in NPDES permits NPDES permits

• 1985, USEPA TSD for toxics control1985, USEPA TSD for toxics control

• 1991, USEPA revised the TSD for toxics control1991, USEPA revised the TSD for toxics control

ONGOING WET ACTIVITIES• SETAC Pellston WETSETAC Pellston WET

– unresolved scientific issuesunresolved scientific issues– needed researchneeded research

• National meeting to discuss WET National meeting to discuss WET implementation issuesimplementation issues – standardsstandards– permittingpermitting

• SETAC WET Expert Advisory PanelsSETAC WET Expert Advisory Panels

WET TESTING FOR AQUATIC LIFE PROTECTION

• USEPA promulgated test methods and guideline USEPA promulgated test methods and guideline test methodstest methods

• Acute and short-term chronic testsAcute and short-term chronic tests• Surrogate fresh and marine test species using Surrogate fresh and marine test species using

tropic level approach for sensitivity evaluationtropic level approach for sensitivity evaluation– fishfish

– invertebratesinvertebrates

– plantsplants

• Measures aggregate toxic effect of effluents or Measures aggregate toxic effect of effluents or receiving watersreceiving waters

WHY WET?• Implements National Policy “discharge of toxic Implements National Policy “discharge of toxic

pollutants in toxic amounts be prohibited”pollutants in toxic amounts be prohibited”• Permittee’s in compliance with chemical limits - Permittee’s in compliance with chemical limits -

effluent still toxic effluent still toxic • Directly implements States’ numeric or narrative Directly implements States’ numeric or narrative

criteria “no toxics in toxic amounts”criteria “no toxics in toxic amounts”• Whole effluent approach to toxics control for Whole effluent approach to toxics control for

protection of aquatic life uses acute and chronic protection of aquatic life uses acute and chronic toxicity tests to measure toxicity of effluentstoxicity tests to measure toxicity of effluents

• Cost effective approach in controlling toxic dischargesCost effective approach in controlling toxic discharges

INTEGRATED APPROACH TO WATER QUALITY-BASED

TOXICS CONTROL

• Chemical-specific approachChemical-specific approach

• Whole effluent toxicity Whole effluent toxicity approachapproach

• Biological criteria approachBiological criteria approach

INDEPENDENT APPLICABILITY POLICY

• Exceedance of any one approach Exceedance of any one approach is a violationis a violation

• Weight-of-evidenceWeight-of-evidence

WATER QUALITY STANDARDS AND WET

WATER QUALITY STANDARDS

Section 303 of the CWASection 303 of the CWA

WATER QUALITY STANDARDS THREE PARTS

• Designated usesDesignated uses

• Criteria (narrative and numeric) to Criteria (narrative and numeric) to protect designated usesprotect designated uses

• Antidegradation policyAntidegradation policy

DESIGNATED USES 40 CFR 131.10

ExamplesExamples– Recreation and aestheticsRecreation and aesthetics– Public water suppliesPublic water supplies– Propagation of fish and wildlifePropagation of fish and wildlife– Agricultural usesAgricultural uses– Industrial usesIndustrial uses– NavigationNavigation– Outstanding national resource waters (ONRW) Outstanding national resource waters (ONRW)

CRITERIA 40 CFR 131.11

• Numeric - individual pollutant or Numeric - individual pollutant or pollutant parameterspollutant parameters– copper, zinc, etc.copper, zinc, etc.

• Narrative - “free from…”Narrative - “free from…”– settleable solidssettleable solids– toxics in toxic amountstoxics in toxic amounts– aestheticsaesthetics– floatablesfloatables

ANTIDEGRADATION POLICY

40 CFR 131.12• Existing receiving water usesExisting receiving water uses

• Exceeds water qualityExceeds water quality

• Outstanding national resource water Outstanding national resource water (ONRW)(ONRW)

• Entrainment and thermal dischargesEntrainment and thermal discharges

IMPLEMENTATION POLICY

Explains how states implement the Explains how states implement the narrative criterion to establish effluent narrative criterion to establish effluent limitations to control WETlimitations to control WET

WATER QUALITY CRITERIA COMPONENTS

• Magnitude: maximum allowable Magnitude: maximum allowable concentration of WETconcentration of WET

• Duration: period of time over which the Duration: period of time over which the receiving water concentration is averagedreceiving water concentration is averaged

• Return frequency: designation of how often Return frequency: designation of how often the criterion may be exceeded without the criterion may be exceeded without impacting the organisms in the water bodyimpacting the organisms in the water body

USEPA HAS NOT DEVELOPED A 304(a) CRITERION FOR WET

USEPA RECOMMENDATIONS IN THE

TSD FOR ACUTE TESTS

• MagnitudeMagnitude == 0.3 TUa0.3 TUa

• DurationDuration == one hourone hour

• FrequencyFrequency == once in three once in three yearsyears

USEPA HAS NOT DEVELOPED A 304(a) CRITERION FOR WET

USEPA RECOMMENDATIONS IN THE TSD FOR CHRONIC

• MagnitudeMagnitude == 1.0 TUc1.0 TUc

• DurationDuration == 4 days4 days

• FrequencyFrequency == once in three once in three yearsyears

CRITICAL LOW FLOWS AND MIXING ZONES

• States WQS protect designated uses States WQS protect designated uses during critical low flow periodsduring critical low flow periods

• Mixing zones are allowable in some Mixing zones are allowable in some WQSWQS

STATUTATORY REQUIREMENTS FOR DEVELOPMENT OF WQBELs

FOR WET• CWA 101(a): objective - restore and maintainCWA 101(a): objective - restore and maintain• CWA 101(a)(1): national goal - eliminate CWA 101(a)(1): national goal - eliminate

dischargesdischarges• CWA 101(a)(3): toxic pollutants in toxic amountsCWA 101(a)(3): toxic pollutants in toxic amounts• CWA 301(a): requires permit to discharge CWA 301(a): requires permit to discharge

pollutantspollutants• CWA 301(b)(1)(C): permits must assure WQS CWA 301(b)(1)(C): permits must assure WQS

are metare met

REGULATORY REQUIREMENTS FOR DEVELOPMENT OF WQBELs

FOR WET

• 40 CFR 122.44(d)(1) 40 CFR 122.44(d)(1) requirements for developing WET requirements for developing WET limits limits

• Effluent characterization to Effluent characterization to determine reasonable potential determine reasonable potential (RP)(RP)

40 CFR 122.44(D)(1)BASIS FOR REASONABLE

POTENTIAL DETERMINATIONS AND DEVELOPMENT OF WQBELs

• List of factors for RP determinationList of factors for RP determination

• Limits developed for positive RPLimits developed for positive RP

• Positive RP for narrative criterion, Positive RP for narrative criterion, use of WET of chemical specific use of WET of chemical specific parameter(s) controlparameter(s) control

WET CONTROL POLICY JULY 1994

• Promote uniform, nationwide Promote uniform, nationwide compliance with statutory and compliance with statutory and regulatory requirements for the control regulatory requirements for the control of WETof WET

• Assist permit writers in implementation Assist permit writers in implementation these requirementsthese requirements

SUMMARY OF EIGHT STATEMENTS OF POLICY

(1) Basis for WET control(1) Basis for WET control

(2) Evaluation of discharges for RP(2) Evaluation of discharges for RP

(3) Evaluating RP(3) Evaluating RP

(4) Consequences of establishing RP(4) Consequences of establishing RP

- cont.- cont.

SUMMARY OF EIGHT STATEMENTS OF POLICY

(cont.)(5) WET monitoring(5) WET monitoring

(6) Compliance schedules in NPDES (6) Compliance schedules in NPDES permitspermits

(7) WET controls ammonia and chlorine(7) WET controls ammonia and chlorine

(8) WET controls POTWs(8) WET controls POTWs

CONCLUSIONS

• WET program highly successful in WET program highly successful in controlling toxic dischargescontrolling toxic discharges

• Useful tool in water quality control Useful tool in water quality control programsprograms

• Statutory and regulatory basis for WET Statutory and regulatory basis for WET is well establishedis well established

• The context WET of NPDES permitting is The context WET of NPDES permitting is defensibledefensible

Toxicity Test Methods

Acute and Short-term Chronic Toxicity Test

Methods for Effluents and Receiving Waters

MODULE OBJECTIVES

• Toxicity Testing Historical PerspectiveToxicity Testing Historical Perspective• Toxicity Test Methods for Effluents, Ambient Toxicity Test Methods for Effluents, Ambient

Waters, and Aqueous MediaWaters, and Aqueous Media• Key Parameters for TestingKey Parameters for Testing• Test Acceptability Criteria - what they meanTest Acceptability Criteria - what they mean• Test PrecisionTest Precision• Special IssuesSpecial Issues

HISTORY OF TOXICITY TESTING

• 1920’s: 1920’s: – Looked at test endpoints, such as immobilizationLooked at test endpoints, such as immobilization

• 1930/40’s: 1930/40’s: – Advantages of aquatic species for testing noted Advantages of aquatic species for testing noted

(size, short life span)(size, short life span)– Standard Methods included sewage & effluentsStandard Methods included sewage & effluents– Hart, Doudoroff and Greenbank published the Hart, Doudoroff and Greenbank published the

first aquatic toxicity methods.first aquatic toxicity methods.

1950s• Possible effects of chemicals and wastes on Possible effects of chemicals and wastes on

aquatic organisms.aquatic organisms.• Biologists observed striking differences in Biologists observed striking differences in

groups of organisms living in streams that groups of organisms living in streams that received waste materials and those that did not. received waste materials and those that did not.

• Enforcement cases, comprehensive basin plans, Enforcement cases, comprehensive basin plans, water quality criteria (WQC) developed by state water quality criteria (WQC) developed by state pollution control boards, Standard Methods pollution control boards, Standard Methods included “included “Examination of Industrial WastesExamination of Industrial Wastes””

1960s• FWPCA Water Quality Criteria established and FWPCA Water Quality Criteria established and

states required to adopt standards. states required to adopt standards. • Publication of paper on Measurement of Publication of paper on Measurement of

Pollutant Toxicity to Fish (Sprague).Pollutant Toxicity to Fish (Sprague).• Acute toxicity tests have been work-horse for Acute toxicity tests have been work-horse for

determining toxic effects since the late 60’s.determining toxic effects since the late 60’s.• FWPCA training manual FWPCA training manual Bioassay in Pollution Bioassay in Pollution

Analysis and ControlAnalysis and Control

1970s• USEPA was formed in December ‘70. USEPA was formed in December ‘70.

• FWPCA Amendments of ‘72 (Clean FWPCA Amendments of ‘72 (Clean Water Act) set forth a national pollution Water Act) set forth a national pollution program based on a discharge permit program based on a discharge permit system. system.

• Clean Water Act of 1977 revised; Clean Water Act of 1977 revised; included technology based approach to included technology based approach to effluent and criteria development. effluent and criteria development.

» continued

1970s - continued• NRDC/USEPA Consent Decree for 126 NRDC/USEPA Consent Decree for 126

priority pollutants. priority pollutants.

• Saw shift from limits of only BOD, DO, pH, Saw shift from limits of only BOD, DO, pH, suspended solids, temperature to toxics. suspended solids, temperature to toxics.

• ASTM committees develop aquatic ASTM committees develop aquatic toxicity test standardstoxicity test standards

• First USEPA effluent testing manual in First USEPA effluent testing manual in January 1978.January 1978.

1980s• Policy directed at “toxics” (vs BPT, BAT)Policy directed at “toxics” (vs BPT, BAT)• Development of short-term chronic testsDevelopment of short-term chronic tests

– Acute manual: 3rd editionAcute manual: 3rd edition– Freshwater chronic: 1st edition and 2nd editionsFreshwater chronic: 1st edition and 2nd editions– Marine and Estuarine: 1st editionMarine and Estuarine: 1st edition

• Field Validation of short-term chronic tests Field Validation of short-term chronic tests – Methods were being developed and field validatedMethods were being developed and field validated

– USEPA performed 8 site studies of toxicityUSEPA performed 8 site studies of toxicity tests and field tests and field assessments from 82-85.assessments from 82-85.

• TRE/TIE testing procedure developmentTRE/TIE testing procedure development

1990s• Notice of rulemaking for test manuals and Notice of rulemaking for test manuals and

public comment public comment • Toxicity Testing manuals revised:Toxicity Testing manuals revised:

* Acute: 4th edition* Freshwater chronic: 3rd edition* Marine/Estuarine: 2nd edition

* Testing guidance for West Coast Species

• TIE procedures revised and include TIE procedures revised and include chronic procedureschronic procedures

USEPA PROMULGATED BIOLOGICALMETHODS IN 40 CFR Part 136

• Methods must be followed as they are Methods must be followed as they are written!written!

• New permits and permit reissuance New permits and permit reissuance incorporate the protocols into the permit.incorporate the protocols into the permit.

• Existing permits do not get re-opened.Existing permits do not get re-opened.• Only the marine and estuarine chronic methods

are stayed for Pacific Coast dischargers.

TOXICITY TESTING APPROACH• Toxicity tests are designed to determine Toxicity tests are designed to determine

whether toxic chemicals are present in whether toxic chemicals are present in toxic amounts.toxic amounts.

• Not designed to be quantitative Not designed to be quantitative predictors of ecosystem responses--predictors of ecosystem responses--though many studies have though many studies have demonstrated significant associations demonstrated significant associations between toxicity test results and between toxicity test results and ecosystem impacts. ecosystem impacts.

SPECIES SELECTION FOR TOXICITY TESTING

• Sensitive species

• Ease of culturing and maintenance

• Readily available year-round

• Ecologically, commercially, and/or recreationally important

• Consistent and reproducible response

NO ONE SPECIES IS ALWAYS THE MOST SENSITIVE

Rank of SensitivitySpecies DDT Copper

Fathead minnow 34 20

Coho salmon 28 7

Chinook salmon 27 4

Rainbow trout 22 5

Bluegill sunfish 18 36

Largemouth bass 3 37

Daphnia magna 9 19

ACUTE TEST SPECIESFreshwater Species

InvertebratesCladoceran Ceriodaphnia dubia

Daphnia magnaDaphnia pulex

24, 48optional: 96

FishFathead minnowRainbow troutBrook trout

Pimephales promelasOncorhynchus mykissSalvelinus fontinalis

24, 48, 96 h

Marine/Estuarine Species

InvertebrateMysid shrimp Mysidopsis bahia 24, 48, 96 h

FishSheepshead minnowSilverside

Cyprinodon variegatusMenidia beryllinaMenidia menidiaMenidia peninsulae

24, 48, 96 h

SHORT-TERM CHRONIC TOXICITY METHODS: FRESHWATER

SPECIESTEST

DURATION ENDPOINT(S)

Cladoceran, Ceriodaphnia dubia

6-8 d Survival, Reproduction60% have 3 broods

Fathead minnow, Pimephales promelas

7-d

7-9 d

Survival, Weight

Survival, Percent hatch,Percent Abnormal

Green Alga, Selenastrum capricornutum

96 h Growth

SHORT-TERM CHRONIC TOXICITY METHODS: MARINE/ESTUARINE

SPECIESTEST

DURATION TEST ENDPOINT(S)

Mysid shrimp, Mysidopsisbahia

7-d Survival, Growth, Fecundity

Sheepshead minnowCyprinodon variegatus

7-d

7-9 d

Survival, Weight

Survival, Percent Hatch, PercentAbnormal

Inland Silverside, Menidiaberyllina

7-d Survival, Weight

Sea Urchin, Arbacia punctulata 1.5 h Fertilization

Red Macroalga, Champiaparvula

7-9 Cystocarp Production (fertilization)

METHOD MANUALS COVER

• Health and safetyHealth and safety• Quality assuranceQuality assurance• Facilities, equipment, suppliesFacilities, equipment, supplies• Test organismsTest organisms• Dilution waterDilution water• Effluent sampling and handlingEffluent sampling and handling• Test methodsTest methods• Report preparationReport preparation

METHOD MANUALS TERMINOLOGY

• Use of “Use of “maymay” or “” or “shouldshould””

• Use of “Use of “shallshall” or “” or “mustmust””

SELECTION OF TEST TYPE• Acute testsAcute tests

* static non-renewalstatic non-renewal* static renewalstatic renewal* flow-throughflow-through

• Short-term chronic methodsShort-term chronic methods* static non-renewalstatic non-renewal* static renewalstatic renewal* flow-throughflow-through

ACUTE TESTS• Duration - 96 h or lessDuration - 96 h or less• Endpoint - Endpoint - lethality, effect or no observed lethality, effect or no observed

adverse effect leveladverse effect level– lethality (LC)lethality (LC)– effect (EC)effect (EC)– NOAECNOAEC– Pass or FailPass or Fail

• Test Acceptability Criteria (TAC)Test Acceptability Criteria (TAC)– Typically survival in control, mean wt, Typically survival in control, mean wt,

mean youngmean young

SHORT-TERM CHRONIC TESTS

• Duration - 9 d or lessDuration - 9 d or less

• Endpoints - reproduction, fertilization, Endpoints - reproduction, fertilization, growth and sometimes lethalitygrowth and sometimes lethality

– ICp ICp

– NOECNOEC

– Pass or FailPass or Fail

SUMMARY OF TEST CONDITIONS

Each method has recommended test conditions:Each method has recommended test conditions:• test type and duration• temperature, light, DO, salinity• test chamber size and volume• species selection, age, feeding• dilution water• dilution series• sampling and holding requirements• test acceptability criteria (TAC)

SELECTION OF DILUTION WATER

• Dilution water may be either standard Dilution water may be either standard laboratory water or receiving water.laboratory water or receiving water.

• Choice of water is dependent on the Choice of water is dependent on the objectives of the test.objectives of the test.

BRINE AND BRINE CONTROLS

• Salinity can be adjusted with dry sea salts or Salinity can be adjusted with dry sea salts or hypersaline brine.hypersaline brine.

• Whenever salinity adjustment is necessary, Whenever salinity adjustment is necessary, brine controls must be used.brine controls must be used.

• Brine controls must meet TACBrine controls must meet TAC• Brine controls should be used in statistical Brine controls should be used in statistical

comparisons.comparisons.

EFFLUENT SAMPLING AND HOLDING• Sample types: Sample types:

• Grab vs. Composite• Grab samples cooled immediately Grab samples cooled immediately

• Composites cooled during collectionComposites cooled during collection

• If chlorinated, measure TRC of effluent at time of If chlorinated, measure TRC of effluent at time of sampling for grab and of composite when sampling for grab and of composite when sampling complete sampling complete

• Sample cooled to 4ºC and maintained during Sample cooled to 4ºC and maintained during shippingshipping

• Age of sample at test initiation or renewal Age of sample at test initiation or renewal <<36 h 36 h or not more than 72 hor not more than 72 h

TEST MEASUREMENTS NEEDED

• Measures to monitorMeasures to monitor»Dissolved oxygen cannot fall below 4 mg/l

(initialand final)»pH (initial and final)»conductivity»total residual chlorine »total hardness and alkalinity»salinity »temperature (arrival, initial, final, continuous))

TEST DILUTION SERIES

• USEPA recommends a USEPA recommends a multi-multi-concentrationconcentration test with a test with a minimumminimum of of 5 concentrations and control5 concentrations and control

TEST ACCEPTABILITY CRITERIA

• Valid vs. invalid testValid vs. invalid test

• Control criteriaControl criteria• minimum survival• minimum growth in weight or length,

reproduction, fertilization, etc.

COMPARISON OF TEST CONDITIONS: FRESHWATER ACUTE TESTS

Daphniamagna or

pulex

Ceriodaphniadubia

FatheadMinnow Trout

Temp (°C) 20 or 25 20 or 25 20 or 25 12

Age (d) < 1 < 1 1-14 15-30 RBT30-60 BT

Chamber 30 ml 30 ml 250 ml 5 L

Reps/Conc 4 4 2 2

No. Org/Conc. 20 20 20 20

Control TAC 90% 90% 90% 90%

COMPARISON OF TEST CONDITIONS: MARINE ACUTE

MysidSheepshead

MinnowInland

Silverside

Temp (°C) 20 or 25 20 or 25 20 or 25

Salinity (‰) 5 – 30 5 - 32 1 –32 (MB)

Age (d) 1 – 5 1 – 14 9 – 14

Chamber 30 ml 250 ml 250 ml

Reps/Conc 2 2 2

No. Org/Conc. 20 20 20

Control TAC 90% 90% 90%

COMPARISON OF TEST CONDITIONS: WEST COAST

MARINE ACUTE TESTS

Topsmelt MysidTemp (°C) 21 12 or 25Salinity (‰) 5 – 34 32 - 34Age (d) 10 – 30 3 – 5Chamber 250 ml 250 mlReps/Conc 2 2No. Org/Conc. 20 20Control TAC 90% 90%

CHRONIC FRESHWATER TEST CONDITIONS

C. dubia3 brood

Fathead7-d

Algae

Temp (°C) 25 25 25Age (h) <24 (8) <24 --Chamber (ml) 30 250 125 or 250

Reps/conc 10 3 - 4 3 - 4No. per conc 10 30 - 60 10,000 cells/ml/rep

Control TAC 80%3 brood in 60%

of ’s 15 yg/ surv

80% 0.25 mg

1 mil/cells w/EDTA2 mil/cells w/o

EDTA

NOTE: yg = young

CHRONIC MARINE TEST CONDITIONS

SheepsheadGrowth & Surv

SilversideGrowth & Surv

MysidSurv, Growth

Fecundity

Temp °C) 25 1 25 1 26 1Salinity 20-32 5-32 20-30Age < 24 h 7-11 d 7 dChamber (ml) 300 150 400Reps/conc 3-4 3 –4 8No. per conc 30-60 30- 60 40TAC control >80% >80% >80%

>0.60 mg >0.5 mg >0.2 mgfecundity if >50% of

controls developeggs

CHRONIC MARINE TEST CONDITIONS (continued)

ChampiaReproduction

ArbaciaFertilization

Temp (°C) 23 1 20 1Salinity 30 2 30 2Age -- --Chamber (ml) 200 20Reps/conc 3-4 3-4No. per conc 5 female tips and

1 males2000 eggs5 mil sperm

Control TAC >>80% survaverage of 10cystocarps per

plant

>>70-90% eggfertilization

WEST COAST SPECIES TEST CONDITIONS: CHRONIC TESTS

TopsmeltGrowth & Surv

MysidGrowth & Surv

Oyster, MusselEmbryo Larval

Temp (°C) 20 ± 1 13-15 ± 1 15 – 20 1Salinity 34 ± 2 34 ± 2 30 ± 1Age (d) 9 – 15 3 - 4 EmbryosChamber (ml) 600 150 30Reps/Conc 5 5 4No. per Conc. 25 25 150 – 300Control TAC >80% >75% >50 – 70% surv

>0.85 mg >0.40 mg >0.90 shell devel.

WEST COAST SPECIES TEST CONDITIONS: CHRONIC TESTS

AbaloneLarvalDevel.

UrchinEmbryo &

LarvalGiant Kelp

GerminationUrchinFert.

Temp (°C) 15 ± 1 15 ± 1 15 ± 1 12 ± 1Salinity 34 ± 2 34 ± 2 34 ± 2 34 ± 2

Age larvae Embryos Zoospore EmbryosChamber (ml) 600 30 600 16X 100 dish

Reps/conc 5 4 5 4No. per conc 500 400 500 400TAC control 80%

normalshell devel

Normal devel 70%germination

70% fert.

CHANGES IN COMPANION PROMULGATED METHODS?

• Age of the fish for the acute.Age of the fish for the acute.• Culture and purchase information Culture and purchase information

– more information for more consistencymore information for more consistency– Salinity Tolerance more realisticSalinity Tolerance more realistic

• Analyze results using “all data” Analyze results using “all data” – mean/original regardless of speciesmean/original regardless of species

• All tests must be submitted to regulatory All tests must be submitted to regulatory authority regardless of the outcomeauthority regardless of the outcome

• Compatibility, consistency, greater QA Compatibility, consistency, greater QA required.required.

CLARIFICATION OF PROMULGATED TEST METHODS

• Office of Water memo by Tudor Davies Office of Water memo by Tudor Davies dated 10 April 1996, addresses:dated 10 April 1996, addresses:

»pH and ammonia control»temperature»hardness»test dilution concentrations»acceptance criteria for Champia parvula

ACCURACY AND PRECISION

• AccuracyAccuracy• PrecisionPrecision

» interinterlaboratorylaboratory» intraintralaboratorylaboratory

• Precision estimatesPrecision estimates» Determined as Coefficient of Variation (CV)Determined as Coefficient of Variation (CV)» Statistical measurements of precision: Statistical measurements of precision:

% CV = (standard deviation/mean) X % CV = (standard deviation/mean) X 100100

INTERLABORATORY PRECISION

Ceriodaphnia dubia Chronic Tests Ceriodaphnia dubia Chronic Tests conducted by USEPA Region IV (Athens)conducted by USEPA Region IV (Athens)• Six laboratoriesSix laboratories• Reference toxicant - NaClReference toxicant - NaCl• Endpoint - IC50 and IC25Endpoint - IC50 and IC25

Ceriodaphnia dubiaCV%

Laboratory IC50 IC25

1 19.7 25.82 24.4 24.13 13.6 16.54 28.6 37.65 21.2 28.06 19.3 12.4

CV (%) 21.1 24.1

INTRA- LABORATORY SODIUM CHLORIDE PRECISION DATA

PRECISION: ESTIMATES OF CV’S

Species

AcuteInterlaboratory

ChronicInterlaboratory

C. variegatus 35 - 60 44

P. promelas 12 - 53 27 - 70

M. berylina 42 45

C. dubia 40 – 49 21 - 41

D. magna 51-166 --

INTERLABORATORY PRECISION WEST COAST TESTS

EffluentCoefficient ofVariation (%)

Single ChemicalsCoefficient ofVariation (%)

Test Protocol

Mean Range Mean Range

Holmesimysis 7-dSurvivalGrowth

14 6 – 30 13 1 - 24

M. bahia 4-dSurvival, Acute

65 18-166 41 18 - 61

Mussel LarvalDevelopment

21 2- 42 44 33 – 55

Oyster LarvalDevelopment

29 15 – 41 59 48 – 77

Abalone LarvalDevelopment

14 2 – 25 29 --

Above values were calculated at the lower end of detection

PERFORMANCE EVALUATION STUDIES

USEPA EMSL-Cincinnati Study Number WP026, 1991

Analytes CV (%)

Metals 4.5 - 29.8

Inorganics 6.3 - 36.3

Organics 11.5 - 51.8

DATA EVALUATION• Water quality parameters are taken at beginning and end of Water quality parameters are taken at beginning and end of

each renewal. each renewal.

• Check WQ parameters for within the limitsCheck WQ parameters for within the limits

» DO, Temp., salinity, pH, ammonia, hardness, alkalinityDO, Temp., salinity, pH, ammonia, hardness, alkalinity

• Verify Verify Reference Toxicant testReference Toxicant test was completed and done in was completed and done in comparable/appropriate manner.comparable/appropriate manner.

• Age of the animals, with times and notation on how animals Age of the animals, with times and notation on how animals were obtained or provided.were obtained or provided.

• Summarize and plot the data for Mean/Conc with SD, and Summarize and plot the data for Mean/Conc with SD, and summarize the ranges for replicates; check TAC.summarize the ranges for replicates; check TAC.

• Conduct statistical analysis according to the manual.Conduct statistical analysis according to the manual.

MOST COMMONLY REQUIRED SPECIES IN THE NPDES PERMIT WET PROGRAM

• Ceriodaphnia dubiaCeriodaphnia dubia, waterflea, waterflea

CERIODAPHNIA DUBIA ISSUES• Presence of malesPresence of males

identify under microscopeidentify under microscope>2 per treatment, test suspect>2 per treatment, test suspect

• Test termination - 60% of the animals have Test termination - 60% of the animals have their 3rd brood in controltheir 3rd brood in control identify count live youngidentify count live young

• Reproduction must be Reproduction must be >>15 per surviving 15 per surviving female female and and >>80% survival.80% survival.

Day Count of Young (by replicate)

A B C D E F G H I J

4 0 4 0 0 1 0 2 0 2 0

5 8 0 6 8 4 3 0 2 0 5

6 0 9 3 1 4 7 9 8 9 2

7 13 11 11 12 6 5 2 10 5 9

DISTINGUISHING BROODS FOR THE Ceriodaphnia TEST

MEAN NO. YOUNG: ORIGINAL VS SURVIVING

ConcentrationRep 0 8 15 (rwc) 25 44 100

A 19 10 20 7 D DB 15 25 8 4 D DC 6 17 21 6 D DD 16 19 21 0 D DE 25 11 0 0 D DF 20 12 0/D 0 D DG 13 23 0/D 0/D D DH 10 27 0/D 0/D D DI 12 0/D 0/D 0/D D DJ 0/D 0/D 0/D 0/D D D

Mean Surv. (%) 90 80 50 60 0 0Mean yg/surv. 15.1 18.0 14.0 2.8 0 0Mean yg/orig. 13.6 14.4 7.7 1.7 0 0CV (%) 37.8 36.7 68.3 81.5 0 0

ConcentrationReplicate 0 1 3 10 30 50 75 100

A 23 23 22 19 15 18/D 4/D DB 0/D 24 22 19 8/D 14 0/D DC 28 25 27 M M M 0/D DD 25 20 22 22 21 20/D 0/D DE 27 12 19 19 19 18 0/D DF M M M M M M 0/D DG 22 15 15 17 19 18 0/D DH 24 18 17 17 6 M 0/D DI 22 24 17 17 14 22/D 0/D DJ 26 12/D 18 20 15 20/D 0/D D

Surv 90 90 100 100 90 60 0 0Mean Yg/Surv 24.6 20.1 19.9 18.8 15.6 15.7 7.5 0Mean Yg/Orig 21.9 19.2 19.9 18.8 14.6 15.7 1.5 0CV (%) 38.7 27.1 18.6 9.6 36.3 13.4 66.0 -

1 35246

312546

413652

536124

215364

641352

531624

152463

214653

364215

1 35246

312546

413652

536124

215364

641352

531624

152463

214653

364215

1 0 6 05 04 03 02 0

5 94 93 92 91 99

5 84 83 82 81 88

5 74 73 72 71 77

5 64 63 62 61 66

5 54 53 52 51 55

5 44 43 42 41 44

5 34 33 32 31 33

5 24 23 22 21 22

5 14 13 12 11 11

TAKE HOME POINTS

• Must use approved test methodsMust use approved test methods• Laboratory must follow summary of Laboratory must follow summary of

conditionsconditions• Must meet Test Acceptability Criteria (TAC)Must meet Test Acceptability Criteria (TAC)• Test precision is comparableTest precision is comparable• Selection of laboratory to conduct testing is Selection of laboratory to conduct testing is

critical.critical.

USEPA WET LITIGATION SETTLEMENT

July 24, 1998

• With settlement, comes the “stay of With settlement, comes the “stay of the litigation”the litigation” pending completion of pending completion of tasks by USEPAtasks by USEPA

INTERLABORATORY STUDIES

• USEPA agreed to conduct studies:USEPA agreed to conduct studies: – 3 freshwater chronic 3 freshwater chronic – 4 marine chronic 4 marine chronic – 5 acute tests5 acute tests

• Studies to get underway in Late Winter 99.Studies to get underway in Late Winter 99.• With results, USEPA will ratify or withdraw With results, USEPA will ratify or withdraw

any/all 17 methods and will propose this action any/all 17 methods and will propose this action in September 2000.in September 2000.

METHODOLOGY REQUIREMENTS AND RECOMMENDATIONS

• Modify existing test manualsModify existing test manuals– Clarify mandatory items to assure Clarify mandatory items to assure

consistency in measurementsconsistency in measurements– Clarify discretionary options to Clarify discretionary options to

optimize successful test completion optimize successful test completion

• Rulemaking to convert discretionary provisions: Rulemaking to convert discretionary provisions: – CeriodaphniaCeriodaphnia test randomization and blocking by test randomization and blocking by

parentparent– Development of valid dose concentration response Development of valid dose concentration response

as a pre-requisite for a valid testas a pre-requisite for a valid test– pH control/shiftpH control/shift

• Prepare guidance and recommendations regardingPrepare guidance and recommendations regarding– adjustments to statistical error rate assumptions, adjustments to statistical error rate assumptions,

confidence intervals, available dilution waters, and confidence intervals, available dilution waters, and

permutationspermutations of the valid dose-responseof the valid dose-response

METHODOLOGY REQUIREMENTS AND RECOMMENDATIONS

JANUARY 2001

• Revise manuals to incorporate new Revise manuals to incorporate new requirements and recommendationsrequirements and recommendations

• Current methods provide sufficient direction Current methods provide sufficient direction and flexibility until that time.and flexibility until that time.

ANALYTIC VARIABILITY GUIDANCE

• Recognize variability arises from:Recognize variability arises from:– effluent variabilityeffluent variability– analytic variabilityanalytic variability– analyst variability and moreanalyst variability and more

• Develop guidance on how to take Develop guidance on how to take analytic variability into account for analytic variability into account for determining the need for and derivation determining the need for and derivation of the WET Limit by April 2000.of the WET Limit by April 2000.

You can’t think rationally on an empty stomach

and a whole lot of people can’t do it on a full one either. and a whole lot of people can’t do it on a full one either.

Lord Reith Lord Reith

British administratorBritish administrator

We are inclined to believe those whom we do not know

because they have not deceived us.

Samuel Johnson British lexicographer

Common Name/Species Test DurationSalinityRange Test Endpoint

Topsmelt, Atherinops affinis

7-d 5-36% Growth, Survival

Red abalone, Haliotis rufescens

48-h 36% Larval Development

Mussels, Mytilus spp.,Oyster, Crassostrea gigas

48-h 28-32% Larval Development

Purple urchin, StrongylocentrotuspurpuratusSand dollar, Dendraster excentricus

48-h test, and a1-h test

32-36% Larval Development,

Fertilization

Mysid, Holmesimysis costata 96 h; & 7-d 32-36% Growth, Survival

Giant kelp, Macrocystis pyrifera 48 h, non renew 32-36% Germ-tube Lengthand Germination

WEST COAST MARINE TEST GUIDANCE METHODS

QUALITY ASSURANCE AND QUALITY CONTROL

(QA/QC)

TOPICS TO BE DISCUSSED

• QA/QC procedures within the test QA/QC procedures within the test methods methods

• USEPA DMR/QA ProgramUSEPA DMR/QA Program

• Update of National Environmental Update of National Environmental Laboratory Accreditation Program Laboratory Accreditation Program (NELAP)(NELAP)

TOPICS TO BE DISCUSSED

• Quality AssuranceQuality Assurance

– reliability of datareliability of data

• Quality ControlQuality Control

– procedures within QA procedures within QA programprogram

THE IMPORTANCE OF QUALITY ASSURANCE AND QUALITY CONTROL WHEN CONDUCTING WET TESTS

ELEMENTS OF A QA/QC PROGRAM

• Appoint a QA/QC officeAppoint a QA/QC office• Written QA plan with DQOsWritten QA plan with DQOs• Standard operating procedures (SOP)Standard operating procedures (SOP)• Approved USEPA methods for WETApproved USEPA methods for WET• Record keepingRecord keeping• Qualified staffQualified staff• Suitable space and equipmentSuitable space and equipment• Dilution/culture watersDilution/culture waters

ELEMENTS OF A QA/QC PROGRAM (cont.)

• Effluent sampling and handlingEffluent sampling and handling• Organism health and performanceOrganism health and performance• Test organismsTest organisms• Test conditionsTest conditions• Test acceptability criteriaTest acceptability criteria• Data evaluationData evaluation• Reference toxicantsReference toxicants

IMPORTANCE OF APPOINTING QA OFFICER

• Objectivity in assessing system Objectivity in assessing system performance performance

• Compliance with DQO’sCompliance with DQO’s

INTERNAL AUDITS

DATA QUALITY OBJECTIVES (DQO)

Help ensure that the data collected Help ensure that the data collected are sufficient and of adequate quality are sufficient and of adequate quality for their intended use.for their intended use.

DATA QUALITY CAN BE DESCRIBED BY THE FOLLOWING:

• PrecisionPrecision

• AccuracyAccuracy

• RepresentativenessRepresentativeness

• ComparabilityComparability

• CompletenessCompleteness

Accuracy Precision

WRITTEN DESCRIPTION OF LABORATORY SOPs

RECORD KEEPING AND PREPARATION

QUALIFIED STAFF

LABORATORY SPACE AND EQUIPMENT

INSTRUMENTATION, EQUIPMENT AND SUPPLIES

CONDITIONS OF EQUIPMENT

INSTRUMENT CALIBRATION

LABORATORY WATERSLABORATORY WATERS• Culture waterCulture water

• Diluent reconstituted water - Diluent reconstituted water - – freshwater or salinefreshwater or saline

• Reagent waterReagent water

RECEIVING WATERSRECEIVING WATERS• Uncontaminated waterUncontaminated water

EFFLUENT SAMPLING AND HANDLING

SOURCE AND CONDITION OF ORGANISMS

ORGANISM HEALTH AND PERFORMANCE

TEST CONDITIONS

TEST ACCEPTABILITY CRITERIA (TAC)

PERFORMANCE OF THE CONTROL ORGANISMS

• Acute TACAcute TAC– 90% or greater in control90% or greater in control

• Chronic TACChronic TAC– 80% or greater in control80% or greater in control– growth, reproduction etc.growth, reproduction etc.

DATA ANALYSIS

REFERENCE TOXICANTS

REFERENCE TOXICANT TESTS ARE CONDUCTED TO

DEMONSTATE ONGOING LABORATORY PERFORMANCE

CONTROL CHARTS ARE DEVELOPED FOR EACH TEST

CONDITION AND TEST SPECIES

HOW TO RESPOND TO OUT OF CONTROL REFERENCE

TEST RESULTS

• Check calculations - do they make sense?Check calculations - do they make sense?

• Dilution water quality - does it meet Dilution water quality - does it meet standards?standards?

• QC food preparation and feeding practicesQC food preparation and feeding practices

• Presence of dose response curvePresence of dose response curve

• Glassware cleaning practicesGlassware cleaning practices

CONTROL CHARTS

1 3 5 7 9 11 13 15 17 19

1 3 5 7 9 11 13 15 17 19

A

B

C

Upper Control Limit (x + 2SD)

Central Tendency

Lower Control Limit (x - 2SD)

0 5 10 15

TOXICITY TEST WITH REFERENCE TOXICANTS

LC50

(m

g/L)

789

101112

LC50

(m

g/L)

789

101112

CUMULATIVE TEST NUMBER

20

LC50

TAKE HOME POINTS

• Laboratories must have a QA plan that Laboratories must have a QA plan that addresses all dataaddresses all data

• If QA/QC practices show something that is If QA/QC practices show something that is not correct, corrective actions must take not correct, corrective actions must take placeplace

• Invalid tests must be rerunInvalid tests must be rerun• To keep the laboratory on track, laboratory To keep the laboratory on track, laboratory

management should conduct routine internal management should conduct routine internal auditsaudits

STATES WITH CERTIFICATION PROGRAMS FOR WET OR THAT

INCLUDE WETRegion 1- none

Region 2- New Jersey

Region 3- Virginia

Region 4- N. Carolina- S. Carolina- Florida

Region 5- Wisconsin

Region 6- Arkansas- Louisiana- Oklahoma

Region 7- Iowa- Kansas

Region 8- Utah

Region 9- Arizona- Califonia

Region 10- Washington

DMR QA

DMR QA PURPOSE

A program to evaluate NPDES A program to evaluate NPDES permittee’s ability to analyze and report permittee’s ability to analyze and report accurate self monitoring data.accurate self monitoring data.

PARTICIPATION IS REQUIRED

• Unknown reference toxicant sent to Unknown reference toxicant sent to testing laboratories.testing laboratories.

• Designed to cover as many permittees Designed to cover as many permittees and test conditions as possible.and test conditions as possible.

• Generalized processGeneralized process

PARTICIPATION IS REQUIRED (Cont.)

• Contract laboratories that service Contract laboratories that service multiple permittees need only conduct multiple permittees need only conduct one test for each test type requested.one test for each test type requested.

• WET laboratories receive samples and WET laboratories receive samples and conduct the required tests.conduct the required tests.

PROGRAM REQUIRES BIOLOGICAL, PHYSICAL AND CHEMICAL DATA

REPORTING

PROGRAM REQUIRES BIOLOGICAL, PHYSICAL AND CHEMICAL DATA

REPORTING (Cont.)

• After test completion, laboratory completes report form After test completion, laboratory completes report form and signs certification statement.and signs certification statement.

• Copy sent to EMSL-Cincinnati and NPDES permittee.Copy sent to EMSL-Cincinnati and NPDES permittee.• At USEPA, reported endpoints are statistically At USEPA, reported endpoints are statistically

processed.processed.• Acceptance limits are derived for each test method.Acceptance limits are derived for each test method.• Laboratories falling outside the limits are judged Laboratories falling outside the limits are judged

unacceptable.unacceptable.• Results mailed to participants.Results mailed to participants.

PROGRAM REQUIRES BIOLOGICAL, PHYSICAL AND CHEMICAL DATA

REPORTING (Cont.)

• Follow up on unacceptable results are conducted by Follow up on unacceptable results are conducted by permitting authority.permitting authority.

• Unacceptable results can be due to reporting errors Unacceptable results can be due to reporting errors or analytical errors or both.or analytical errors or both.

• Need corrective actions at support laboratory or by Need corrective actions at support laboratory or by permittee.permittee.

• Need for experienced permitting authority personnel Need for experienced permitting authority personnel to judge the adequacy of the laboratories’ response to judge the adequacy of the laboratories’ response to unacceptable test results.to unacceptable test results.

FUTURE OF THE DMR QA PROGRAM

• May be privatizedMay be privatized

• DownsizedDownsized

• Combination of Combination of solutionssolutions

• Remain the sameRemain the same

STATISTICAL PROCEDURES AND DATA EVALUATION

MODULE OBJECTIVES

• Describe hypothesis testing Describe hypothesis testing proceduresprocedures

• Describe point estimate techniquesDescribe point estimate techniques

• Evaluate acute test resultsEvaluate acute test results

• Evaluate chronic test resultsEvaluate chronic test results

ENDPOINTS

• Biological endpoints:Biological endpoints:

– Lethality, growth, reproduction, etc.Lethality, growth, reproduction, etc.

• Statistical endpoints:Statistical endpoints:

– LC50, NOEC, ICp, etcLC50, NOEC, ICp, etc

STATISTICAL ENDPOINTS

• Acute testsAcute tests

– LC50, EC50, or NOAECLC50, EC50, or NOAEC

• Chronic testsChronic tests

– NOEC, IC25 or EC25NOEC, IC25 or EC25

STATISTICAL APPROACHES

• Hypothesis testingHypothesis testing

– NOECNOEC

• Point estimate techniquesPoint estimate techniques

– LCp, ECp or ICpLCp, ECp or ICp

Purpose of Hypothesis Testsand Basic Considerations

• Purpose - Determine if two things (responses) Purpose - Determine if two things (responses) are differentare different

• Relevance of initial (control) condition(s)Relevance of initial (control) condition(s)• Power of statistical testPower of statistical test

Test #0 2 4 6 8 10 12 14 16

Effe

ct a

t NO

EC

-10

-5

0

5

10

15

20

25

Effects Associated with the NOEC in Fathead Minnow

Growth Data

Purpose of Point-Estimationand Basic Considerations

• Describe relationship between two variables

• Selection of a significant response

•Elucidation of relationship

• Confidence in relationship

0 2 4 6 8 10 120

2

4

6

8

10

12

HYPOTHESIS TESTING FACTS

• NOECs are not point estimatesNOECs are not point estimates• Can’t calculate coefficients of variation or Can’t calculate coefficients of variation or

confidence intervalsconfidence intervals• LOEC may represent a different amount LOEC may represent a different amount

of effect from test to testof effect from test to test• NOEC is a lower concentration level than NOEC is a lower concentration level than

the LOEC when the dose response the LOEC when the dose response curve is smoothcurve is smooth

HYPOTHESIS TESTS

• ParametricParametric

• Non-parametricNon-parametric

PARAMETRIC TESTS

• Must test assumptions:Must test assumptions:– normally distributed datanormally distributed data– tested by Sharpiro-Wilkstested by Sharpiro-Wilks– variance is equalvariance is equal– tested by Bartlett’s testtested by Bartlett’s test

• TestsTests– DunnettsDunnetts– T-test with Bonferroni adjustmentT-test with Bonferroni adjustment

NONPARAMETRIC TESTS

• Nonparametric testsNonparametric tests– based on ranksbased on ranks

• Steel’s Many-one Rank TestSteel’s Many-one Rank Test

• Wilcoxon Rank SumWilcoxon Rank Sum

POINT ESTIMATES FACTS

• Can calculate coefficients of Can calculate coefficients of variation and confidence intervalsvariation and confidence intervals

• Always estimating same effectAlways estimating same effect

• Need specification of a biological Need specification of a biological effect - what value of “p”effect - what value of “p”

METHODS TO CALCULATE LC or EC ENDPOINTS

• ProbitProbit

• Spearman-KarberSpearman-Karber

• Trimmed Spearman-KarberTrimmed Spearman-Karber

• GraphicalGraphical

PROBIT

• AssumptionsAssumptions

• Two partial mortalitiesTwo partial mortalities– chi-square test for heterogeneitychi-square test for heterogeneity

SPEARMAN-KARBER

• A symmetric distribution about the A symmetric distribution about the mean is assumedmean is assumed

• AssumptionsAssumptions– zero response in the lowest toxicant zero response in the lowest toxicant

concentrationconcentration– 100% response in the highest toxicant 100% response in the highest toxicant

concentrationconcentration

INHIBITION CONCENTRATION (ICp)

• Assumptions of ICpAssumptions of ICp

• Responses are monotonically non-Responses are monotonically non-increasingincreasing

BENEFITS OF HYPOTHESIS TESTING

• Results provide information regarding Results provide information regarding test variability (MSD)test variability (MSD)

• Results inform regulator of the no-Results inform regulator of the no-observed effect levelobserved effect level

• The researcher can test just the RWC The researcher can test just the RWC vs. the controlvs. the control

CONCERNS WITH HYPOTHESIS TESTING

• The true effect level may lie The true effect level may lie somewhere in between the NOEC somewhere in between the NOEC and LOECand LOEC

• Poor or excessive statistical powerPoor or excessive statistical power

• Cannot generate intra- or inter-Cannot generate intra- or inter-laboratory precision resultslaboratory precision results

RECCOMMENDATIONS TO HYPOTHESIS TESTING

CONCERNS

• Better spacing of the effluent Better spacing of the effluent concentration - bracket RWCconcentration - bracket RWC

• Establish a test sensitivity criterion-MSD Establish a test sensitivity criterion-MSD limitlimit

MSDThe following formula is used to calculate MSD (as The following formula is used to calculate MSD (as recommended by USEPA 1995)recommended by USEPA 1995)

WhereWhered = critical value for the Dunnett’s procedured = critical value for the Dunnett’s procedure

ssww = square root of the within mean square error (MSE) = square root of the within mean square error (MSE)

nn11 = no. of experimental units in the control treatment = no. of experimental units in the control treatment

n = no. of experimental units per treatment, n = no. of experimental units per treatment, assuming an equal no. in all other treatments.assuming an equal no. in all other treatments.

MSD d sw

(1/n1) (1/n)

BENEFITS OF POINT ESTIMATE TECHNIQUES

• Can quantify intra and inter-laboratory Can quantify intra and inter-laboratory precisionprecision

• Uses all the information from a Uses all the information from a concentration response relationshipconcentration response relationship

• Minimize the importance of the effects Minimize the importance of the effects at the RWCat the RWC

CONCERNS WITH POINT ESTIMATE TECHNIQUES

• Point estimates are model dependent, Point estimates are model dependent, especially for small levels of “p”especially for small levels of “p”

• Appropriate model may vary with Appropriate model may vary with effluent sample, species, amount and effluent sample, species, amount and identity of toxicant(s)identity of toxicant(s)

• Selection of appropriate “p” value to Selection of appropriate “p” value to regulateregulate

EVALUATION OF TOXICITY TESTS

• Verify that appropriate method/species is Verify that appropriate method/species is usedused

• Verify test results meet the required TACVerify test results meet the required TAC• Examine chemical and physical Examine chemical and physical

parameters of testparameters of test• Examine statistical resultsExamine statistical results• Compare test result with permit decision Compare test result with permit decision

criterion limitcriterion limit

The Use of Toxicity Units (TUa/c) to Describe Toxicity

• Acute (TUAcute (TUaa) )

100/LC100/LC5050 or or

(log(100-(log(100-S))/1.7 where S= % S))/1.7 where S= % survival survival in 100 % in 100 % effluent effluent

• Chronic (TUChronic (TUcc))

100/NOEC100/NOECoror 100/(EC100/(ECxx or or

ICICxx))

TUc Calculation Method Comparison

TUc Based Upon NOEC0 1 2 3 4 5 6

TU

c B

ased

Upo

n IC

25

0

1

2

3

4

5

6

IC25 > NOEC

IC25 < NOEC

TAKE HOME POINTS

• The statistical approaches are The statistical approaches are described in the manualsdescribed in the manuals

• Consider additional QA/QC limits for Consider additional QA/QC limits for data (MSD, control CV, etc.) for data (MSD, control CV, etc.) for hypothesis testinghypothesis testing

• Verify that tests meet the required TACVerify that tests meet the required TAC

PERMIT DEVELOPMENT

PERMIT DEVELOPMENTPurpose of Module and Topics to be covered:Purpose of Module and Topics to be covered:• Determine reasonable potential for an excursion Determine reasonable potential for an excursion

above a narrative or numeric WQS.above a narrative or numeric WQS.• Techniques for developing a wasteload allocation Techniques for developing a wasteload allocation

where reasonable potential exists.where reasonable potential exists.• Approaches for devleloping permit limits from the Approaches for devleloping permit limits from the

wastelad allocation.wastelad allocation.• Description of WET requirements in permit limitsDescription of WET requirements in permit limits

INTEGRATED APPROACH TO WQB TOXICS CONTROL

• Chemical-specific ApproachChemical-specific Approach

• Whole Effluent Toxicity ApproachWhole Effluent Toxicity Approach

• Biological Criteria ApproachBiological Criteria Approach

WET CRITERIA

• Narrative - no toxics in toxic Narrative - no toxics in toxic amountsamounts

• Numeric -Numeric -– 10.0 µg/L total copper10.0 µg/L total copper– Acute 0.3 TUa = 100 / LC50 Acute 0.3 TUa = 100 / LC50 – Chronic 1.0 TUc = 100 / NOEC Chronic 1.0 TUc = 100 / NOEC

ACUTE TOXICITY CRITERIA

• Criteria Maximum Concentration Criteria Maximum Concentration (CMC)(CMC)

• Acute 0.3 TUa = 100 / LC50 or Acute 0.3 TUa = 100 / LC50 or EC50EC50

CHRONIC TOXICITY CRITERIA

• Criteria Continuous Concentration (CCC)Criteria Continuous Concentration (CCC)

• Chronic 1.0 TUc = 100 / NOEC or 100 / Chronic 1.0 TUc = 100 / NOEC or 100 / ICpICp

WET NPDES NUMERIC EXAMPLE

• 10.0 µg/L total copper10.0 µg/L total copper

• LC50 50% or TUa = 100 / 50% or 2.0LC50 50% or TUa = 100 / 50% or 2.0

• NOEC 10% or TUc = 100 / 10% or NOEC 10% or TUc = 100 / 10% or 10.010.0

CMC AND CCC CRITERIA APPLY TO MIXING ZONES

• No Allowable Mixing vs. Initial MixingNo Allowable Mixing vs. Initial Mixing– No Allowable Mixing requires the CMC at No Allowable Mixing requires the CMC at

the end-of-pipe and the CCC is determined the end-of-pipe and the CCC is determined by the State.by the State.

– Initial Mixing Zone is an “allocated impact Initial Mixing Zone is an “allocated impact zone” where CMC can’t be exceeded at zone” where CMC can’t be exceeded at edge of the ZID and CCC is determined by edge of the ZID and CCC is determined by the State.the State.

MIXING ZONES

Mixing Mixing ZoneZone

OutfallOutfall

ShorelineShoreline

ZIDZID

C > C M CC > C M C

C C C M C C M C

C > C C CC > C C C

C C C C C C C C

STATE MIXING ZONE CRITERIA

• 9 States:9 States: a specific distancea specific distance

• 29 States:29 States: % of cross-sectional % of cross-sectional area/flowarea/flow

• 4 States: 4 States: a given surface areaa given surface area

• 21 States:21 States: no specific criteriano specific criteria

DETERMINE MIXING ZONES

• Field StudiesField Studies– actual measurement of receiving actual measurement of receiving

water contaminant concentrationswater contaminant concentrations– dye studiesdye studies

• ModelingModeling– calibrated to actual calibrated to actual

observationsobservations– simulate critical conditionssimulate critical conditions

STEPS IN DEVELOPING WET PERMIT LIMITS

• Conduct reasonable potential Conduct reasonable potential analysisanalysis

• Models used in permit developmentModels used in permit development

• Specify all necessary permit Specify all necessary permit conditionsconditions

40 CFR 122.44(d)(1) REGULATORY AUTHORITY

FOR:

• Determining “reasonable potential” to Determining “reasonable potential” to exceed water quality criterion (narrative exceed water quality criterion (narrative or numeric)or numeric)

• Development of permit limits where Development of permit limits where reasonable potential existsreasonable potential exists

FACTORS CONSIDERED IN WET REASONABLE POTENTIAL

EVALUATION (40 CFR 122.44(d)(1)(ii))

• Existing controls on point and nonpoint Existing controls on point and nonpoint sources of pollutionsources of pollution

• Variability of the pollutant or pollutant Variability of the pollutant or pollutant parameter in the effluent parameter in the effluent

• Sensitivity of the species to toxicity testingSensitivity of the species to toxicity testing

• Dilution of the effluent in the receiving waterDilution of the effluent in the receiving water

DETERMINING REASONABLE POTENTIAL WITHOUT EFFLUENT DATA

• Size of discharge / dilution availableSize of discharge / dilution available• History of fish killsHistory of fish kills• Compliance / enforcement historyCompliance / enforcement history• Raw materials usedRaw materials used• Products producedProducts produced• Existence of local pretreatment programExistence of local pretreatment program• Presense of categorical industriesPresense of categorical industries• Chlorination / ammonia problemsChlorination / ammonia problems• Number of commercial and industrial usersNumber of commercial and industrial users

EFFLUENT CHARACTERIZATION FOR REASONABLE POTENTIAL

ANALYSIS• Collect toxicity data in advance of Collect toxicity data in advance of

permit application.permit application.

• USEPA recommends, three species USEPA recommends, three species tested quarterly for a yeartested quarterly for a year

• Conduct toxicity tests based on Conduct toxicity tests based on dilution at edge of mixing zonedilution at edge of mixing zone

REASONABLE POTENTIAL PROCEDURES BASED ON

WET DATA

Must consider uncertainty associated Must consider uncertainty associated with sparse data sets effluent variabilitywith sparse data sets effluent variability

POSSIBLE REASONABLE POTENTIAL OUTCOMES

• Excursions above CMC or CCCExcursions above CMC or CCC– establish WET limitestablish WET limit

• Reasonable potential for excursions above CMC or Reasonable potential for excursions above CMC or CCCCCC– establish WET limitestablish WET limit

• No Reasonable potential for excursion above CMC No Reasonable potential for excursion above CMC or CCCor CCC– conduct WET tests at least once every 5 yearsconduct WET tests at least once every 5 years

• Inadequate informationInadequate information– WET monitoring with reopener clauseWET monitoring with reopener clause

TMDLs and WLAs

• TMDLs required under CWA to TMDLs required under CWA to establish water quality limited stream establish water quality limited stream segments.segments.

• WLAs that portion of TMDLs that WLAs that portion of TMDLs that represent point source dischargesrepresent point source discharges

Pollutant Load

Environmental Conditions

Receiving Water Quality

Reduce Load

Done

M0DEL

Compliance with criteria?

Yes

No

Why Use Models?Models can help determine effluent pollutant

levels that will not violate water quality criteria

WATER QUALITY MODELS

• Used to calculate WLAsUsed to calculate WLAs

• Different types:Different types:– single-value steady-statesingle-value steady-state– two-value steady-statetwo-value steady-state– continuous simulation dynamiccontinuous simulation dynamic– Monte Carlo simulation dynamicMonte Carlo simulation dynamic– lognormal probabilistic dynamiclognormal probabilistic dynamic

WATER QUALITY-BASED PERMIT EFFLUENT GOALS• Account for effluent variabilityAccount for effluent variability

• Achieve water quality protectionAchieve water quality protection– protect WLAprotect WLA– prevent acute and chronic impactsprevent acute and chronic impacts– consider receiving water dilutionconsider receiving water dilution

• AccountabilityAccountability– measure compliancemeasure compliance– be fully enforceablebe fully enforceable

COMPARISON OF WLAs TO PERMIT LIMITS

• WLA:WLA:AcuteAcute 1 hour average1 hour average

ChronicChronic 4 days average4 days average

Human HealthHuman Health Up to 30 days averageUp to 30 days average

• Limit:Limit:MDLMDL Daily maximum limitDaily maximum limit

AMLAML Average monthly limitAverage monthly limit

STATISTICALLY DERIVED PERMIT LIMITS

• AdvantagesAdvantages– reflect effluent variabilityreflect effluent variability– compares acute and chronic WLAs to compares acute and chronic WLAs to

determine the limitsdetermine the limits– factors actual number of samples per month factors actual number of samples per month

into average monthly limit derivationinto average monthly limit derivation

• DisadvantagesDisadvantages– calculations increase burden on permit writerscalculations increase burden on permit writers

ACUTE TO CHRONIC RATIO

• ACR = LC50 / NOEC or 50% / 10%ACR = LC50 / NOEC or 50% / 10%

ACR = 5.0ACR = 5.0

OROR

• ACR = TUc / TUa or 10 / 2ACR = TUc / TUa or 10 / 2

ACR = 5.0ACR = 5.0

DESCRIPTION OF WET PERMIT CONDITIONS

• Test typeTest type• Test species and methodTest species and method• Testing frequencyTesting frequency• Type of sampleType of sample• Selection of dilution water/seriesSelection of dilution water/series• Test duration and typeTest duration and type• Quality assurance proceduresQuality assurance procedures• Statistical endpointStatistical endpoint• Steps to address toxicitySteps to address toxicity

SPECIFICATIONS FOR ACUTE/CHRONIC TOXICITY TESTS

• Acute ToxicityAcute Toxicity– Biological endpoints - lethality or effectBiological endpoints - lethality or effect– Test duration - either 24, 48, or 96 hoursTest duration - either 24, 48, or 96 hours– Test acceptabilityTest acceptability

• Chronic ToxicityChronic Toxicity– Biological endpoints - growth, reproduction, Biological endpoints - growth, reproduction,

fertilization, and lethalityfertilization, and lethality– Test duration - 9 days or lessTest duration - 9 days or less– Test acceptabilityTest acceptability

SELECTION OF TEST METHODS FRESHWATER OR

MARINETYPE OF

RECEIVINGWATER SPECIES

Discharges intofreshwater

Freshwater

Discharges intoestuarine or marinewater body

Marine

SPECIES SELECTION

• Take from approved USEPA test Take from approved USEPA test methodsmethods

• Recommended fish, invertebrate, and Recommended fish, invertebrate, and plantplant

TESTING FREQUENCY

PPPooossssssiiibbbllleee FFFrrreeeqqquuueeennncccyyy ooofffTTTeeessstttiiinnnggg VVVooollluuummmeee ooofff DDDiiisssccchhhaaarrrgggeee

MMMooonnnttthhhlllyyy >>> 111 MMMGGGDDD

QQQuuuaaarrrttteeerrrlllyyy <<< 111 MMMGGGDDD

OOOttthhheeerrr fffaaaccctttooorrrsss tttooo cccooonnnsssiiidddeeerrr::: iiinnnttteeerrrmmmiiitttttteeennnttt dddiiisssccchhhaaarrrgggeee,,,cccooommmpppllliiiaaannnccceee rrreeecccooorrrddd eeeffffffllluuueeennnttt vvvaaarrriiiaaabbbiiillliiitttyyy

SAMPLE COLLECTION

• Type of facilityType of facility

• Effluent variabilityEffluent variability

• Shipping and Shipping and holdingholding

• Special conditionsSpecial conditions

SELECTION OF DILUTION WATER

• Standard laboratory water or receiving Standard laboratory water or receiving waterwater

• Depends on the objectives of the testDepends on the objectives of the test

DILUTION SERIES FOR TESTING

• Multiple dilution concentration testMultiple dilution concentration test

• Single concentration testSingle concentration test

SELECTION OF TEST TYPE

Tests may be conducted as either:Tests may be conducted as either:– static non-renewalstatic non-renewal– static renewalstatic renewal– flow-throughflow-through

QUALITY ASSURANCE PROCEDURES

• Reference Toxicant TestingReference Toxicant Testing– In house culture, monthly testingIn house culture, monthly testing– Outside organisms, concurrent testingOutside organisms, concurrent testing

• Conduct Using the Same Test Conduct Using the Same Test ConditionsConditions

STATISTICAL ENDPOINTS

• AcuteAcute

– LC50LC50

– EC50EC50

– Pass/failPass/fail

• ChronicChronic

– EC/ICpEC/ICp

– NOECNOEC

– Pass/failPass/fail

STEPS TO ADDRESS TOXICITY

• Accelerated TestingAccelerated Testing

• Include TRE/TIE LanguageInclude TRE/TIE Language

COMPLIANCE AND

ENFORCEMENT

NATIONAL WET COMPLIANCE / ENFORCEMENT PERSPECTIVE

USEPA NPDES COMPLIANCE PROGRAM• Self-monitoring approachSelf-monitoring approach

– discharge monitoring report (DMR)discharge monitoring report (DMR)

• Agency inspection approachAgency inspection approach– compliance evaluation inspectioncompliance evaluation inspection– compliance sampling inspectioncompliance sampling inspection– compliance biomonitoring inspectioncompliance biomonitoring inspection– performance audit inspectionperformance audit inspection– pretreatment compliance inspectionpretreatment compliance inspection– toxics diagnostic inspectiontoxics diagnostic inspection

PERFORMANCE AUDIT INSPECTIONS (PAI) LABORATORIES PERFORMING

WET TESTS• Objective is to ensure that a laboratory Objective is to ensure that a laboratory

is providing valid self monitoring data is providing valid self monitoring data consistent with NPDES permit consistent with NPDES permit requirementsrequirements

• Regulatory authorityRegulatory authority– 40 CFR Part 122.41(I)(3)40 CFR Part 122.41(I)(3)– 40 CFR Part 123.2640 CFR Part 123.26– CWA Section 308(a), Part (b)CWA Section 308(a), Part (b)

ELEMENTS OF A PAI

• Regulatory authorityRegulatory authority

• Notification of inspectionNotification of inspection

• Opening conferenceOpening conference

• Records reviewRecords review

• Tour laboratoryTour laboratory

• Closing conference Closing conference

NOTIFICATION OF INSPECTION

• Permittee should contact their contract Permittee should contact their contract laboratorylaboratory

• Accomplished by phone or 308 letterAccomplished by phone or 308 letter

OPENING CONFERENCE

• Credentials presentedCredentials presented

• Right of entry explained, if necessaryRight of entry explained, if necessary

• Purpose and order of inspection Purpose and order of inspection discusseddiscussed

• Only inspecting those test(s) required in Only inspecting those test(s) required in the permitthe permit

RECORDS REVIEW

• Copy of NPDES permit and current Copy of NPDES permit and current method manualsmethod manuals

• All SOP’s and QA/QC manualAll SOP’s and QA/QC manual

• Reference toxicant control chartsReference toxicant control charts

• All logbooks - equipment, organism, etc.All logbooks - equipment, organism, etc.

• Chain-of-custody recordsChain-of-custody records

RECORDS REVIEW (cont.)

• Raw date evaluationRaw date evaluation

• Taxonomic ID of organismsTaxonomic ID of organisms

• Personnel training records and Personnel training records and resumesresumes

• DMR/QA study resultsDMR/QA study results

• Statistical programsStatistical programs

LABORATORY TOUR

• Sample tracking and storage Sample tracking and storage inspectioninspection

• Facility and equipment inspectionFacility and equipment inspection

• Review of test conducted by Review of test conducted by laboratory personnel, if possiblelaboratory personnel, if possible

• Organism culture inspectionOrganism culture inspection

CLOSING CONFERENCE

• Major and minor deficienciesMajor and minor deficiencies

• Recommendations and technical Recommendations and technical assistanceassistance

• General content of final reportGeneral content of final report

COMPLIANCE IS PERMITTEE RESPONSIBILITY

• Compliance is expectedCompliance is expected

• It is the permittee’s responsibility to It is the permittee’s responsibility to achieve complianceachieve compliance

• Permittee should define their own pathPermittee should define their own path

• Permittee should develop and present Permittee should develop and present the means by which compliance will be the means by which compliance will be achievedachieved

USEPA NPDES ENFORCEMENT PROGRAM• Overview of USEPA National Policy on Overview of USEPA National Policy on

WET enforcementWET enforcement

• Common issues raised by the regulated Common issues raised by the regulated communitycommunity

• Examples of USEPA regional Examples of USEPA regional approaches to WET violationsapproaches to WET violations

OVERVIEW OF NATIONAL POLICY AND GUIDANCE

1989 “Whole Effluent Toxicty Basic Permitting 1989 “Whole Effluent Toxicty Basic Permitting Principles and Enforcement Strategy”Principles and Enforcement Strategy”

• Permits must be protective of water quality and Permits must be protective of water quality and written to ensure enforceability.written to ensure enforceability.

• Any violation of a WET limit should be addressed Any violation of a WET limit should be addressed promptly, and it may or may not result in a formal promptly, and it may or may not result in a formal enforcement action.enforcement action.

• A toxicity reduction evaluation (TRE) can be required A toxicity reduction evaluation (TRE) can be required through permit condition or enforcement action.through permit condition or enforcement action.

1989 “ENFORCEMENT MANAGEMENT SYSTEM”

• States that any violation of an NPDES permit is a States that any violation of an NPDES permit is a violation of the CWA for which USEPA encourages violation of the CWA for which USEPA encourages some type of enforcement responsesome type of enforcement response

• Recommends an escalating response to continuing Recommends an escalating response to continuing violations of any parameterviolations of any parameter

• Enforcement responses:Enforcement responses:– infrequent violation causing no known harm results in infrequent violation causing no known harm results in

letter of NOVletter of NOV– known harm or continuing violations results in : AO, APO, known harm or continuing violations results in : AO, APO,

or Civil Referralor Civil Referral

AUGUST 14, 1995 USEPA NATIONAL POLICY MEMORANDUM

• Issued jointly by OWM and ORE to clarify existing Issued jointly by OWM and ORE to clarify existing policy re: single exceedances and inconclusive TREspolicy re: single exceedances and inconclusive TREs

• Section 309 of CWA says any violation of permit limit Section 309 of CWA says any violation of permit limit or condition is subject to enforcement; USEPA’s or condition is subject to enforcement; USEPA’s guidance recommends an escalating response to guidance recommends an escalating response to continuing violationscontinuing violations

• USEPA’s guidance does not recommend that the USEPA’s guidance does not recommend that the initial response to a single exceedance of a WET initial response to a single exceedance of a WET

limit, limit, causing no known harmcausing no known harm, be a, be a formal formal enforcement action with a civil penaltyenforcement action with a civil penalty

AUGUST 14, 1995 USEPA NATIONAL POLICY

MEMORANDUM (cont.)• Section 505 of the CWA allows citizens Section 505 of the CWA allows citizens

to sue anyone alleged “to be in to sue anyone alleged “to be in violation” - case lawviolation” - case law

• OW has established a cooperative OW has established a cooperative agreement with SETAC Foundation to agreement with SETAC Foundation to develop Expert Advisory Panels (EAP)develop Expert Advisory Panels (EAP)

HANDLING WET NON-COMPLIANCE

Treat same as other parametersTreat same as other parameters– treating compliance with WET limits the treating compliance with WET limits the

same as other parameters to the greatest same as other parameters to the greatest extent possible will add the strength of extent possible will add the strength of consistency and precedent to the activityconsistency and precedent to the activity

REGULATORY RELIEF

• How to handle extended How to handle extended periods of noncompliance or periods of noncompliance or known harm?known harm?

• AOs / APOs / Civil ReferralsAOs / APOs / Civil Referrals

• Incentives to solve problemIncentives to solve problem

GUIDE TO CALCULATING CWA SETTLEMENT PENALTIES

• Economic benefitEconomic benefit• Gravity componentsGravity components

– significancesignificance– environmental harm / public healthenvironmental harm / public health– number of violationsnumber of violations– duration of non complianceduration of non compliance

• Adjustment factorAdjustment factor– recalcitrancerecalcitrance– ability to payability to pay– litigation considerationslitigation considerations

PERMITTEE EDUCATION

• Face to face outline of problems / Face to face outline of problems / solutionssolutions

• Written guidanceWritten guidance

• Contractor selectionContractor selection

REGULATORY ACTIVITIES• Initiation of enforcementInitiation of enforcement• Immediate response to non-compliance Immediate response to non-compliance

required, e.g., more frequent testing as a required, e.g., more frequent testing as a prerequisite to the TRE planprerequisite to the TRE plan

• Regulatory follow-up on TRE logicRegulatory follow-up on TRE logic• Periodic update and evaluation of progressPeriodic update and evaluation of progress• Plans must be allowed to workPlans must be allowed to work• Complete follow-through requiredComplete follow-through required• Feedback loopFeedback loop

TOXICITY REDUCTION EVALUATION

TRE APPROACH• What is a TRE and when is it used?What is a TRE and when is it used?• Important elements of a TREImportant elements of a TRE• Logical starting place is a thorough Logical starting place is a thorough

information gathering phaseinformation gathering phase• Preliminary investigation and information Preliminary investigation and information

evaluation of potential causes / sources of evaluation of potential causes / sources of toxicity, effluent variability, treatment toxicity, effluent variability, treatment system efficiency, etc.system efficiency, etc.

TRE REVIEW FOR POTWs

• Accelerated WET testingAccelerated WET testing

• NPDES permit evaluationNPDES permit evaluation– what are the toxicants identified in the permitting what are the toxicants identified in the permitting

process?process?

• Treatment system design criteria, flow Treatment system design criteria, flow diagrams, descriptions of treatment elementsdiagrams, descriptions of treatment elements

• Is the system operating up to design Is the system operating up to design specifications, e.g., removal efficiencies?specifications, e.g., removal efficiencies?

TRE REVIEW FOR POTWs (cont.)

• Updated schematic of sewer collection system.Updated schematic of sewer collection system.• Sources and relative contributions of wastewater?Sources and relative contributions of wastewater?• Influent and effluent flow dataInfluent and effluent flow data• How do flows relate to observed toxicity?How do flows relate to observed toxicity?• Effluent toxicity data, trends, and correlation with Effluent toxicity data, trends, and correlation with

other effluent dataother effluent data• Short or long term variation in toxicity related to Short or long term variation in toxicity related to

flow, rainfall, industrial user operating schedules, flow, rainfall, industrial user operating schedules, seasons.seasons.

TRE REVIEW FOR POTWs (cont.)

• Process control and operational data Process control and operational data and historiesand histories

• In-plant chemical usage (e.g., polymers, In-plant chemical usage (e.g., polymers, coagulants, chlorine)coagulants, chlorine)

• Treatment upset histories and reportsTreatment upset histories and reports

• Pretreatment informationPretreatment information

TRE REVIEW FOR POTWs (cont.)

• Industrial waste surveysIndustrial waste surveys

• Industrial user self-monitoring reportsIndustrial user self-monitoring reports

• Industrial user operational schedules Industrial user operational schedules and flow patternsand flow patterns

• Waste hauler monitoring and Waste hauler monitoring and manifestsmanifests

• Hazardous waste inventoriesHazardous waste inventories

POTW DIFFERENT APPROACHES

• RTAsRTAs– bench simulations of treatment process (activated sludge) bench simulations of treatment process (activated sludge)

to determine toxicity pass-throughto determine toxicity pass-through

• Pretreatment toxicity limitsPretreatment toxicity limits– treatabliltytreatablilty– effects on flow moratoriaeffects on flow moratoria

• Public educationPublic education– environmental and cost considerations of household waste environmental and cost considerations of household waste

disposal practicesdisposal practices– consideration of hazardous waste collection activitiesconsideration of hazardous waste collection activities

TRE REVIEW FOR INDUSTRY

• HousekeepingHousekeeping

• Process and wastewater generating Process and wastewater generating diagrams and descriptionsdiagrams and descriptions

• Diagrams and descriptions of non-Diagrams and descriptions of non-process wastewater sources (cooling process wastewater sources (cooling towers, boilers, floor drains)towers, boilers, floor drains)

• In-plant flow records and water usageIn-plant flow records and water usage

TRE REVIEW FOR INDUSTRY

• Chemical labels, MSDS, and toxicity Chemical labels, MSDS, and toxicity informationinformation

• WWTP operational data and historiesWWTP operational data and histories

• Wastewater monitoring records Wastewater monitoring records (chemical and toxicity)(chemical and toxicity)

• Chemical inventories and usage recordsChemical inventories and usage records

TRE REVIEW FOR INDUSTRY (cont.)

• Chemical treatability informationChemical treatability information

• Production recordsProduction records

• Operating schedules with emphasis on Operating schedules with emphasis on how these schedules affect wastewater how these schedules affect wastewater flow / compositionflow / composition

• Effluent toxicity data and trends and Effluent toxicity data and trends and correlation with other effluent datacorrelation with other effluent data

TRE SUPPORT• Waste reduction / reuse opportunitiesWaste reduction / reuse opportunities

– many states now provide TRE technical many states now provide TRE technical assistance to permitteesassistance to permittees

• Data logsData logs– maintain log of activities, treatment changes, maintain log of activities, treatment changes,

chemical uses, etc.chemical uses, etc.

• Use of innovative TRE approaches, - e.g., Use of innovative TRE approaches, - e.g., toxicity test treatability of process chemicalstoxicity test treatability of process chemicals

TRE REGULATORY ACTION

• Regulatory requirementsRegulatory requirements– advantages / disadvantages from advantages / disadvantages from

permittees perspectivepermittees perspective

• Where to end TRE/TIE?Where to end TRE/TIE?– Is there a need to extend TRE/TIE beyond Is there a need to extend TRE/TIE beyond

the point of compliancethe point of compliance

TOXICITY IDENTIFICATION EVALUATION (TIE)

• CharacterizationCharacterization

• IdentificationIdentification

• ConfirmationConfirmation

CHEMICALS KNOWN TO CAUSE TOXICITY IDENTIFIED FROM PAST

TREs

• Zinc, Nickel, Zinc, Nickel, CopperCopper

• AmmoniaAmmonia

• SurfactantsSurfactants

• ChlorineChlorine

• Salinity (TDS)Salinity (TDS)

• PolymersPolymers

• Diazinon, Diazinon, Chlorofenvinphos, Chlorofenvinphos, Dichlorvos, Dichlorvos, Carbofuran, Carbofuran, MalathionMalathion

• BiocidesBiocides

• Etc.Etc.

CATIONIC METALS AS SUSPECT TOXICANTS(s)

WHEN:• Toxicity reduced by EDTA additionsToxicity reduced by EDTA additions

• Toxicity reduced by sodium Toxicity reduced by sodium thiosulfate additionsthiosulfate additions

• Toxicity reduced by CToxicity reduced by C1818SPE column SPE column

and toxicity no recovered in and toxicity no recovered in methanol eluatemethanol eluate

• Erratic dose responseErratic dose response

CATIONIC METALS AS SUSPECT TOXICANT(s) WHEN

(CONT.):• Graduated pH test show toxicity Graduated pH test show toxicity

differences differences

• Toxicity removed by filtration and Toxicity removed by filtration and recovered in pH3 dilution water extractrecovered in pH3 dilution water extract

• Toxicity removed by cation exchange Toxicity removed by cation exchange resin (e.g., zeolite)resin (e.g., zeolite)

AMMONIA IS SUSPECT TOXICANT WHEN:

• Toxicity reduced with low pH’sToxicity reduced with low pH’s

• Toxicity is greater to fathead minnows Toxicity is greater to fathead minnows than than CeriodaphniaCeriodaphnia

• Post-zeolite column effluent is non-toxic Post-zeolite column effluent is non-toxic and ammonia levels are reducedand ammonia levels are reduced

• Large surface / volume air-stipping Large surface / volume air-stipping reduces toxicityreduces toxicity

• Total ammonia measured is > 5 mg/LTotal ammonia measured is > 5 mg/L

OXIDANTS ARE SUSPECT TOXICANT(s) WHEN:

• Toxicity reduced by sodium thiosulfate additionsToxicity reduced by sodium thiosulfate additions• Toxicity reduced by aerationToxicity reduced by aeration• Toxicity of the sample is less over timeToxicity of the sample is less over time• Ceriodaphnia dubiaCeriodaphnia dubia are more sensitive than are more sensitive than

fathead minnowsfathead minnows• 0.1 mg/L at the effluent LC500.1 mg/L at the effluent LC50

• Toxicity reduced by addition of SOToxicity reduced by addition of SO2-2- saturated saturated

waterwater

PHASE I TIE - CHARACTERIZATION

Chemical / physical manipulations of Chemical / physical manipulations of samples to characterize classes of samples to characterize classes of toxicants, behavior of toxicantstoxicants, behavior of toxicants

– e.g., pH adjustment, aeration, Ce.g., pH adjustment, aeration, C1818

solid phase extraction, filtrationsolid phase extraction, filtration

PHASE II TIE - IDENTIFICATION

• Further analyses to narrow in on specific Further analyses to narrow in on specific toxicant(s), help ID multiple toxicantstoxicant(s), help ID multiple toxicants

• FiltrationFiltration

• SPE fractionsSPE fractions

• Ammonia identification - grad. pH, zeolite, Ammonia identification - grad. pH, zeolite, air strippingair stripping

• Metals identification - EDTA, sodium Metals identification - EDTA, sodium thiosulfate, grad. pHthiosulfate, grad. pH

PHASE III TIE - CONFIRMATION

Confirm that identified toxicant(s) are Confirm that identified toxicant(s) are correctcorrect

– sample spikingsample spiking

– CorrelationCorrelation

– symptomssymptoms

– mass balancemass balance

CHRONIC TIEs

• USEPA has basic methods publishedUSEPA has basic methods published

• More subtle effects than acuteMore subtle effects than acute

• Tiered to make efficient use of timeTiered to make efficient use of time

• Innovative approaches Innovative approaches

– experiments designed ad hoc to experiments designed ad hoc to answer specific questionsanswer specific questions

TREATABILITY

• Cost / benefit decisions between source Cost / benefit decisions between source reduction and treatmentreduction and treatment

• Modifications to existing works / processesModifications to existing works / processes

• Bench / pilot modificationsBench / pilot modifications

• Innovative approachesInnovative approaches– current vs. future availability of processes / current vs. future availability of processes /

products / chemicals and regulatory products / chemicals and regulatory requirementsrequirements

TAKE HOME POINTS

• Handle WET compliance consistent with other limitsHandle WET compliance consistent with other limits– don’t set up a system that will need its own defensedon’t set up a system that will need its own defense

• Compliance is the liability of the permitteeCompliance is the liability of the permittee– don’t assume the responsibility of designing and don’t assume the responsibility of designing and

supporting the TREsupporting the TRE– success or failure is the responsibility of the success or failure is the responsibility of the

permittee and not suggested methodspermittee and not suggested methods– support innovative approachessupport innovative approaches

TAKE HOME POINTS (cont.)

• Emphasize a process with periodic Emphasize a process with periodic feedback and allow it to workfeedback and allow it to work

• Permittee demonstrates its support for Permittee demonstrates its support for the TRE programthe TRE program

SPECIAL TOPICS

SPECIAL WET CONDITIONS

• Inorganic ionic imbalanceInorganic ionic imbalance

• AmmoniaAmmonia

Solutions

• Waste reductionWaste reduction

• More consideration of exposureMore consideration of exposure

• Alternative permitting practicesAlternative permitting practices-- Probabilistic flow permittingProbabilistic flow permitting-- Alternative mixing zonesAlternative mixing zones-- Waste co-minglingWaste co-mingling

Toxicity of Ubiquitous Ions• Generally - Generally -

-- Ca, Mg, Na, KCa, Mg, Na, K-- HCOHCO33, CO, CO33, SO, SO44, Cl, Cl

• OmnipresentOmnipresent• Historically haven’t been controlled as Historically haven’t been controlled as

toxicantstoxicants• Difficult / expensive to treatDifficult / expensive to treat• Osmoregulation problems and concurrent Osmoregulation problems and concurrent

effects on other toxicantseffects on other toxicants

TDS / Salinity / Ultrapure, etc.

• Saline discharges to freshwaterSaline discharges to freshwater-- Pickling, metal plating, textile, ROPickling, metal plating, textile, RO

• Freshwater discharges to marine / estuarineFreshwater discharges to marine / estuarine-- cooling water, POTWscooling water, POTWs

• Ultrapure discharges to freshwater or marineUltrapure discharges to freshwater or marine-- distilled water, groundwaterdistilled water, groundwater

IMBALANCES FROM NATURAL CONDITIONS

pHpH 00 55 1010 1515 2020 25256.56.5 .0261.0261 .0395.0395 .0589.0589 .0865.0865 .125.125 .180.180

7.07.0 .0826.0826 .125.125 .186.186 .273.273 .396.396 .566.566

7.57.5 .261.261 .394.394 .586.586 .859.859 1.241.24 1.771.77

8.08.0 ..820820 1.231.23 1.831.83 2.672.67 3.823.82 5.385.38

8.58.5 2.552.55 3.803.80 5.565.56 7.977.97 11.211.2 15.315.3

9.09.0 7.647.64 11.111.1 15.715.7 21.521.5 28.428.4 36.336.3From 1984 Ambient Water Quality Criteria for AmmoniaFrom 1984 Ambient Water Quality Criteria for Ammonia

Possible Solutions• Permit ammonia as specific chemical to Permit ammonia as specific chemical to

criteriacriteria-- will probably be protective for WET will probably be protective for WET testtest

• Permit WET using probabilistic flow Permit WET using probabilistic flow conditions for winterconditions for winter

• Control pH in test - allowed by USEPA Control pH in test - allowed by USEPA (Davies memo 4/10/96)(Davies memo 4/10/96)

Dilemma• How to write permit WET limits protective How to write permit WET limits protective

of receiving streams during winter?of receiving streams during winter?

• Control pH in the test?Control pH in the test?-- is this indicative of instream / is this indicative of instream /

downstream toxicity?downstream toxicity?

The Ammonia Conundrum• Percent unionized ammonia (toxic form) in Percent unionized ammonia (toxic form) in

a sample increases with increasing pH or a sample increases with increasing pH or temperaturetemperature

• Domestic wastewater tends to increase Domestic wastewater tends to increase pH as it agespH as it ages

• Food presented to chronic tests tends to Food presented to chronic tests tends to increase pH of the solutionincrease pH of the solution

• Chronic tests utilize elevated (25°C) temp.Chronic tests utilize elevated (25°C) temp.

Permitting Case Examples

NORTH CAROLINANORTH CAROLINA

NCAC T15A:02B.0202NCAC T15A:02B.0202 Definitions Definitions

[paraphrasing] [paraphrasing]

(1) Acute Toxicity(1) Acute Toxicity

(a) specific chemicals- one half or less FAV(a) specific chemicals- one half or less FAV

(b) or 1/3 or less lowest LC50(b) or 1/3 or less lowest LC50

(c) effluents- no statistically measurable (c) effluents- no statistically measurable (t-test at 99%CL) (t-test at 99%CL)

(d) or Director may approve case-by-case on (d) or Director may approve case-by-case on conc.- response curve conc.- response curve

NCAC T15A:02B.0202NCAC T15A:02B.0202 Definitions Definitions

• (13) Chronic toxicity to aquatic life means any (13) Chronic toxicity to aquatic life means any harmful effect sustained by either resident harmful effect sustained by either resident aquatic populations or indicator species used as aquatic populations or indicator species used as test organisms in a controlled toxicity test due to test organisms in a controlled toxicity test due to long-term exposure (relative to the life cycle of long-term exposure (relative to the life cycle of the organism) or exposure during a substantial the organism) or exposure during a substantial portion of the duration of a sensitive period of portion of the duration of a sensitive period of the life cycle to a specific chemical substance or the life cycle to a specific chemical substance or mixture of chemicals (as in an effluent).mixture of chemicals (as in an effluent).

NCAC T15A:02B.0208 Standards for Toxic Substances and Temperature

• (a)(1) Aquatic life standards. The concentration of (a)(1) Aquatic life standards. The concentration of toxic substances will not result in chronic toxicity. toxic substances will not result in chronic toxicity. Any levels in excess of the chronic value will be Any levels in excess of the chronic value will be considered to result in chronic toxicity. In the considered to result in chronic toxicity. In the absence of direct measurements of chronic toxicity...absence of direct measurements of chronic toxicity...

[paraphrasing][paraphrasing]• not> prediction of chronic through A:C rationot> prediction of chronic through A:C ratio• not> 1/20 lowest LC50 non-persistentnot> 1/20 lowest LC50 non-persistent• not > 1/100 lowest LC50 persistentnot > 1/100 lowest LC50 persistent

NCAC T15A:02B.0103NCAC T15A:02B.0103 Analytical Procedures Analytical Procedures

• (b) Biological procedures. Biological tests to determine (b) Biological procedures. Biological tests to determine conformity or non-conformity with standards will be based conformity or non-conformity with standards will be based on methods published by the U.S. Environmental on methods published by the U.S. Environmental Protection Agency as outlined in...Protection Agency as outlined in...

...or methods published by the North Carolina Department ...or methods published by the North Carolina Department of Environment, Health and Natural Resources, as of Environment, Health and Natural Resources, as outlined in...outlined in...

...or other such methods as approved as the director...or other such methods as approved as the director

Reasonable PotentialReasonable Potential

• All MajorAll Major• All Complex MinorAll Complex Minor

– Excludes:Excludes:•100% domestic minor•non-contact cooling water

•aquaculture•swimming pools•mine dewatering

• water filtration backwash

• carwashes• rock and gem

mines• sand dredging

LimitsLimits

• IWC>0.25%: Chronic limit at IWC (P/F or P2)IWC>0.25%: Chronic limit at IWC (P/F or P2)• IWC< 0.25: Acute limit at 90% (P/F)IWC< 0.25: Acute limit at 90% (P/F)

• Consideration of diluting flowsConsideration of diluting flows– lakeslakes– tidaltidal

• Consideration of frequency/duration of exposureConsideration of frequency/duration of exposure– stormwaterstormwater– intermittentintermittent

Statistical EndpointsStatistical Endpoints

• AcuteAcute– test of significancetest of significance

• Chronic-Use ChVChronic-Use ChV– Established in regulationEstablished in regulation– Do not have to justify biological Do not have to justify biological

significancesignificance– Have made functional modifications- Have made functional modifications-

PSCPSC

Monitoring FrequencyMonitoring Frequency• Continuous discharge- quarterly in Continuous discharge- quarterly in

specified monthsspecified months– reverts to monthly on single failreverts to monthly on single fail– reverts to quarterly on single passreverts to quarterly on single pass

• Intermittent discharge- more case specificIntermittent discharge- more case specific– first five discharge events then annually first five discharge events then annually

thereafterthereafter– each discharge eventeach discharge event

Species SelectionSpecies Selection

• Chronic-Chronic-Ceriodaphnia dubiaCeriodaphnia dubia

• Acute- Freshwater- Fathead minnowAcute- Freshwater- Fathead minnow

• Acute Saltwater- Acute Saltwater- Mysidopsis bahiaMysidopsis bahia– Saltwater permittees may use a freshwater Saltwater permittees may use a freshwater

species with no salinity modification if species with no salinity modification if desireddesired

• Chronic Saltwater- Chronic Saltwater- Haven’tHaven’t

Practical Sensitivity Criteria Practical Sensitivity Criteria (PSC)(PSC)

• sets lower bound of sets lower bound of C. dubiaC. dubia reproduction reproduction suppression for compliance purposessuppression for compliance purposes

• sets upper bound of maximum statistical sets upper bound of maximum statistical insensitivity for QA purposesinsensitivity for QA purposes

• established by review of thousands of test established by review of thousands of test resultsresults

• reduces disincentive for production of reduces disincentive for production of sensitive datasensitive data

Quality AssuranceQuality Assurance

• Laboratory Certification/ControlLaboratory Certification/Control• Data ReviewData Review

Enforce methodsEnforce methods

Enforce test acceptability criteriaEnforce test acceptability criteria

• Split sample reviewSplit sample review– Data represents reasonable duplication of Data represents reasonable duplication of

analysisanalysis

• DMR/QADMR/QA

Compliance and EnforcementCompliance and Enforcement

• ConsistencyConsistency

• Permittee/Regulator InteractionPermittee/Regulator Interaction

• Permittee’s ResponsibilityPermittee’s Responsibility

• Enforcement/Enforcement ReliefEnforcement/Enforcement Relief

• Complete the ProcessComplete the Process

• Preliminary ActivitiesPreliminary Activities

• In-Depth ActivitiesIn-Depth Activities

Handling WET Non-complianceHandling WET Non-compliance

• Treat same as other parametersTreat same as other parameters– Treating compliance with WET limits the Treating compliance with WET limits the

same as other parameters to the greatest same as other parameters to the greatest extent possible will add the strength of extent possible will add the strength of consistency and precedent to the activityconsistency and precedent to the activity

• Toxic in toxic amounts = non-compliantToxic in toxic amounts = non-compliant

• Toxicity reduced below limit = compliantToxicity reduced below limit = compliant

How to handle extended periods of How to handle extended periods of noncompliance?noncompliance?

• AOs/Consent Orders/DecreesAOs/Consent Orders/Decrees

• Incentives to solve problemIncentives to solve problem

Enforcement/Regulatory Enforcement/Regulatory ReliefRelief

• Face to face outline of problems/solutionsFace to face outline of problems/solutions

• Written GuidanceWritten Guidance

• Contractor SelectionContractor Selection

Permittee EducationPermittee Education

• Establish that the permittee is in non-Establish that the permittee is in non-compliance and that compliance is expectedcompliance and that compliance is expected

• Emphasize it is the permittee’s responsibility Emphasize it is the permittee’s responsibility to achieve complianceto achieve compliance

• Need to let permittee define their own pathNeed to let permittee define their own path• The permittee should develop and present The permittee should develop and present

the means by which compliance will be the means by which compliance will be achievedachieved

Compliance is Permittee Compliance is Permittee ResponsibilityResponsibility

• Initiate enforcementInitiate enforcement• Require immediate response to non-compliance, Require immediate response to non-compliance,

e.g. TRE plan, more frequent testinge.g. TRE plan, more frequent testing• Regulatory follow-up on TRE logicRegulatory follow-up on TRE logic• Periodic update and evaluation of progress reportsPeriodic update and evaluation of progress reports• Allow plans to workAllow plans to work• Requiring complete follow-throughRequiring complete follow-through• Feedback loopFeedback loop

Regulator ActivitiesRegulator Activities

• What is a TRE and when is it used?What is a TRE and when is it used?• Important elements of a TREImportant elements of a TRE• Logical starting place is a thorough Logical starting place is a thorough

information gathering phase. information gathering phase. • Preliminary investigation and information Preliminary investigation and information

evaluation of potential causes/sources of evaluation of potential causes/sources of toxicity, effluent variability, treatment toxicity, effluent variability, treatment system efficiency, etc.system efficiency, etc.

Preliminary TRE ActivitiesPreliminary TRE Activities

• TIETIE

• RTARTA

• Source ReductionSource Reduction

• TreatabilityTreatability

• Discharge AlternativesDischarge Alternatives

Follow-through TRE ActivitiesFollow-through TRE Activities

Handling TIEsHandling TIEs

• Regulatory requirementsRegulatory requirements– Advantages disadvantages from Advantages disadvantages from

permittee’s perspectivepermittee’s perspective

• Where to end TIE?Where to end TIE?– Is there a need to extend TIE and/or TRE Is there a need to extend TIE and/or TRE

beyond the point of compliance?beyond the point of compliance?

Take Home Points - PermittingTake Home Points - Permitting• Establish authority, definitions in regulation. Establish authority, definitions in regulation.

Retain flexibilityRetain flexibility• Apply appropriate exposure considerationsApply appropriate exposure considerations• Write permit language consistent with chemical Write permit language consistent with chemical

specificspecific• Limit the need for site specific considerationLimit the need for site specific consideration• Consider economics of monitoringConsider economics of monitoring• Make it enforceable and enforceMake it enforceable and enforce• Require valid data and enforceRequire valid data and enforce

Take Home Points - Take Home Points - ComplianceCompliance

• Handle compliance consistently with other Handle compliance consistently with other limitslimits

• Compliance is the liability of the permitteeCompliance is the liability of the permittee• Emphasize a process with periodic Emphasize a process with periodic

feedback and allow it to workfeedback and allow it to work• Support a logical, step-wise process of Support a logical, step-wise process of

toxicity reduction that will follow through toxicity reduction that will follow through to successful completionto successful completion