Short-term Methods for Estimating the Chronic Toxicity of

5,89
MB Effluents and Receiving Waters to Freshwater Organisms

350
stron

5075
ID UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

2002
rok

CONTENTS

Figures vii

Tables . x

Section Number Page

1. Introduction . 1

2. Short-Term Methods for Estimating Chronic Toxicity 3

Introduction 3

Types of Tests 5

Static Tests 5

Advantages and Disadvantages of Toxicity Test Types 6

3. Health and Safety . 7

General Precautions 7

Safety Equipment . 7

General Laboratory and Field Operations . 7

Disease Prevention 8

Safety Manuals . 8

Waste Disposal . 8

4. Quality Assurance 9

Introduction 9

Facilities, Equipment, and Test Chambers 9

Test Organisms . 9

Laboratory Water Used for Culturing and Test Dilution Water 9

Effluent and Receiving Water Sampling and Handling 10

Test Conditions . 10

Quality of Test Organisms 10

Food Quality . 11

Acceptability of Short-Term Chronic Toxicity Tests 11

Analytical Methods 11

Calibration and Standardization 12

Replication and Test Sensitivity 12

Variability in Toxicity Test Results . 12

Test Precision 12

Demonstrating Acceptable Laboratory Performance 15

Documenting Ongoing Laboratory Performance . 15

Reference Toxicants . 16

Record Keeping . 18

5. Facilities, Equipment, and Supplies . 19

General Requirements 19

Test Chambers 20

Cleaning Test Chambers and Laboratory Apparatus 20

Apparatus and Equipment for Culturing and Toxicity Tests 21

Reagents and Consumable Materials 21

Test Organisms . 21

Supplies . 21

6. Test Organisms . 22

Test Species 22

Sources of Test Organisms 22

Life Stage . 23

Laboratory Culturing . 24

Holding and Handling Test Organisms . 24

Transportation to the Test Site . 24

Test Organism Disposal 25

7. Dilution Water . 26

Types of Dilution Water 26

Standard, Synthetic Dilution Water . 26

Use of Receiving Water as Dilution Water 28

Use of Tap Water as Dilution Water . 29

Dilution Water Holding . 29

8. Effluent and Receiving Water Sampling, Sample Handling, and Sample Preparation for Toxicity

Tests . 30

Effluent Sampling . 30

Effluent Sample Types . 30

Effluent Sampling Recommendations 31

Receiving Water Sampling 31

Effluent and Receiving Water Sample Handling, Preservation, and Shipping . 32

Sample Receiving . 33

Persistence of Effluent Toxicity During Sample Shipment and Holding . 33

Preparation of Effluent and Receiving Water Samples for Toxicity Tests 33

Preliminary Toxicity Range-Finding Tests 35

Multi-concentration (Definitive) Effluent Toxicity Tests . 36

Receiving Water Tests . 36

9. Chronic Toxicity Test Endpoints and Data Analysis 37

Endpoints 37

Relationship Between Endpoints Determined by

Hypothesis Testing and Point Estimation Techniques 37

Precision 39

Data Analysis 39

Choice of Analysis 41

Hypothesis Tests 44

Point Estimation Techniques . 45

10. Report Preparation and Test Review 47

Report Preparation . 47

Test Review 49

11. Test Method: Fathead Minnow, Pimephales promelas, Larval Survival and Growth Test Method

1000.0 53

Scope and Application 53

Summary of Method 53

Interferences 53

Safety 58

Apparatus and Equipment 58

Reagents and Consumable Materials . 59

Effluent and Receiving Water Collection, Preservation, and Storage 65

Calibration and Standardization . 65

Quality Control 65

Test Procedures 65

Summary of Test Conditions and Test Acceptability Criteria 72

Acceptability of Test Results . 77

Data Analysis . 77

Precision and Accuracy 106

12. Test Method: Fathead Minnow, Pimephales promelas, Embryo-larval Survival and Teratogenicity

Test Method 1001.0 . 112

Scope and Application 112

Summary of Method 112

Interferences 112

Safety 114

Apparatus and Equipment 114

Reagents and Consumable Materials . 115

Effluent and Receiving Water Collection, Preservation, and Storage 116

Calibration and Standardization . 116

Quality Control 117

Test Procedures 117

Summary of Test Conditions and Test Acceptability Criteria 124

Acceptability of Test Result 124

Data Analysis . 128

Precision and Accuracy 136

13. Test Method: Daphnid, Ceriodaphnia dubia, Survival and Reproduction Test Method 1002.0 141

Scope and Application 141

Summary of Method 141

Interferences 141

Safety 143

Apparatus and Equipment 143

Reagents and Consumable Materials . 145

Effluent and Receiving Water Collection, Preservation, and Storage 154

Calibration and Standardization . 154

Quality Control 154

Test Procedures 154

Summary of Test Conditions and Test Acceptability Criteria 161

Acceptability of Test Results . 161

Data Analysis . 166

Precision and Accuracy 189

14. Test Method: Green Alga, Selenastrum capricornutum, Growth

Test Method 1003.0 197

Scope and Application 197

Summary of Method 197

Interferences 197

Safety 197

Apparatus and Equipment 198

Reagents and Consumable Materials . 199

Effluent and Receiving Water Collection, Preservation, and Storage 203

Calibration and Standardization . 203

Quality Control 203

Test Procedures 203

Summary of Test Conditions and Test Acceptability Criteria 208

Acceptability of Test Results . 208

Data Analysis . 212

Precision and Accuracy 224