| | Short-term Methods for Estimating the Chronic Toxicity of |
| | 10,3 | | MB | Effluents and Receiving Waters to Marine and Estuarine |
| | 486 | | stron | Organisms |
| | 5076 | | ID | UNITED STATES ENVIRONMENTAL PROTECTION AGENCY |
| | 2002 | | rok |
| | CONTENTS |
| | Figures . viii |
| | Tables xiii |
| | Section Number Page |
| | 1. Introduction . 1 |
| | 2. Short-Term Methods for Estimating Chronic Toxicity 3 |
| | Introduction . 3 |
| | Types of Tests 5 |
| | Static Tests 6 |
| | 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 Oranisms 9 |
| | Laboratory Water Used for Culturing and 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 Chronic Toxicity Tests . 11 |
| | Analytical Methods 12 |
| | 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 . 20 |
| | Reagents and Consumable Materials . 20 |
| | Test Organisms 21 |
| | Supplies 21 |
| | 6. Test Organisms . 22 |
| | Test Species . 22 |
| | Sources of Test Organisms . 22 |
| | Life Stage 23 |
| | Laboratory Culturing . 23 |
| | 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 . 30 |
| | 8. Effluent and Receiving Water Sampling, Sample Handling,and Sample Preparation for |
| | Toxicity Tests 31 |
| | Effluent Sampling 31 |
| | Effluent Sample Types 31 |
| | Effluent Sampling Recommendations 32 |
| | Receiving Water Sampling . 32 |
| | Effluent and Receiving Water Sample Handling, Preservation, and Shipping . 33 |
| | Sample Receiving 34 |
| | Persistence of Effluent Toxicity During Sample Shipment and Holding 34 |
| | Preparation of Effluent and Receiving Water Samples for Toxicity Tests . 34 |
| | Preliminary Toxicity Range-finding Tests . 38 |
| | Multiconcentration (Definitive) Effluent Toxicity Tests 38 |
| | Receiving Water Tests 38 |
| | 9. Chronic Toxicity Test Endpoints and Data Analysis 40 |
| | Endpoints 40 |
| | Relationship between Endpoints Determined by Hypothesis Testing and Point Estimation |
| | Techniques 40 |
| | Precision . 42 |
| | Data Analysis . 42 |
| | Choice of Analysis . 44 |
| | Hypothesis Tests . 46 |
| | Point Estimation Techniques . 47 |
| | 10. Report Preparation and Test Review 49 |
| | Report Preparation 49 |
| | Test Review 51 |
| | 11. Test Method: Sheepshead Minnow, Cyprinodon variegatus, Larval Survival and GrowthTest |
| | Method 1004.0 . 55 |
| | Scope and Application . 55 |
| | Summary of Method 55 |
| | Interferences 55 |
| | Safety 57 |
| | Apparatus and Equipment 57 |
| | Reagents and Consumable Materials 59 |
| | Effluent and Receiving Water Collection, Preservation, and Storage . 67 |
| | Calibration and Standardization . 67 |
| | Quality Control 67 |
| | Test Procedures 67 |
| | Summary of Test Conditions and Test Acceptability Criteria . 76 |
| | Acceptability of Test Results 76 |
| | Data Analysis 76 |
| | Precision and Accuracy . 107 |
| | 12. Test Method: Sheepshead Minnow, Cyprinodon variegatus, Embryo-larval Survival and |
| | Teratogenicity Test Method 1005.0 . 117 |
| | Scope and Application 117 |
| | Summary of Method . 117 |
| | Interferences . 117 |
| | Safety . 119 |
| | Apparatus and Equipment . 119 |
| | Reagents and Consumable Materials . 121 |
| | Effluent and Receiving Water Collection, Preservation, and Storage 130 |
| | Calibration and Standardization 130 |
| | Quality Control . 130 |
| | Test Procedures . 130 |
| | Acceptability of Test Results . 136 |
| | Summary of Test Conditions and Test Acceptability Criteria 136 |
| | Data Analysis . 136 |
| | Precision and Accuracy . 152 |
| | 13. Test Method: Inland Silverside, Menidia beryllina, Larval |
| | Survival and Growth Method 1006.0 . 155 |
| | Scope and Application 155 |
| | Summary of Method . 155 |
| | Interferences . 155 |
| | Safety . 157 |
| | Apparatus and Equipment . 157 |
| | Reagents and Consumable Materials . 159 |
| | Effluent and Receiving Water Collection, Preservation, and Storage 168 |
| | Calibration and Standardization 168 |
| | Quality Control . 168 |
| | Test Procedures . 168 |
| | Summary of Test Conditions and Test Acceptability Criteria 175 |
| | Acceptability of Test Results . 175 |
| | Data Analysis . 175 |
| | Precision and Accuracy . 205 |
| | 14. Test Method: Mysid, Mysidopsis bahia, Survival, Growth, and Fecundity Test Method 1007.0 |
| | 214 |
| | Scope and Application 214 |
| | Summary of Method . 214 |
| | Interferences . 214 |
| | Safety . 216 |
| | Apparatus and Equipment . 216 |
| | Reagents and Consumable Materials . 217 |
| | Effluent and Receiving Water Collection, Preservation, and Storage 226 |
| | Calibration and Standardization 226 |
| | Quality Control . 226 |
| | Test Procedures . 227 |
| | Summary of Test Conditions and Test Acceptability Criteria 235 |
| | Acceptability of Test Results . 235 |
| | Data Analysis . 236 |