| | Methods for Measuring the Acute Toxicity of Effluents and |
| | 2,98 | | MB | Receiving Waters to Freshwater and Marine Organisms |
| | 275 | | stron |
| | 5074 | | ID | UNITED STATES ENVIRONMENTAL PROTECTION AGENCY |
| | 2002 | | rok |
| | CONTENTS |
| | Figures vi |
| | Tables vii |
| | 1. Introduction . 1 |
| | 2. Types of Tests . 2 |
| | 3. Health and Safety . 5 |
| | General Precautions . 5 |
| | Safety Equipment . 5 |
| | General Laboratory and Field Operation . 5 |
| | Disease Prevention 6 |
| | Safety Manuals . 6 |
| | Waste Disposal . 6 |
| | 4. Quality Assurance 7 |
| | Introduction . 7 |
| | Facilities, Equipment, and Test Chambers 7 |
| | Test Organisms . 7 |
| | Laboratory Water Used for Culturing and Test Dilution Water . 7 |
| | Effluent Sampling and Sample Handling . 8 |
| | Test Conditions . 8 |
| | Quality of Test Organisms 8 |
| | Food Quality . 8 |
| | Acceptability of Acute Toxicity Test Results . 9 |
| | Analytical Methods . 9 |
| | Calibration and Standardization 9 |
| | Replication and Test Sensitivity . 10 |
| | Variability in Toxicity Test Results 10 |
| | Demonstrating Acceptable Laboratory Performance . 18 |
| | Documenting Ongoing Laboratory Performance 18 |
| | Reference Toxicants 21 |
| | Record Keeping . 21 |
| | 5. Facilities and Equipment 22 |
| | General Requirements 22 |
| | Cleaning Test Chambers and Laboratory Apparatus . 23 |
| | Apparatus and Equipment for Culturing and Toxicity Tests 23 |
| | Reagents and Consumable Materials . 24 |
| | Test Organisms 26 |
| | 6. Test Organisms . 27 |
| | Test Species 27 |
| | Sources of Test Organisms . 28 |
| | Life Stage 29 |
| | Laboratory Culturing . 29 |
| | Holding and Handling Test Organisms . 29 |
| | Transportation to the Test Site 29 |
| | Test Organism Disposal . 30 |
| | 7. Dilution Water 31 |
| | Types of Dilution Water . 31 |
| | Standard, Synthetic Dilution Water 31 |
| | Use of Receiving Water as Dilution Water 33 |
| | Use of Tap Water as Dilution Water . 35 |
| | Dilution Water Holding . 36 |
| | 8. Effluent and Receiving Water Sampling and Sample Handling 37 |
| | Effluent Sampling 37 |
| | Effluent Sample Types 37 |
| | Effluent Sampling Recommendations 38 |
| | Receiving Water Sampling . 39 |
| | Effluent and Receiving Water Sample Handling, Preservation, and Shipping . 39 |
| | Sample Receiving 40 |
| | Persistence of Effluent Toxicity During Sample Shipment and Holding . 40 |
| | 9. Acute Toxicity Test Procedures 41 |
| | Preparation of Effluent and Receiving Water Samples for Toxicity Tests 41 |
| | Preliminary Toxicity Range-finding Tests . 43 |
| | Multi-Concentration (Definitive) Effluent Toxicity Tests . 43 |
| | Receiving Water Tests 43 |
| | Static Tests . 44 |
| | Flow-Through Tests 45 |
| | Number of Test Organisms 45 |
| | Replicate Test Chambers 45 |
| | Loading of Test Organisms 46 |
| | Illumination 46 |
| | Feeding 46 |
| | Test Temperature 46 |
| | Stress 47 |
| | Dissolved Oxygen Concentration 47 |
| | Test Duration . 50 |
| | Acceptability of Test Results . 50 |
| | Summary of Test Conditions for the Principal Test Organisms 50 |
| | 10. Test Data 67 |
| | Biological Data 67 |
| | Chemical and Physical Data 67 |
| | 11. Acute Toxicity Data Analysis 71 |
| | Introduction 71 |
| | Determination of the LC50 from Definitive, Multi-Effluent-Concentration Acute Toxicity Tests 72 |
| | The Graphical Method 72 |
| | The Spearman-Karber Method 76 |
| | The Trimmed Spearman-Karber Method 78 |
| | The Probit Method . 81 |
| | Determination of No-Observed-Adverse-Effect Concentration (NOAEC) from Multi-Concentration |
| | Tests, and Determination of Pass or Fail (Pass/Fail) for Single-Concentration (Paired) Tests 83 |
| | General Procedure 88 |
| | Single Concentration Test 95 |
| | Multi-Concentration Test 98 |
| | 12. Report Preparation and Test Review 109 |
| | Report Preparation 109 |
| | Introduction 109 |
| | Plant Operations 109 |
| | Source of Effluent, Receiving Water, and Dilution Water 109 |
| | Test Conditions . 110 |
| | Test Organisms . 110 |
| | Quality Assurance . 110 |
| | Results 110 |
| | Conclusions and Recommendations . 110 |
| | Test Review . 111 |
| | Sampling and Handling . 111 |
| | Test Acceptability Criteria 111 |
| | Test Conditions . 111 |
| | Statistical Methods 111 |
| | Concentration-Response Relationships . 112 |
| | Reference Toxicant Testing . 112 |
| | Test Variability . 113 |
| | Cited References 114 |
| | Bibliography . 119 |
| | Appendices . 124 |
| | A. Distribution, Life Cycle, Taxonomy, and Culture Methods 125 |
| | A.1. Ceriodaphnia dubia 125 |
| | A.2. Daphnia (D. magna and D. pulex) 140 |
| | A.3. Mysids (Mysidopsis bahia and Holmesimysis costata) . 159 |
| | A.4. Brine Shrimp (Artemia salina) . 178 |
| | A.5. Fathead Minnow (Pimephales promelas) . 185 |
| | A.6. Rainbow Trout, Oncorhynchus mykiss and Brook Trout, Salvelinus fontinalis 201 |
| | A.7. Sheepshead minnow (Cyprinodon variegatus) . 209 |
| | A.8. Silversides: Inland Silverside (Mendia beryllina), Atlantic Silverside (M. menidia), and Tidewater |
| | Silverside (M. peninsulae) . 224 |
| | B. Supplemental List of Acute Toxicity Test Species . 238 |
| | C. Dilutor Systems 240 |
| | D. Plans for Mobile Toxicity Test Laboratory . 253 |
| | D.1. Tandem-Axle Trailer . 253 |
| | D.2. Fifth Wheel Trailer 256 |
| | E. Check Lists and Information Sheets 257 |
| | E.1. Toxicity Test Field Equipment List . 257 |
| | E.2. Information Check List for On-Site Industrial or Municipal Toxicity Test . 259 |
| | E.3. Daily Events Log 264 |
| | E.4. Dilutor Calibration Form 265 |
| | E.5. Daily Dilutor Calibration Check 266 |