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1.1 This guide describes procedures for obtaining laboratory data concerning the adverse effects of a test material added to dilution water-but not to food-on certain species of freshwater and saltwater fishes during 28 to 120-day (depending on species) continuous exposure, beginning before hatch, and ending after hatch, using the flow-through technique. This guide will probably be useful for conducting early life-stage toxicity tests with some other species of fish, although modifications might be necessary.

1.2 Other modifications of these procedures might be justified by special needs or circumstances. Although using appropriate procedures is more important than following prescribed procedures, results of tests conducted using unusual procedures are not likely to be comparable to results of many other tests. Comparison of results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting early life-stage toxicity tests with fishes.

1.3 These procedures are applicable to all chemicals, either individually or in formulations, commercial products, or known mixtures, that can be measured accurately at the necessary concentrations in water. With appropriate modifications these procedures can be used to conduct tests on temperature, dissolved oxygen, and pH and on such materials as aqueous effluents (see Guide E1192), leachates, oils, particulate matter, sediments, and surface waters.

1.4 This guide is arranged as follows:

Section Referenced Documents 2 Terminology 3 Summary of Standard 4 Significance and Use 5 Hazards 6 Apparatus 7 Facilities 7.1 Construction Materials 7.2 Metering System 7.3 Test Chambers and Incubation Cups 7.4 Cleaning 7.5 Acceptability 7.6 Dilution Water 8 Requirements 8.1 Source 8.2 Treatment 8.3 Characterization 8.4 Test Material 9 General 9.1 Stock Solution 9.2 Test Concentration(s) 9.3 Test Organisms 10 Species 10.1 Age 10.2 Source 10.3 Brood Stock 10.4 Handling 10.5 Procedure 11 Experimental Design 11.1 Dissolved Oxygen 11.2 Temperature 11.3 Beginning the Test 11.4 Thinning 11.5 Feeding 11.6 Duration of Test 11.7 Biological Data 11.8 Other Measurements 11.9 Analytical Methodology 12 Acceptability of Test 13 Calculation of Results 14 Documentation 15 Appendixes X1. Salmon, Trout, and Char X2. Northern pike X3. Fathead minnow X4. White sucker X5. Channel catfish X6. Bluegill X7. Gulf toadfish X8. Sheepshead minnow X9. Silversides X10. Statistical Guidance X11. Striped Bass

1.5 This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements are given in Section 6 and 9.