1. Scope 1.1 This guide describes procedures for obtaining data concerning the adverse effects of an effluent or a test material (added to dilution water, but not to food) on Ceriodaphnia dubia Richard 1894, during continuous exposure throughout a portion of the organisms life. These procedures should also be useful for conducting life cycle toxicity tests with other Cladocera (Guide E1193), although modifications will be necessary. Tests of effluents present particular challenges relative to dilution water, acclimation, etc. 1.2 These procedures are applicable to most 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, pH, and on such materials as aqueous effluents (see also Guide E1192), leachates, oils, particulate matter, sediments, and surface waters. Renewal tests might not be applicable to materials that have high oxygen demand, are highly volatile, are rapidly biologically or chemically transformed, or sorb to test chambers. If the concentration of dissolved oxygen falls below 40% of saturation or the concentration of test material decreases by more than 20% in test solution(s) between renewals, more frequent renewals might be necessary. 1.3 Other modifications of these procedures might be justified by special needs or circumstances. Results of tests conducted using unusual procedures are not likely to be comparable to results of many other tests. Comparisons of results obtained using modified and unmodified versions of these procedures might provide useful information on new concepts and procedures for conducting three brood toxicity tests with C. dubia. 1.4 This standard may involve hazardous materials, operations, and equipment. This standard does not purport to address all of the safety problems 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 8. 1.5 This guide is arranged as follows: Section Referenced Documents 2 Terminology 3 Summary of Standard 4 Significance and Use 5 Apparatus 6 Facilities 6.1 Construction Materials 6.2 Test Chamber 6.3 Cleaning 6.4 Demonstration of Feasibility 6.5 Reagents 7 Hazards 8 Dilution water 9 Requirements 9.1 Source 9.2 Treatment 9.3 Characterization 9.4 Test Material 10 General 10.1 Stock Solution 10.2 Effluent 10.3 Test Concentration(s) 10.4 Collection 10.5 Preservation 10.6 Treatment 10.7 Test Organisms 11 Species 11.1 Age 11.2 Source 11.3 Brood Stock 11.4 Food 11.5 Handling 11.6 Quality 11.7 Procedure 12 Experimental Design 12.1 Dissolved Oxygen 12.2 Temperature 12.3 Preparing Test Solutions 12.4 Conditioning Test Chambers 12.5 Beginning the Test 12.6 Renewing Test Solutions 12.7 Duration of Test 12.8 Biological Data 12.9 Other Measurements 12.10 Analytical Methodology 13 Acceptability of Test 14 Calculation 15 Report 16 Appendices Food X1 Culture Techniques X2 Test Chambers X3 Statistical Guidance X4