1.1 本指南描述了获取流出物或试验材料（添加到稀释水中，但不添加到食品中）对人体有害影响相关数据的程序 模糊网纹蚤 理查德1894年，在生物体生命的一部分持续暴露期间。这些程序也应该有助于对其他枝角类进行生命周期毒性试验（指南 E1193 )，尽管需要修改。 1.2 这些程序适用于大多数化学品，无论是单独的还是在配方、商业产品或已知混合物中，都可以在水中的必要浓度下准确测量。 经过适当修改，这些程序可用于对温度、溶解氧、pH值、溶解离子和含水废水等材料进行测试（另请参阅指南 E1192 )、浸出液、油、颗粒物、沉积物（另请参阅指南 E1706 )和地表水。更新试验可能不适用于具有高需氧量、高挥发性、快速生物或化学转化或吸附到试验室的材料。如果溶解氧浓度低于4 mg/L或试验材料浓度下降超过20 % 在更新之间任何浓度的测试溶液中，可能需要更频繁的更新。 1.3 根据特殊需要或情况，可能需要对这些程序进行其他修改。使用异常程序进行的测试结果不太可能与许多其他测试的结果具有可比性。比较使用这些程序的修改版本和未修改版本获得的结果，可能会提供有关新概念和程序的有用信息，以便使用 C、 杜比亚。 1.4 本指南安排如下: 部分 参考文件 2. 术语 3. 指南摘要 4. 意义和用途 5. 仪器 6. 设施 6.1 建筑材料 6.2 试验室 6.3 打扫 6.4 试剂和材料 7. 危害 8. 稀释水 9 要求 9.1 来源 9.2 治疗 9.3 刻画 9.4 试验材料 10 全体的 10.1 储备溶液 10.2 流出物 10.3 试验浓度 10.4 收集 10.5 样品容器 10.6 保护 10.7 治疗 10.8 测试生物体 11 种 11.1 年龄 11.2 来源 11.3 亲鱼 11.4 食物 11.5 处理 11.6 质量 11.7 程序 12 可行性论证 12.1 实验设计 12.2 溶解氧 12.3 温度 12.4 准备测试溶液 12.5 调节试验室 12.6 开始测试 12.7 更新测试解决方案 12.8 试验持续时间 12.9 生物学数据 12.10 其他测量 12.11 试验材料 12.12 分析方法学 13 测试的可接受性 14 计算 15 汇报 16 附录 食物 附录X1 培养技术 附录X2 试验室 附录X3 统计指南 附录X4 1.5 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 第节给出了具体的危险说明 8. . 1.6 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 铈中毒 Mount和Norberg首次将其用作毒性试验生物体 ( 2. ) . 引入用于废水和环境水评估， 铈中毒 也成为单一化学测试程序的宝贵补充。 5.2 保护种群需要防止对该物种或试验物种作为替代物种的个体的数量、体重、健康和使用产生不可接受的影响。进行三窝毒性试验，以帮助确定暴露于试验材料后存活率和新生儿数量的变化。 5.3 三次育雏毒性试验的结果 C、 杜比亚 可用于预测在类似条件下暴露对野外物种的慢性或部分慢性影响。 5.4 三个结果- 孵卵毒性试验 C、 杜比亚 可以比较不同物种的慢性敏感性和不同材料的慢性毒性，并研究各种环境因素对此类试验结果的影响。 5.5 三次育雏毒性试验的结果 C、 杜比亚 可能有助于预测同一试验材料在另一水中的同一物种或在同一或不同水中的另一物种的慢性试验结果。大多数这样的预测是基于急性毒性试验的结果，因此三种方法结果的有用性- 孵卵毒性试验 C、 杜比亚 通过报告急性毒性试验的结果，可能会大大增加（见指南 E729 和 E1192 )在相同条件下进行。除了用unfed进行急性试验外 C、 杜比亚 ，也可能需要进行一项急性试验，在该试验中，以与三窝试验中相同的方式喂养生物体，以查看该食物浓度的存在是否会影响急性试验的结果和急性慢性比率（见 10.4.1 ). 5.5.1 有时可以使用已知试验材料从三窝毒性试验中获得48或96小时EC50或LC50，但试验中的所有浓度通常低于EC50或LC50。 此外，在开始三窝试验之前，通常需要了解EC50或LC50，作为确定慢性试验中使用浓度的一种方法（见 10.4.1 ). 应注意的是，由于在慢性试验中添加了食物，急性试验的结果可能不一定与慢性试验的结果一致。 5.6 三次育雏毒性试验 C、 杜比亚 可能有助于研究测试材料的生物可用性和结构-活性关系。 5.7 三次育雏毒性试验的结果 C、 杜比亚 可随温度、食物的质量和数量、溶解离子浓度、稀释水的质量、测试生物体的状况和其他因素而变化。 5.8 三次育雏毒性试验的结果 C、 杜比亚 在评估材料对水生生物的危害时，可能是一个重要的考虑因素（见指南 E1023 )，或在推导水生生物的水质标准时。

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 organism's life. These procedures should also be useful for conducting life cycle toxicity tests with other Cladocera (Guide E1193 ), although modifications will be necessary. 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, dissolved ions, and on such materials as aqueous effluents (see also Guide E1192 ), leachates, oils, particulate matter, sediments (see also Guide E1706 ), 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 4 mg/L or the concentration of test material decreases by more than 20 % in test solution(s) at any concentration 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 guide is arranged as follows: Section Referenced Documents 2 Terminology 3 Summary of Guide 4 Significance and Use 5 Apparatus 6 Facilities 6.1 Construction Materials 6.2 Test Chambers 6.3 Cleaning 6.4 Reagents and Materials 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 Sample Containers 10.6 Preservation 10.7 Treatment 10.8 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 Demonstration of Feasibility 12.1 Experimental Design 12.2 Dissolved Oxygen 12.3 Temperature 12.4 Preparing Test Solutions 12.5 Conditioning Test Chambers 12.6 Beginning a Test 12.7 Renewing Test Solutions 12.8 Duration of Test 12.9 Biological Data 12.10 Other Measurements 12.11 Test Material 12.12 Analytical Methodology 13 Acceptability of Test 14 Calculation 15 Report 16 Appendixes Food Appendix X1 Culture Techniques Appendix X2 Test Chambers Appendix X3 Statistical Guidance Appendix X4 1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements are given in Section 8 . 1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 5.1 Ceriodaphnia was first used as a toxicity test organism by Mount and Norberg ( 2 ) . Introduced for use in effluent and ambient water evaluations, Ceriodaphnia have also been a valuable addition to single chemical test procedures. 5.2 Protection of a population requires prevention of unacceptable effects on the number, weight, health, and uses of the individuals of that species, or species for which the test species serves as a surrogate. A three-brood toxicity test is conducted to help determine changes in survival and the number of neonates produced that result from exposure to the test material. 5.3 Results of three-brood toxicity tests with C. dubia might be used to predict chronic or partial chronic effects on species in field situations as a result of exposure under comparable conditions. 5.4 Results of three-brood toxicity tests with C. dubia might be compared with the chronic sensitivities of different species and the chronic toxicities of different materials, and to study the effects of various environmental factors on results of such tests. 5.5 Results of three-brood toxicity tests with C. dubia might be useful for predicting the results of chronic tests on the same test material with the same species in another water or with another species in the same or a different water. Most such predictions are based on the results of acute toxicity tests, and so the usefulness of the results of a three-brood toxicity test with C. dubia might be greatly increased by also reporting the results of an acute toxicity test (see Guides E729 and E1192 ) conducted under the same conditions. In addition to conducting an acute test with unfed C. dubia , it might also be desirable to conduct an acute test in which the organisms are fed the same as in the three-brood test, to see if the presence of that concentration of that food affects the results of the acute test and the acute chronic ratio (see 10.4.1 ). 5.5.1 A 48 or 96-h EC50 or LC50 can sometimes be obtained from a three-brood toxicity test with a known test material, but often all the concentrations in the test will be below the EC50 or LC50. In addition, it is usually desirable to know the EC50 or LC50 before beginning the three-brood test, as a means to determine the concentrations for use in the chronic test (see 10.4.1 ). It should be noted that results from an acute test may not necessarily correspond to those of a chronic test, due to the addition of food to the chronic test. 5.6 Three-brood toxicity tests with C. dubia might be useful for studying biological availability of, and structure activity relationships between, test materials. 5.7 Results of three-brood toxicity tests with C. dubia can vary with temperature, quality and quantity of food, dissolved ion concentrations, quality of the dilution water, condition of the test organisms, and other factors. 5.8 Results of three-brood toxicity tests with C. dubia might be an important consideration when assessing the hazards of materials to aquatic organisms (see Guide E1023 ), or when deriving water quality criteria for aquatic organisms.