1.1 本指南涵盖了获取实验室数据的程序，这些数据涉及水性流出物对某些淡水和咸水鱼类、大型无脊椎动物和两栖动物的不利影响，通常在2天至4天内 根据物种，使用静态、更新和流通技术进行日间暴露。这些程序可能有助于对许多其他水生物种的废水进行急性毒性试验，尽管可能需要进行修改。 1.2 这些程序的其他修改可能因特殊需要或情况而合理。尽管使用适当的程序比遵循规定的程序更重要，但使用非常规程序进行的测试结果不太可能与许多其他测试结果相比较。 使用这些程序的修改版本和未修改版本获得的结果的比较可能会提供有关对水性流出物进行急性毒性试验的新概念和程序的有用信息。 1.3 本指南主要基于指南 E729型 其中提供了适用于本方法中描述的环境和流出物毒性测试的测试元件的附加细节。两种指南的主要区别在于( 1. )最大测试浓度为100 % 流出物或环境样品( 2. )测试不是化学特定的，并且( 3. )流出物和环境样品的保持时间通常比化学品和其他测试材料的保持时间短得多。由于样品通常是一种复杂的化学混合物，分析测试通常不能用于确认暴露浓度。 1.4 在特定情况下使用的技术的选择将取决于研究者的需求和可用资源。静态测试提供了最容易获得的急性毒性测量，但不应持续48小时。更新和流通测试可能持续48小时以上，因为溶解氧和流出物的pH和浓度保持在所需水平，降解和代谢产物被去除。静态试验可能不适用于含氧量高或含有以下物质的废水( 1. )高度挥发性( 2. )在水溶液中快速生物或化学转化，或( 3. )在测试过程中，测试室或生物体从测试溶液中大量移除。 流式测试通常优于更新测试，尽管在某些情况下，更新测试可能比流式测试更具成本效益。 1.5 在制定这些程序时，试图平衡科学和实际考虑，并确保结果对于通常使用的应用而言足够准确和精确。一个主要考虑因素是，废水急性试验结果的一般用途不需要或不需要比本指南中规定的要求更严格的要求。尽管通过使用更多的生物、更长的适应时间等可以改进测试，但本指南中提出的要求通常是足够的。 1.6 急性毒性试验的结果通常应以半数致死浓度（LC50）或半数有效浓度（EC50）报告。 在某些情况下，可能只需要确定特定浓度对测试物种是否具有急性毒性，或者LC50或EC50是否高于或低于特定浓度。 1.7 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. 危害 7. 仪器 6. 设施 6.1 特殊要求 6.2 建筑材料 6.3 计量系统 6.4 试验室 6.5 打扫 6.6 可接受性 6.7 稀释水 8. 要求 8.1 来源 8.2 治疗 8.3 刻画 8.4 流出物 9 采样点 9.1 收集 9.2 保护 9.3 治疗 9.4 试验浓度（s） 9.5 测试生物体 10 种 10.1 年龄 10.2 来源 10.3 保养和处理 10.4 喂养 10.5 疾病治疗 10.6 霍尔丁 10 7. 适应环境 10.8 质量 10.9 程序 11 实验设计 11.1 溶解氧 11.2 温度 11.3 加载 11.4 开始测试 11.5 喂养 11.6 试验持续时间 11.7 生物学数据 11.8 其他测量值 11.9 分析方法学 12 测试的可接受性 13 计算或结果 14 汇报 15 1.8 本标准并不旨在解决与其使用相关的所有安全问题（如有）。本标准的使用者有责任在使用前建立适当的安全、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害说明 7. . 1.9 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《国际标准、指南和建议制定原则决定》中确立的国际公认标准化原则制定的。 =====意义和用途====== 5.1 进行急性流出物毒性试验，以获得有关在特定实验条件下短期暴露于流出物对试验生物的直接影响的信息。人们可以直接检查废水和一些环境水中出现的复杂化学混合物的急性影响。急性流出物毒性试验可用于评估接收河流、湖泊或河口中指定用途或水生生物损害的可能性。急性毒性试验不能提供关于是否会发生延迟效应的信息，尽管暴露后观察期（必要时适当喂食）可能会提供此类信息。 5.2 急性流出物试验结果可用于预测在可比条件下暴露可能对现场水生生物造成的急性影响，但( 1. )如果可能的话，活动的生物体可以避免暴露( 2. )对底栖生物的毒性可能取决于流出物成分在基质上的吸附或沉降，以及( 3. )流出物可能与接收水发生物理或化学相互作用。 5.3 急性流出物试验的结果可用于比较不同物种的急性敏感性和不同流出物的急性毒性，并研究各种环境因素对此类试验结果的影响。 5.4 急性试验通常是评估废水对水生生物影响的第一步。 5.5 急性流出物试验的结果将取决于温度、稀释水的组成、试验生物的条件、暴露技术和其他因素。

1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of an aqueous effluent on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, usually during 2 day to 4 day exposures, depending on the species, using the static, renewal, and flow-through techniques. These procedures will probably be useful for conducting acute toxicity tests on aqueous effluents with many other aquatic species, 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 acute toxicity tests on aqueous effluents. 1.3 This guide is based in large part on Guide E729 where addition details are provided for test elements that may be applicable to the ambient and effluent toxicity testing described in this method. The major differences between the two guides are ( 1 ) the maximum test concentration is 100 % effluent or ambient sample, ( 2 ) testing is not chemical-specific, and ( 3 ) the holding time of effluent and ambient samples is often considerably less than that for chemicals and other test materials. Because the sample is often a complex mixture of chemicals, analytical tests cannot generally be used to confirm exposure concentrations. 1.4 Selection of the technique to be used in a specific situation will depend upon the needs of the investigator and upon available resources. Static tests provide the most easily obtained measure of acute toxicity but should not last longer than 48 h. Renewal and flow-through tests may last longer than 48 h because the pH and concentrations of dissolved oxygen and effluent are maintained at desired levels and degradation and metabolic products are removed. Static tests might not be applicable to effluents that have a high oxygen demand or contain materials that ( 1 ) are highly volatile, ( 2 ) are rapidly biologically or chemically transformed in aqueous solutions, or ( 3 ) are removed from test solutions in substantial quantities by the test chambers or organisms during the test. Flow-through tests are generally preferable to renewal tests, although in some situations a renewal test might be more cost-effective than a flow-through test. 1.5 In the development of these procedures, an attempt was made to balance scientific and practical considerations and to ensure that the results will be sufficiently accurate and precise for the applications for which they are commonly used. A major consideration was that the common uses of the results of acute tests on effluents do not require or justify stricter requirements than those set forth in this guide. Although the tests may be improved by using more organisms, longer acclimation times, and so forth, the requirements presented in this guide should usually be sufficient. 1.6 Results of acute toxicity tests should usually be reported in terms of a median lethal concentration (LC50) or median effective concentration (EC50). In some situations, it might be necessary only to determine whether a specific concentration is acutely toxic to the test species or whether the LC50 or EC50 is above or below a specific concentration. 1.7 This guide is arranged as follows: Section Referenced Documents 2 Terminology 3 Summary of Guide 4 Significance and Use 5 Hazards 7 Apparatus 6 Facilities 6.1 Special Requirements 6.2 Construction Materials 6.3 Metering System 6.4 Test Chambers 6.5 Cleaning 6.6 Acceptability 6.7 Dilution Water 8 Requirements 8.1 Source 8.2 Treatment 8.3 Characterization 8.4 Effluent 9 Sampling Point 9.1 Collection 9.2 Preservation 9.3 Treatment 9.4 Test Concentration(s) 9.5 Test Organisms 10 Species 10.1 Age 10.2 Source 10.3 Care and Handling 10.4 Feeding 10.5 Disease Treatment 10.6 Holding 10.7 Acclimation 10.8 Quality 10.9 Procedure 11 Experimental Design 11.1 Dissolved Oxygen 11.2 Temperature 11.3 Loading 11.4 Beginning the Test 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 or Results 14 Report 15 1.8 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 7 . 1.9 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 An acute effluent toxicity test is conducted to obtain information concerning the immediate effects on test organisms of a short-term exposure to an effluent under specific experimental conditions. One can directly examine acute effects of complex mixtures of chemicals as occurs in effluents and some ambient waters. Acute effluent toxicity tests can be used to evaluate the potential for designated-use or aquatic life impairment in the receiving stream, lake, or estuary. An acute toxicity test does not provide information about whether delayed effects will occur, although a post-exposure observation period, with appropriate feeding if necessary, might provide such information. 5.2 Results of acute effluent tests might be used to predict acute effects likely to occur on aquatic organisms in field situations as a result of exposure under comparable conditions, except that ( 1 ) motile organisms might avoid exposure when possible, ( 2 ) toxicity to benthic species might be dependent on sorption or settling of components of the effluent onto the substrate, and ( 3 ) the effluent might physically or chemically interact with the receiving water. 5.3 Results of acute effluent tests might be used to compare the acute sensitivities of different species and the acute toxicities of different effluents, and to study the effects of various environmental factors on results of such tests. 5.4 Acute tests are usually the first step in evaluating the effects of an effluent on aquatic organisms. 5.5 Results of acute effluent tests will depend on the temperature, composition of the dilution water, condition of the test organisms, exposure technique, and other factors.