1.1 本指南描述了获取有关试验材料对四种咸水双壳类软体动物（太平洋牡蛎、， 巨牡蛎 桑伯格；东方牡蛎， 美洲牡蛎 Gmelin；quahog或硬蛤蜊， 硬壳蛤 林奈；和贻贝物种复合体( 贻贝属。 )包括蓝贻贝， 紫贻贝 林奈；地中海贻贝， 黄贻贝 拉马克； 北部湾贻贝， 托氏贻贝 在静态48小时暴露期间。这些程序可能有助于从其他双壳类动物的胚胎开始进行静态短期慢性毒性试验( 1. ) 2. 尽管可能需要修改。 1.2 根据特殊需要或情况，可能需要对这些程序进行其他修改。虽然使用适合特定物种或特殊需要和情况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，可能会提供有关从双壳类软体动物胚胎开始进行48小时急性试验的新概念和程序的有用信息。 1.3 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。经过适当修改，这些程序可用于对温度、溶解氧和pH值以及含水废水等材料进行急性试验（另请参阅指南） E1192 )、沥滤液、油、颗粒物、沉积物和地表水。对于具有高需氧量、高挥发性、在水溶液中快速生物或化学转化、或在试验过程中被试验室或生物体从试验溶液中大量去除的材料，更新试验可能优于静态试验。 1.4 双壳类软体动物胚胎毒性试验结果通常应根据未完全发育和死亡的生物体总数报告为EC50。 也可以只根据死亡报告LC50。在某些情况下，可能只需要确定特定浓度是否对胚胎有毒，或者EC50是否高于或低于特定浓度。 1.5 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 6. 仪器 7. 设施 7.1 建筑材料 7.2 试验室 7.3 打扫 7.4 可接受性 7.5 稀释水 8. 要求 8.1 来源 8.2 治疗 8.3 刻画 8.4 试验材料 9 全体的 9.1 储备溶液 9.2 试验浓度 9.3 测试生物体 10 种 10.1 年龄 10.2 处理 10.3 种苗来源和条件 10.4 产卵和受精 10.5 质量 10.6 程序 11 实验设计 11.1 溶解氧 11.2 温度 11.3 开始测试 11.4 喂养 11.5 试验持续时间 11.6 生物学数据 11.7 其他测量 11.8 分析方法 12 测试的可接受性 13 计算结果 14 汇报 15 附件 附件A1 1.6 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和健康实践，并确定监管限制的适用性。 第节给出了具体的危险说明 6. . 1.7 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 进行急性毒性试验，以评估生物体在特定实验条件下短期暴露于试验材料的影响。急性毒性试验不提供是否会发生延迟效应的信息，通常评估对生存的影响。慢性试验通常持续时间较长，包括亚致死终点，以评估暴露期后可能对人群产生的影响。 由于双壳类胚胎发育试验包括亚致死终点，但持续时间短，因此这些试验被视为短期慢性试验。 5.2 由于胚胎和幼虫通常被认为是这些双壳类软体动物物种最敏感的生命阶段，并且由于这些物种在商业和娱乐上都很重要，因此这些急性试验的结果通常被认为是海水软体动物物种对污染物浓度可接受性的良好指示。 在评估材料对其他盐水生物的危害时，通常认为这些急性毒性试验的结果是一个重要的考虑因素（见指南） E1023 )或者在推导咸水生物的水质标准时 ( 3. ) . 5.3 短期慢性毒性试验的结果可用于预测在类似条件下暴露可能对现场水生生物产生的影响，但对底栖物种的毒性可能取决于试验材料在基质上的吸附或沉降。 5.4 短期慢性试验的结果可用于比较不同物种对不同试验材料的敏感性，并确定各种环境因素对此类试验结果的影响。 5.5 短期慢性毒性试验的结果可能有助于研究试验材料的生物可用性和结构-活性关系。 5.6 任何毒性试验的结果将取决于温度、稀释水的成分、试验生物体的状况和其他因素。 5.7 短期慢性毒性试验的结果可用于预测暴露于通过水柱处理的疏浚沉积物悬浮颗粒物的水生生物可能产生的影响。 5.8 短期慢性毒性试验的结果可用于预测暴露在层状完整沉积物中的水生生物可能受到的影响。

1.1 This guide describes procedures for obtaining laboratory data concerning the acute effects of a test material on embryos and the resulting larvae of four species of saltwater bivalve molluscs (Pacific oyster, Crassostrea gigas Thunberg; eastern oyster, Crassostrea virginica Gmelin; quahog or hard clam, Mercenaria mercenaria Linnaeus; and the mussel species complex ( Mytilus spp. ) including the blue mussel, Mytilus edulis Linnaeus; the Mediterranean mussel, Mytilus galloprovincialis Lamark; and the Northern Bay Mussel, Mytilus trossulus Gould) during static 48-h exposures. These procedures will probably be useful for conducting static short-term chronic toxicity tests starting with embryos of other bivalve species ( 1 ) 2 although modifications might be necessary. 1.2 Other modifications of these procedures might be justified by special needs or circumstances. Although using procedures appropriate to a particular species or special needs and circumstances is more important than following prescribed procedures, results of tests conducted by using unusual procedures are not likely to be comparable to results of many other tests. Comparison of results obtained by using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting 48-h acute tests starting with embryos of bivalve molluscs. 1.3 These procedures are applicable to most chemicals, either individually or in formulations, commercial products, or known mixtures. With appropriate modifications these procedures can be used to conduct acute tests on temperature, dissolved oxygen, and pH and on such materials as aqueous effluents (see also Guide E1192 ), leachates, oils, particulate matter, sediments, and surface waters. Renewal tests might be preferable to static tests for materials that have a high oxygen demand, are highly volatile, are rapidly biologically or chemically transformed in aqueous solution, or are removed from test solutions in substantial quantities by the test chambers or organisms during the test. 1.4 Results of toxicity tests with embryos of bivalve molluscs should usually be reported as the EC50 based on the total incompletely developed and dead organisms. It might also be desirable to report the LC50 based only on death. In some situations, it might only be necessary to determine whether a specific concentration is toxic to embryos or whether the EC50 is above or below a specific concentration. 1.5 This guide is arranged as follows: Section Referenced Documents 2 Terminology 3 Summary of Guide 4 Significance and Use 5 Hazards 6 Apparatus 7 Facilities 7.1 Construction Materials 7.2 Test Chambers 7.3 Cleaning 7.4 Acceptability 7.5 Dilution Water 8 Requirements 8.1 Source 8.2 Treatments 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 Handling 10.3 Brood Stock Source and Condition 10.4 Spawning and Fertilization 10.5 Quality 10.6 Procedure 11 Experimental Design 11.1 Dissolved Oxygen 11.2 Temperature 11.3 Beginning the Test 11.4 Feeding 11.5 Duration of Test 11.6 Biological Data 11.7 Other Measurements 11.8 Analytical Methods 12 Acceptability of Test 13 Calculation of Results 14 Report 15 Annex Annex A1 1.6 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 and health practices and determine the applicability of regulatory limitations prior to use. Specific hazard statements are given in Section 6 . 1.7 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 toxicity test is conducted to assess the effects of a short term exposure of organisms to a test material under specific experimental conditions. An acute toxicity test does not provide information concerning whether delayed effects will occur and typically evaluates effects on survival. A chronic test is typically longer in duration and includes a sublethal endpoint to assess effects on a population that might occur beyond the exposure period. Because the bivalve embryo development test includes a sublethal endpoint, but is also short in duration, these tests are considered to be short-term chronic tests. 5.2 Because embryos and larvae are usually assumed to be the most sensitive life stages of these bivalve mollusc species and because these species are commercially and recreationally important, results of these acute tests are often considered to be a good indication of the acceptability of pollutant concentrations to saltwater molluscan species in general. Results of these acute toxicity tests are often assumed to be an important consideration when assessing the hazard of materials to other saltwater organisms (see Guide E1023 ) or when deriving water quality criteria for saltwater organisms ( 3 ) . 5.3 Results of short-term chronic toxicity tests might be used to predict effects likely to occur to aquatic organisms in field situations as a result of exposure under comparable conditions, except that toxicity to benthic species might depend on sorption or settling of the test material onto the substrate. 5.4 Results of short-term chronic tests might be used to compare the sensitivities of different species to different test materials, and to determine the effects of various environmental factors on results of such tests. 5.5 Results of short-term chronic toxicity tests might be useful for studying biological availability of, and structure activity relationships between, test materials. 5.6 Results of any toxicity test will depend on temperature, composition of the dilution water, condition of the test organisms, and other factors. 5.7 Results of short-term chronic toxicity tests might be used to predict effects likely to occur to aquatic organisms exposed to suspended particulates of dredged sediments disposed through the water column. 5.8 Results of short-term chronic toxicity tests might be used to predict effects likely to occur to aquatic organisms exposed to a bedded whole sediments.