1.1This 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) although modifications might be necessary.
1.2Other 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.
1.3These 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.
1.4Results 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.
1.5This 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.
1.7This 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.