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1.1 This guide covers procedures for obtaining laboratory data to evaluate the adverse effects of contaminants associated with soil on earthworms (Family Lumbricidae) from soil toxicity tests. The methods are designed to assess lethal or sublethal toxic effects on earthworms in short-term tests (7 to 28 days) in terrestrial systems. Soils to be tested may be ( ) reference soils or potentially toxic site soils; ( ) artificial, reference, or site soils spiked with compounds; ( ) site soils diluted with reference soils; or ( ) site or reference soils diluted with artificial soil. Test procedures are described for the species Eisenia foetida (see Annex A1). Methods described in this guide may also be useful for conducting soil toxicity tests with other terrestrial species, although modifications may be necessary.

1.2 Modification of these procedures might be justified by special needs. The results of tests conducted using atypical procedures may not be comparable to results using this guide. Comparison of results obtained using modified and unmodified versions of these procedures might provide useful information concerning new concepts and procedures for conducting soil toxicity tests with terrestrial worms.

1.3 The results from field-collected soils used in toxicity tests to determine a spatial or temporal distribution of soil toxicity may be reported in terms of the biological effects on survival or sublethal endpoints (see Section 14). These procedures can be used with appropriate modifications to conduct soil toxicity tests when factors such as temperature, pH, and soil characteristics (for example, particle size, organic matter content, and clay content) are of interest or when there is a need to test such materials as sewage sludge and oils. These methods might also be useful for conducting bioaccumulation tests.

1.4 The results of toxicity tests with ( ) materials (for example, chemicals or waste mixtures) added experimentally to artificial soil, reference soils, or site soils, ( ) site soils diluted with reference soils, and ( ) site or reference soils diluted with artificial soil, so as to create a series of concentrations, may be reported in terms of an LC50 (median lethal concentration) and sometimes an EC50 (median effect concentration). Test results may be reported in terms of NOEC (no observed effect concentration) and LOEC (lowest observed effect concentration).

1.5 This guide is arranged as follows:

Scope 1 Referenced Documents 2 Terminology 3 Summary of Guide 4 Significance and Use 5 Interferences 6 Apparatus 7 Safety Precautions 8 Soil 9 Test Organism 10 Procedure 11 Analytical Methodology 12 Acceptability of Test 13 Calculation of Results 14 Report 15 Annexes A1. Eisenia foetida A2. Artificial Soil Composition References

1.6 The values stated in SI units are to be regarded as the standard.

1.7 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. While some safety considerations are included in this guide, it is beyond the scope of this standard to encompass all safety requirements necessary to conduct soil toxicity tests. Specific precautionary statements are given in Section 8.