1、Designation: E1676 04 E1676 12Standard Guide forConducting Laboratory Soil Toxicity or BioaccumulationTests with the Lumbricid Earthworm Eisenia Fetida and theEnchytraeid Potworm Enchytraeus albidus1This standard is issued under the fixed designation E1676; the number immediately following the desig
2、nation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This guide covers procedures fo
3、r obtaining laboratory data to evaluate the adverse effects of contaminants (for example,chemicals or biomolecules) associated with soil to earthworms (Family Lumbricidae) and potworms (Family Enchytraeidae) fromsoil toxicity or bioaccumulation tests. The methods are designed to assess lethal or sub
4、lethal toxic effects on earthworms orbioaccumulation of contaminants in short-term tests (7 to 28 days) or on potworms in short to long-term tests (14 to 42 days) interrestrial systems. Soils to be tested may be (1) reference soils or potentially toxic site soils; (2) artificial, reference, or site
5、soilsspiked with compounds; (3) site soils diluted with reference soils; or (4) site or reference soils diluted with artificial soil. Testprocedures are described for the species Eisenia fetida (see Annex A1) and for the species Enchytraeus albidus (see Annex A4).Methods described in this guide may
6、also be useful for conducting soil toxicity tests with other lumbricid and enchytraeidterrestrial species, although modifications may be necessary.1.2 Modification of these procedures might be justified by special needs. The results of tests conducted using atypicalprocedures may not be comparable t
7、o results using this guide. Comparison of results obtained using modified and unmodifiedversions of these procedures might provide useful information concerning new concepts and procedures for conducting soiltoxicity and bioaccumulation tests with terrestrial worms.1.3 The results from field-collect
8、ed soils used in toxicity tests to determine a spatial or temporal distribution of soil toxicity maybe reported in terms of the biological effects on survival or sublethal endpoints (see Section 14). These procedures can be usedwith appropriate modifications to conduct soil toxicity tests when facto
9、rs such as temperature, pH, and soil characteristics (forexample, particle size, organic matter content, and clay content) are of interest or when there is a need to test such materials assewage sludge and oils. These methods might also be useful for conducting bioaccumulation tests.1.4 The results
10、of toxicity tests with (1) materials (for example, chemicals or waste mixtures) added experimentally to artificialsoil, reference soils, or site soils, (2) site soils diluted with reference soils, and (3) site or reference soils diluted with artificial soil,so as to create a series of concentrations
11、, may be reported in terms of an LC50 (median lethal concentration) and sometimes anEC50 (median effect concentration). Test results may be reported in terms of NOEC (no observed effect concentration), LOEC(lowest observed effect concentration) or as an ECx (concentration where x % reduction of a bi
12、ological effect occurs.Bioaccumulation test results are reported as the magnitude of contaminant concentration above either the Day 0 tissue baselineanalysis or the Day 28 tissues from the negative control or reference soil (that is, 2, 5, 10)2x, 5x, 10x) (see A3.9).1.5 This guide is arranged as fol
13、lows:Scope 1Referenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Interferences 6Apparatus 7Safety Precautions 8Soil 9Test Organism 101 This guide is under the jurisdiction of ASTM Committee E47 on Biological Effects and Environmental Fateand is the direct responsibility of Subc
14、ommittee E47.02 onTerrestrial Assessment and Toxicology.An ASTM guide is defined as a series of options or instructions that do not recommend a specific course of action.Current edition approved April 1, 2004Nov. 1, 2012. Published May 2004December 2012. Originally approved in 1995. Last previous ed
15、ition approved in 19972004 asE1676 97.E167604. DOI: 10.1520/E1676-04.10.1520/E1676-12.This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Becauseit may not be technically possible to
16、adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current versionof the standard as published by ASTM is to be considered the official document.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Consh
17、ohocken, PA 19428-2959. United States1Procedure 11Analytical Methodology 12Acceptability of Test 13Calculation of Results 14Report 15AnnexesAnnex A1. Eisenia fetidaAnnex A2. Artificial Soil CompositionAnnex A3. Bioaccumulation Testing Utilizing Eisenia fetidaAnnex A4. Enchytraeid Reporduction Test (
18、ERT)References1.6 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 responsibilityof the user of this standard to establish appropriate safety and health practices and
19、determine the applicability of regulatorylimitations prior to use. While some safety considerations are included in this guide, it is beyond the scope of this standard toencompass all safety requirements necessary to conduct soil toxicity tests.Specific precautionary statements are given in Section8
20、.2. Referenced Documents2.1 ASTM Standards:2D653 Terminology Relating to Soil, Rock, and Contained FluidsD4447 Guide for Disposal of Laboratory Chemicals and SamplesE380 Practice for Use of the International System of Units (SI) (the Modernized Metric System) (Withdrawn 1997)3E943 Terminology Relati
21、ng to Biological Effects and Environmental FateE1023 Guide for Assessing the Hazard of a Material to Aquatic Organisms and Their UsesE1383 Guide for Conducting Sediment Toxicity Tests with Freshwater Invertebrates (Withdrawn 1995)3E1688 Guide for Determination of the Bioaccumulation of Sediment-Asso
22、ciated Contaminants by Benthic InvertebratesE1706 Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Freshwater Invertebrates3. Terminology3.1 Definitions:3.1.1 The words “must,” “should,” “may,”“ can,” and “might” have very specific meanings in this guide. “Must” is use
23、d toexpress an absolute requirement, that is, to state that the test must be designed to satisfy the specified condition, unless the purposeof the test requires a different design. “Must” is used only in connection with the factors that relate directly to the acceptabilityof the test (see Section 13
24、). “Should” is used to state that the specified condition is recommended and ought to be met if possible.Although a violation of one “should” is rarely a serious matter, the violation of several will often render the results questionable.Terms such as “is desirable,” “is often desirable,” and “might
25、 be desirable” are used in connection with less important factors.“May” is used to mean “is (are) allowed to,” “can” is used to mean “is (are) able to,” and “might” is used to mean “could possibly.”Thus, the classic distinction between “may” and “can” is preserved, and “might” is never used as a syn
26、onym for either “may” or“can.”3.1.2 For definitions of terms used in this guide, refer to Terminology E943 and Guide E1023. For an explanation of units andsymbols, refer to Practice E380.3.2 Definitions of Terms Specific to This Standard:3.2.1 artificial soila synthetic soil, prepared with a specifi
27、c formulation, designed to simulate a natural soil (see Annex A2).Artificial soil may be used as a diluent medium to prepare concentrations of site or reference soil and may be used as a negativecontrol medium.3.2.2 batchthe total amount of test soil prepared for each concentration in a test. A batc
28、h is any hydrated test soil ready forseparation into replicates.3.2.3 bioaccumulationthe net accumulation of a substance by an organism as a result of uptake from all environmentalsources. (See Guide E1688.)3.2.4 bioaccumulation factor (BAF)the ratio of tissue residue to sediment or soil contaminant
29、 concentration at steady-state.(See Guide E1688.)3.2.5 bioaccumulation potentiala qualitative assessment of whether a contaminant in a particular sediment or soil isbioavailable. (See Guide E1688.)3.2.6 bioconcentrationthe net assimilation of a substance by an organism as a result of uptake directly
30、 from aqueous solution.(See Guide E1688.)2 For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at serviceastm.org. For Annual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.3 The last approv
31、ed version of this historical standard is referenced on www.astm.org.E1676 1223.2.7 bioconcentration factor (BCF)the ratio of tissue residue to water contaminant concentration as steady-state. (See GuideE1688.)3.2.8 biota-sediment accumulation factor (BSAF) the ratio of lipid-normalized tissue resid
32、ue to organic carbon-normalizedsediment contaminant concentration at steady state, with units of g-carbon/g-lipid. (See Guide E1688.)3.2.9 clitellumthe fleshy “ring” or “saddle” of glandular tissue found on certain mid-body segments of oligochaete(Lumbricidae and Enchytraeidae) worms. It is the most
33、 visible feature of an adult earthworm or potworm and secretes the cocooninto which eggs and sperm are deposited.3.2.10 concentrationthe ratio of the weight of test materials to the weight of soil (artificial, reference, or site), usuallyexpressed on a dry weight basis as percent or milligram/kilogr
34、am.3.2.11 depurationloss of a substance from an organism as a result of any active (for example, metabolic breakdown) orpassive process.3.2.12 diluent soilthe artificial or reference soil used to dilute site soils.3.2.13 enchytraeidpotworm members of the Family Enchytraeidae of the Class Oligochaeta
35、 of the Phylum Annelida.3.2.14 hydration waterwater used to hydrate test soils to create an environment with a moisture level suitable for the speciesbeing tested. The water used for hydration is often test water (see 3.2.27); however, depending on the nature of the test beingimplemented, site surfa
36、ce water or groundwater may also be utilized for hydration.3.2.15 lumbricidearthworm members of the Family Lubricidae of the Class Oligochaeta of the Phylum Annelida.3.2.16 negative control soilartificial or reference soil to be used for evaluating the acceptability of a test.3.2.17 reference soila
37、field-collected soil that has physicochemical and biological properties as similar as possible to the sitesoil but does not contain the potentially toxic compounds of the site soil. It is used to describe matrix effects on the test in question.It may be used as a diluent medium to prepare concentrat
38、ions of site soil and may be used as a negative control medium.3.2.18 sampling stationa specific location, within a site or sampling unit, depending on the field study design, at which soilis collected for chemical, physical, and biological evaluation.3.2.19 sampling unitan area of land within a sit
39、e distinguished by habitat and topography.3.2.20 sitea delineated tract of land that is being considered as a study area, usually from the standpoint of its being potentiallyaffected by xenobiotics.3.2.21 site soila soil collected from the field to be evaluated for potential toxicity. A site soil ma
40、y be a naturally occurring soilor one that has been influenced by xenobiotics.3.2.22 soilsediments or other unconsolidated accumulations of solid particles produced by the physical and chemicaldisintegration of rocks, and that may or may not contain organic material. (See Terminology D653.)3.2.23 sp
41、ikingthe experimental addition of a test material to an artificial, site, or reference soil, such that the toxicity of thematerial added can be determined. After the test material is added, which may involve a solvent carrier, the soil is mixed thoroughlyto distribute the test material evenly throug
42、hout the soil.3.2.24 test chamberan enclosed space or compartment in which environmental parameters such as temperature and lightingare controlled (for example, incubator or modified room). Test containers are placed in the test chamber for biological evaluation.3.2.25 test containerthe experimental
43、 unit; the smallest physical entity to which treatments can be assigned independently.3.2.26 test soila soil prepared to receive a test organism. Site or reference soil mixed with artificial soil or reference soil mixedwith site soil in known concentrations for evaluation are test soils. Artificial,
44、 site, or reference soils spiked with test materials suchas chemicals, oils, or manufacturing products are test soils. Once a site, reference, or artificial soil is hydrated, even though it isnot mixed with artificial or reference soil or spiked with a material, it may be called a test soil.3.2.27 t
45、est waterwater used to prepare stock solutions, rinse test organisms, rinse glassware, and apparatus or for any otherpurpose associated with the test procedures or culture of the test organism. Test water must be deionized or distilled water or better,such as reagent-grade water produced by a system
46、 of reverse osmosis, carbon, and ion-exchange cartridges.4. Summary of Guide4.1 The toxicity of test soils or the bioavailability of contaminants are assessed during the continuous exposure of terrestrialorganisms. Soils tested may be the following: (1) soils collected from potentially contaminated
47、sites, (2 ) soils collected fromreference sites, (3) artificial soil (see Annex A2) spiked with compounds, (4) site soil spiked with compounds, (5) reference soilspiked with compounds, (6) site soil diluted with artificial soil, (7) site soil diluted with reference soil, or (8) reference soil dilute
48、dwith artificial soil. A negative control of artificial or reference soil is used for the following: (1) to yield a measure of theacceptability of the test; (2) to provide evidence of the health and relative quality of the test organisms; (3) to determine thesuitability of test conditions, food, and
49、 handling procedures; and (4) to provide a basis for interpreting data obtained from the testE1676 123soils. Specified data are obtained to determine the toxic effects on survival or sublethal endpoints for 7 to 28-day exposures orcontainment bioaccumulation for 28-day exposures to terrestrial lumbricids and the toxic effects on survival or sublethal endpointsfor 4 to 42-day exposures to enchytraeids.4.2 Summary of ChangesThis current version of the standard is a revision of the E1676-97 version. Changes made since 1997involve toxicity testing pro
