ASTM E1676-12(2021)
(Guide)Standard Guide for Conducting Laboratory Soil Toxicity or Bioaccumulation Tests with the Lumbricid Earthworm Eisenia Fetida and the Enchytraeid Potworm Enchytraeus albidus
Standard Guide for Conducting Laboratory Soil Toxicity or Bioaccumulation Tests with the Lumbricid Earthworm <emph type="ital">Eisenia Fetida</emph > and the Enchytraeid Potworm <emph type="ital">Enchytraeus albidus</emph >
SIGNIFICANCE AND USE
5.1 Soil toxicity tests provide information concerning the toxicity and bioavailability of chemicals associated with soils to terrestrial organisms. As important members of the soil fauna, lumbricid earthworms and enchytraeid potworms have a number of characteristics that make them appropriate organisms for use in the assessment of potentially hazardous soils. Earthworms may ingest large quantities of soil, have a close relationship with other soil biomasses (for example, invertebrates, roots, humus, litter, and microorganisms), constitute up to 92 % of the invertebrate biomass of soil, and are important in recycling nutrients (1, 2).4 Enchytraeids contribute up to 5.2 % of soil respiration, constitute the second-highest biomass in many soils (the highest in acid soils in which earthworms are lacking) and effect considerably nutrient cycling and community metabolism (3-5). Earthworms and potworms accumulate and are affected by a variety of organic and inorganic compounds (2-10, 11-14). In addition, earthworms and potworms are important in terrestrial food webs, constituting a food source for a very wide variety of organisms, including birds, mammals, reptiles, amphibians, fish, insects, nematodes, and centipedes (15, 16, 3). A major change in the abundance of soil invertebrates such as lumbricids or enchytraeids, either as a food source or as organisms functioning properly in trophic energy transfer and nutrient cycling, could have serious adverse ecological effects on the entire terrestrial system.
5.2 A number of species of lumbricids and enchytraeid worms have been used in field and laboratory investigations in the United States and Europe. Although the sensitivity of various lumbricid species to specific chemicals may vary, from their study of four species of earthworms (including E. fetida) exposed to ten organic compounds representing six classes of chemicals, Neuhauser, et al (7) suggest that the selection of earthworm test species does not affect the a...
SCOPE
1.1 This guide covers procedures for 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) from soil toxicity or bioaccumulation tests. The methods are designed to assess lethal or sublethal toxic effects on earthworms or bioaccumulation of contaminants in short-term tests (7 to 28 days) or on potworms in short to long-term tests (14 to 42 days) in terrestrial systems. Soils to be tested may be (1) reference soils or potentially toxic site soils; (2) artificial, reference, or site soils spiked with compounds; (3) site soils diluted with reference soils; or (4) site or reference soils diluted with artificial soil. Test procedures 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 also be useful for conducting soil toxicity tests with other lumbricid and enchytraeid 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 and bioaccumulation 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 c...
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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.
Designation: E1676 − 12 (Reapproved 2021)
Standard Guide for
Conducting Laboratory Soil Toxicity or Bioaccumulation
Tests with the Lumbricid Earthworm Eisenia Fetida and the
1
Enchytraeid Potworm Enchytraeus albidus
This standard is issued under the fixed designation E1676; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope soil characteristics (for example, particle size, organic matter
content, and clay content) are of interest or when there is a
1.1 This guide covers procedures for obtaining laboratory
need to test such materials as sewage sludge and oils. These
data to evaluate the adverse effects of contaminants (for
methods might also be useful for conducting bioaccumulation
example, chemicals or biomolecules) associated with soil to
tests.
earthworms (Family Lumbricidae) and potworms (Family
Enchytraeidae)fromsoiltoxicityorbioaccumulationtests.The
1.4 The results of toxicity tests with (1) materials (for
methods are designed to assess lethal or sublethal toxic effects
example, chemicals or waste mixtures) added experimentally
on earthworms or bioaccumulation of contaminants in short-
to artificial soil, reference soils, or site soils, (2) site soils
term tests (7 to 28 days) or on potworms in short to long-term
diluted with reference soils, and (3) site or reference soils
tests (14 to 42 days) in terrestrial systems. Soils to be tested
diluted with artificial soil, so as to create a series of
may be (1) reference soils or potentially toxic site soils; (2)
concentrations, may be reported in terms of an LC50 (median
artificial, reference, or site soils spiked with compounds; (3)
lethal concentration) and sometimes an EC50 (median effect
site soils diluted with reference soils; or (4) site or reference
concentration).Test results may be reported in terms of NOEC
soils diluted with artificial soil. Test procedures are described
(no observed effect concentration), LOEC (lowest observed
for the species Eisenia fetida (see Annex A1) and for the
effect concentration) or as an ECx (concentration where x%
species Enchytraeus albidus (see Annex A4). Methods de-
reduction of a biological effect occurs. Bioaccumulation test
scribed in this guide may also be useful for conducting soil
results are reported as the magnitude of contaminant concen-
toxicity tests with other lumbricid and enchytraeid terrestrial
tration above either the Day 0 tissue baseline analysis or the
species, although modifications may be necessary.
Day 28 tissues from the negative control or reference soil (that
is, 2x, 5x, 10x) (see A3.9).
1.2 Modification of these procedures might be justified by
special needs. The results of tests conducted using atypical
1.5 This guide is arranged as follows:
procedures may not be comparable to results using this guide.
Scope 1
Comparisonofresultsobtainedusingmodifiedandunmodified
Referenced Documents 2
versions of these procedures might provide useful information Terminology 3
Summary of Guide 4
concerning new concepts and procedures for conducting soil
Significance and Use 5
toxicity and bioaccumulation tests with terrestrial worms.
Interferences 6
Apparatus 7
1.3 The results from field-collected soils used in toxicity
Safety Precautions 8
tests to determine a spatial or temporal distribution of soil
Soil 9
Test Organism 10
toxicity may be reported in terms of the biological effects on
Procedure 11
survival or sublethal endpoints (see Section 14). These proce-
Analytical Methodology 12
dures can be used with appropriate modifications to conduct
Acceptability of Test 13
Calculation of Results 14
soil toxicity tests when factors such as temperature, pH, and
Report 15
Annexes
Annex A1. Eisenia fetida
1
Annex A2. Artificial Soil Composition
ThisguideisunderthejurisdictionofASTMCommitteeE50onEnvironmental
Annex A3. Bioaccumulation Testing Utilizing Eisenia fetida
Assessment, Risk Management and CorrectiveAction and is the direct responsibil-
Annex A4. Enchytraeid Reporduction Test (ERT)
ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.
References
An ASTM guide is defined as a series of options or instructions that do not
recommend a specific course of action.
1.6 The values stated in SI units are to be regarded as
Current edition approved Jan. 1, 2021. Published February 2021. Originally
standard. No other units of measurement are included in this
approved in 1995. Last previous edition approved in 2012 as E1676–12. DOI:
10.1520/E1676-1
...
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