ASTM E1367-03(2023)

Standard Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Estuarine and Marine Invertebrates

SIGNIFICANCE AND USE
5.1 General:  
5.1.1 Sediment provides habitat for many aquatic organisms and is a major repository for many of the more persistent chemicals that are introduced into surface waters. In the aquatic environment, most anthropogenic chemicals and waste materials including toxic organic and inorganic chemicals eventually accumulate in sediment. Mounting evidences exists of environmental degradation in areas where USEPA Water Quality Criteria (WQC; Stephan et al.(66)) are not exceeded, yet organisms in or near sediments are adversely affected Chapman, 1989  (67). The WQC were developed to protect organisms in the water column and were not directed toward protecting organisms in sediment. Concentrations of contaminants in sediment may be several orders of magnitude higher than in the overlying water; however, whole sediment concentrations have not been strongly correlated to bioavailability Burton, 1991 (68). Partitioning or sorption of a compound between water and sediment may depend on many factors including: aqueous solubility, pH, redox, affinity for sediment organic carbon and dissolved organic carbon, grain size of the sediment, sediment mineral constituents (oxides of iron, manganese, and aluminum), and the quantity of acid volatile sulfides in sediment Di Toro et al. 1991(69) Giesy et al. 1988 (70). Although certain chemicals are highly sorbed to sediment, these compounds may still be available to the biota. Chemicals in sediments may be directly toxic to aquatic life or can be a source of chemicals for bioaccumulation in the food chain.  
5.1.2 The objective of a sediment test is to determine whether chemicals in sediment are harmful to or are bioaccumulated by benthic organisms. The tests can be used to measure interactive toxic effects of complex chemical mixtures in sediment. Furthermore, knowledge of specific pathways of interactions among sediments and test organisms is not necessary to conduct the tests Kemp et al. 1988, (71). Sediment tests can be used ...
SCOPE
1.1 This test method covers procedures for testing estuarine or marine organisms in the laboratory to evaluate the toxicity of contaminants associated with whole sediments. Sediments may be collected from the field or spiked with compounds in the laboratory. General guidance is presented in Sections 1 – 15 for conducting sediment toxicity tests with estuarine or marine amphipods. Specific guidance for conducting 10-d sediment toxicity tests with estuarine or marine amphipods is outlined in Annex A1 and specific guidance for conducting 28-d sediment toxicity tests with  Leptocheirus plumulosus is outlined in Annex A2.  
1.2 Procedures are described for testing estuarine or marine amphipod crustaceans in 10-d laboratory exposures to evaluate the toxicity of contaminants associated with whole sediments (Annex A1; USEPA 1994a (1)). Sediments may be collected from the field or spiked with compounds in the laboratory. A toxicity method is outlined for four species of estuarine or marine sediment-burrowing amphipods found within United States coastal waters. The species are Ampelisca abdita, a marine species that inhabits marine and mesohaline portions of the Atlantic coast, the Gulf of Mexico, and San Francisco Bay; Eohaustorius estuarius, a Pacific coast estuarine species; Leptocheirus plumulosus, an Atlantic coast estuarine species; and Rhepoxynius abronius , a Pacific coast marine species. Generally, the method described may be applied to all four species, although acclimation procedures and some test conditions (that is, temperature and salinity) will be species-specific (Sections 12 and Annex A1). The toxicity test is conducted in 1-L glass chambers containing 175 mL of sediment and 775 mL of overlying seawater. Exposure is static (that is, water is not renewed), and the animals are not fed over the 10-d exposure period. The endpoint in the toxicity test is survival with reburial of surviving amphipods as an additional m...

General Information

• Status: Published
• Publication Date: 31-Dec-2022
• ICS: 07.060 - Geology. Meteorology. Hydrology
• Technical Committee: E50 - Environmental Assessment, Risk Management and Corrective Action
• Drafting Committee: E50.47 - Biological Effects and Environmental Fate

Relations

• Refers: ASTM E1402-13(2023) - Standard Guide for Sampling Design
• Effective Date: 01-Nov-2023
• Refers: ASTM E141-10(2023) - Standard Practice for Acceptance of Evidence Based on the Results of Probability Sampling
• Effective Date: 01-Nov-2023
• Refers: ASTM E456-13a(2022)e1 - Standard Terminology Relating to Quality and Statistics
• Effective Date: 01-Apr-2022
• Refers: ASTM D1129-13(2020)e2 - Standard Terminology Relating to Water
• Effective Date: 01-May-2020
• Refers: ASTM E1688-19 - Standard Guide for Determination of the Bioaccumulation of Sediment-Associated Contaminants by Benthic Invertebrates
• Effective Date: 01-Dec-2019
• Refers: ASTM E1706-19 - Standard Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Freshwater Invertebrates
• Effective Date: 01-Apr-2019
• Refers: ASTM E1850-04(2019) - Standard Guide for Selection of Resident Species as Test Organisms for Aquatic and Sediment Toxicity Tests
• Effective Date: 01-Feb-2019
• Refers: ASTM E1402-13(2018) - Standard Guide for Sampling Design
• Effective Date: 01-Nov-2018
• Refers: ASTM E141-10(2018) - Standard Practice for Acceptance of Evidence Based on the Results of Probability Sampling
• Effective Date: 01-Apr-2018
• Refers: ASTM E456-13A(2017)e3 - Standard Terminology Relating to Quality and Statistics
• Effective Date: 01-Oct-2017
• Refers: ASTM E456-13A(2017)e1 - Standard Terminology Relating to Quality and Statistics
• Effective Date: 01-Oct-2017
• Refers: ASTM E178-16 - Standard Practice for Dealing With Outlying Observations
• Effective Date: 01-Jun-2016
• Refers: ASTM E1688-10(2016) - Standard Guide for Determination of the Bioaccumulation of Sediment-Associated Contaminants by Benthic Invertebrates
• Effective Date: 01-Feb-2016
• Refers: ASTM E1325-15 - Standard Terminology Relating to Design of Experiments
• Effective Date: 01-Oct-2015
• Refers: ASTM E177-14 - Standard Practice for Use of the Terms Precision and Bias in ASTM Test Methods
• Effective Date: 01-May-2014

Overview

ASTM E1367-03(2023), titled Standard Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Estuarine and Marine Invertebrates, establishes laboratory procedures for evaluating the toxicity of contaminants present in whole sediments. Developed by ASTM International, this standard is a key resource for environmental assessment, sediment quality monitoring, and ecological risk management in aquatic environments. By focusing on both acute and chronic effects on estuarine and marine invertebrates-particularly amphipods-this method provides critical data for environmental scientists, regulators, and industry professionals concerned with sediment contamination.

Sediments act as major habitats for benthic (bottom-dwelling) organisms and serve as repositories for persistent chemical pollutants. Contaminants in sediments can reach levels much higher than in the overlying water, posing risks not just to benthic organisms but also to the wider aquatic food web through direct toxicity and bioaccumulation. This standard provides a consistent, scientifically validated approach to assess these risks.

Key Topics

• Purpose and Significance

  • Assesses the potential harm of sediment-associated chemicals to estuarine and marine invertebrates.
  • Addresses gaps left by traditional Water Quality Criteria, which focus mainly on water-column protection.

• Test Organisms

  • Focuses on sediment-burrowing amphipod species commonly found along U.S. coastal waters:
    • Ampelisca abdita
    • Eohaustorius estuarius
    • Leptocheirus plumulosus
    • Rhepoxynius abronius
  • Endpoints include survival (for 10-day exposures) and survival, growth, and reproduction (for 28-day exposures).

• Test Procedures

  • Includes guidance for sediment collection (from field or laboratory-spiked), test setup, exposure conditions, and result interpretation.
  • Tests are performed in controlled laboratory conditions using glass chambers with specific sediment and seawater ratios.

• Factors Influencing Toxicity

  • Considers sediment characteristics: grain size, organic carbon content, pH, redox potential, and salinity.
  • Assesses both lethal (mortality) and sublethal (growth and reproduction) effects for comprehensive risk assessment.

• Quality Assurance

  • Emphasizes the necessity for standardized methods and quality control, including interlaboratory studies and performance criteria (minimum survival rates in controls).

Applications

• Contaminated Sediment Assessment

  • Used by environmental agencies, consulting firms, and researchers to evaluate the toxicity of sediments in marine and estuarine settings.
  • Supports identification of contaminated sites, "hot spots," and areas needing remediation.

• Regulatory Compliance

  • Informs sediment management and dredging activities by providing data for regulatory decision making.
  • Supports risk assessments required by environmental regulations at local, national, and international levels.

• Ecological Risk and Restoration

  • Contributes to habitat and benthic community health assessments.
  • Helps prioritize sediment remediation projects and monitor recovery following cleanup efforts.

• Research and Method Development

  • Offers a foundation for future method enhancements, including tests for different species or alternative exposure durations.

Related Standards

• ASTM E1706 – Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Freshwater Invertebrates
• ASTM E1391 – Guide for Collection, Storage, Characterization, and Manipulation of Sediments for Toxicological Testing
• ASTM E1525 – Guide for Designing Biological Tests with Sediments
• ASTM E1611 – Guide for Conducting Sediment Toxicity Tests with Polychaetous Annelids
• ASTM E1688 – Guide for Determination of the Bioaccumulation of Sediment-Associated Contaminants by Benthic Invertebrates

ASTM E1367 is widely recognized and aligned with international standardization principles, making it an essential reference for professionals involved in sediment toxicity testing and the protection of aquatic ecosystems.

Standards Content (Sample)

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: E1367 − 03 (Reapproved 2023)
Standard Test Method for
Measuring the Toxicity of Sediment-Associated
Contaminants with Estuarine and Marine Invertebrates
This standard is issued under the fixed designation E1367; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope* species (for R. abronius and E. estuarius). Performance criteria
established for this test include the average survival of amphi-
1.1 This test method covers procedures for testing estuarine
pods in negative control treatment must be greater than or
or marine organisms in the laboratory to evaluate the toxicity
equal to 90 %. Procedures are described for use with sediments
of contaminants associated with whole sediments. Sediments
o
with pore-water salinity ranging from >0 ⁄oo to fully marine.
may be collected from the field or spiked with compounds in
the laboratory. General guidance is presented in Sections 1 – 15
1.3 A procedure is also described for determining the
for conducting sediment toxicity tests with estuarine or marine
chronic toxicity of contaminants associated with whole sedi-
amphipods. Specific guidance for conducting 10-d sediment
ments with the amphipod Leptocheirus plumulosus in labora-
toxicity tests with estuarine or marine amphipods is outlined in
tory exposures (Annex A2; USEPA-USACE 2001(2)). The
Annex A1 and specific guidance for conducting 28-d sediment
toxicity test is conducted for 28 d in 1-L glass chambers
toxicity tests with Leptocheirus plumulosus is outlined in
containing 175 mL of sediment and about 775 mL of overlying
Annex A2.
water. Test temperature is 25° 6 2 °C, and the recommended
o o
1.2 Procedures are described for testing estuarine or marine
overlying water salinity is 5 ⁄oo 6 2 ⁄oo (for test sediment with
o o o o
amphipod crustaceans in 10-d laboratory exposures to evaluate
pore water at 1 ⁄oo to 10 ⁄oo) or 20 ⁄oo 6 2 ⁄oo (for test
o
the toxicity of contaminants associated with whole sediments
sediment with pore water >10 ⁄oo). Four hundred millilitres of
(Annex A1; USEPA 1994a (1)). Sediments may be collected
overlying water is renewed three times per week, at which
from the field or spiked with compounds in the laboratory. A
times test organisms are fed. The endpoints in the toxicity test
toxicity method is outlined for four species of estuarine or
are survival, growth, and reproduction of amphipods. Perfor-
marine sediment-burrowing amphipods found within United
mance criteria established for this test include the average
States coastal waters. The species are Ampelisca abdita, a
survival of amphipods in negative control treatment must be
marine species that inhabits marine and mesohaline portions of
greater than or equal to 80 % and there must be measurable
the Atlantic coast, the Gulf of Mexico, and San Francisco Bay;
growth and reproduction in all replicates of the negative
Eohaustorius estuarius, a Pacific coast estuarine species;
control treatment. This test is applicable for use with sediments
Leptocheirus plumulosus, an Atlantic coast estuarine species;
from oligohaline to fully marine environments, with a silt
and Rhepoxynius abronius, a Pacific coast marine species.
content greater than 5 % and a clay content less than 85 %.
Generally, the method described may be applied to all four
o
species, although acclimation procedures and some test condi- 1.4 A salinity of 5 or 20 ⁄oo is recommended for routine
tions (that is, temperature and salinity) will be species-specific application of 28-d test with L. plumulosus (Annex A2;
o
(Sections 12 and Annex A1). The toxicity test is conducted in USEPA-USACE 2001 (2)) and a salinity of 20 ⁄oo is recom-
1-L glass chambers containing 175 mL of sediment and 775 mended for routine application of the 10-d test with E.
mL of overlying seawater. Exposure is static (that is, water is
estuarius or L. plumulosus (Annex A1). However, the salinity
not renewed), and the animals are not fed over the 10-d of the overlying water for tests with these two species can be
exposure period. The endpoint in the toxicity test is survival
adjusted to a specific salinity of interest (for example, salinity
with reburial of surviving amphipods as an additional measure-
representative of site of interest or the objective of the study
ment that can be used as an endpoint for some of the test
may be to evaluate the influence of salinity on the bioavail-
ability of chemicals in sediment). More importantly, the
salinity tested must be within the tolerance range of the test
This test method is under the jurisdiction of ASTM Committee E50 on
organisms (as outlined in Annex A1 and Annex A2). If tests are
Environmental Assessment, Risk Management and Corrective Action and is the
direct responsibility of Subcommittee E50.47 on Biological Effects and Environ-
conducted with procedures different from those described in
mental Fate.
1.3 or in Table A1.1 (for example, different salinity, lighting,
Current edition approved Jan. 1, 2023. Published March 2023. Originally
temperature, feeding conditions), additional tests are required
approved in 1990. Last previous edition approved in 2014 as E1367 – 03 (2014).
DOI: 10.1520/E1367-03R23. to determine comparability of results (1.10). If there is not a
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1367 − 03 (2023)
TABLE 1 Rating of Selection Criteria for Estuarine or Marine Amphipod Sediment Toxicity Testing
A “+” or “−” Rating Indicates a Positive or Negative Attribute
Ampelisca Eohaustorius Leptocheirus Rhepoxynius
Criterion
abdita estuarius plumulosus abronius
Relative sensitivity toxicity data base + + + +
Round-robin studies conducted + + + +
Contact with sediment + + + +
Laboratory culture +/- - + -
Taxonomic identification + + + +
Ecological importance + + + +
Geographical distribution ATL, PAC, GOM PAC ATL PAC
Sediment physicochemical tolerance + + + +
A
Response confirmed with benthos populations + + + +
Peer reviewed + + + +
Endpoints monitored Survival Survival, reburial Survival Survival, reburial
A
Anderson et al. (2001 (14)).
ATL = Atlantic Coast, PAC = Pacific Coast, GOM= Gulf of Mexico
need to make comparisons among studies, then the test could also needed to link the toxicity test endpoints to a field-
be conducted just at a selected salinity for the sediment of validated population model of L. plumulosus that would then
interest. generate estimates of population-level responses of the amphi-
pod to test sediments and thereby provide additional ecologi-
1.5 Future revisions of this standard may include additional
cally relevant interpretive guidance for the laboratory toxicity
annexes describing whole-sediment toxicity tests with other
test.
groups of estuarine or marine invertebrates (for example,
information presented in Guide E1611 on sediment testing with
1.9 This standard outlines specific test methods for evalu-
polychaetes could be added as an annex to future revisions to
ating the toxicity of sediments with A. abdita, E. estuarius, L.
this standard). Future editions to this standard may also include
plumulosus, and R. abronius. While standard procedures are
methods for conducting the toxicity tests in smaller chambers
described in this standard, further investigation of certain
with less sediment (Ho et al. 2000 (3), Ferretti et al. 2002 (4)).
issues could aid in the interpretation of test results. Some of
these issues include the effect of shipping on organism
1.6 Procedures outlined in this standard are based primarily
sensitivity, additional performance criteria for organism health,
on procedures described in the USEPA (1994a (1)), USEPA-
sensitivity of various populations of the same test species, and
USACE (2001(2)), Test Method E1706, and Guides E1391,
confirmation of responses in laboratory tests with natural
E1525, E1688, Environment Canada (1992 (5)), DeWitt et al.
benthos populations.
(1992a (6); 1997a (7)), Emery et al. (1997 (8)), and Emery and
Moore (1996 (9)), Swartz et al. (1985 (10)), DeWitt et al. (1989
1.10 General procedures described in this standard might be
(11)), Scott and Redmond (1989 (12)), and Schlekat et al.
useful for conducting tests with other estuarine or marine
(1992 (13)).
organisms (for example, Corophium spp., Grandidierella
japonica, Lepidactylus dytiscus, Streblospio benedicti), al-
1.7 Additional sediment toxicity research and methods de-
though modifications may be necessary. Results of tests, even
velopment are now in progress to (1) refine sediment spiking
those with the same species, using procedures different from
procedures, (2) refine sediment dilution procedures, (3) refine
those described in the test method may not be comparable and
sediment Toxicity Identification Evaluation (TIE) procedures,
using these different procedures may alter bioavailability.
(4) produce additional data on confirmation of responses in
Comparison of results obtained using modified versions of
laboratory tests with natural populations of benthic organisms
these procedures might provide useful information concerning
(that is, field validation studies), and (5) evaluate relative
new concepts and procedures for conducting sediment tests
sensitivity of endpoints measured in 10- and 28-d toxicity tests
with aquatic organisms. If tests are conducted with procedures
using estuarine or marine amphipods. This information will be
different from those described in this test method, additional
described in future editions of this standard.
tests are required to determine comparability of results. Gen-
1.8 Although standard procedures are described in Annex
eral procedures described in this test method might be useful
A2 of this standard for conducting chronic sediment tests with
for conducting tests with other aquatic organisms; however,
L. plumulosus, further investigation of certain issues could aid
modifications may be necessary.
in the interpretation of test results. Some of these issues include
1.11 Selection of Toxicity Testing Organisms:
further investigation to evaluate the relative toxicological
sensitivity of the lethal and sublethal endpoints to a wide 1.11.1 The choice of a test organism has a major influence
variety of chemicals spiked in sediment and to mixtures of on the relevance, success, and interpretation of a test.
chemicals in sediments from contamination gradients in the Furthermore, no one organism is best suited for all sediments.
field (USEPA-USACE 2001 (2)). Additional research is needed The following criteria were considered when selecting test
to evaluate the ability of the lethal and sublethal endpoints to organisms to be described in this standard (Table 1 and Guide
estimate the responses of populations and communities of E1525). Ideally, a test organism should: (1) have a toxicologi-
benthic invertebrates to contaminated sediments. Research is cal database demonstrating relative sensitivity to a range of
E1367 − 03 (2023)
contaminants of interest in sediment, (2) have a database for abdita, E. estuarius, L. plumulosus, and R. abronius must be
interlaboratory comparisons of procedures (for example, developed in order for these and other organisms to be included
in future editions of this standard.
round-robin studies), (3) be in direct contact with sediment, (4)
be readily available from culture or through field collection, (5) 1.11.3 The primary criterion used for selecting L. plumulo-
sus for chronic testing of sediments was that this species is
be easily maintained in the laboratory, (6) be easily identified,
found in both oligohaline and mesohaline regions of estuaries
(7) be ecologically or economically important, (8) have a broad
on the East Coast of the United States and is tolerant to a wide
geographical distribution, be indigenous (either present or
range of sediment grain size distribution (USEPA-USACE
historical) to the site being evaluated, or have a niche similar to
2001 (2), Annex Annex A2). This species is easily cultured in
organisms of concern (for example, similar feeding guild or
the laboratory and has a relatively short generation time (that
behavior to the indigenous organisms), (9) be tolerant of a
is, about 24 d at 23 °C, DeWitt et al. 1992a(6)) that makes this
broad range of sediment physico-chemical characteristics (for
species adaptable to chronic testing (Section 12).
example, grain size), and (10) be compatible with selected
1.11.4 An important consideration in the selection of spe-
exposure methods and endpoints (Guide E1525). Methods
cific species for test method development is the existence of
utilizing selected organisms should also be (11) peer reviewed
information concerning relative sensitivity of the organisms
(for example, journal articles) and (12) confirmed with re-
both to single chemicals and complex mixtures. Several studies
sponses with natural populations of benthic organisms.
have evaluated the sensitivities of A. abdita, E. estuarius, L.
1.11.2 Of these criteria (Table 1), a database demonstrating
plumulosus, or R. abronius, either relative to one another, or to
relative sensitivity to contaminants, contact with sediment,
other commonly tested estuarine or marine species. For
ease of culture in the laboratory or availability for field-
example, the sensitivity of marine amphipods was compared to
collection, ease of handling in the laboratory, tolerance to
other species that were used in generating saltwater Water
varying sediment physico-chemical characteristics, and confir-
Quality Criteria. Seven amphipod genera, including Ampelisca
mation with responses with natural benthic populations were
abdita and Rhepoxynius abronius, were among the test species
the primary criteria used for selecting A. abdita, E. estuarius,
used to generate saltwater Water Quality Criteria for 12
L. plumulosus, and R. abronius for the current edition of this
chemicals. Acute amphipod toxicity data from 4-d water-only
standard for 10-d sediment tests (Annex A1). The species
tests for each of the 12 chemicals was compared to data for (1)
chosen for this method are intimately associated with sediment,
all other species, (2) other benthic species, and (3) other
due to their tube- dwelling or free-burrowing, and sediment
infaunal species. Amphipods were generally of median sensi-
ingesting nature. Amphipods have been used extensively to test
tivity for each comparison. The average percentile rank of
the toxicity of marine, estuarine, and freshwater sediments
amphipods among all species tested was 57 %; among all
(Swartz et al., 1985 (10); DeWitt et al., 1989 (11); Scott and
benthic species, 56 %; and, among all infaunal species, 54 %.
Redmond, 1989 (12); DeWitt et al., 1992a (6); Schlekat et al.,
Thus, amphipods are not uniquely sensitive relative to all
1992 (13)). The selection of test species for this standard
species, benthic species, or even infaunal species (USEPA
followed the consensus of experts in the field of sediment
1994a (1)). Additional research may be warranted to develop
toxicology who participated in a workshop entitled “Testing
tests using species that are consistently more sensitive than
Issues for Freshwater and Marine Sediments”. The workshop
amphipods, thereby offering protection to less sensitive groups.
was sponsored by USEPA Office of Water, Office of Science
1.11.5 Williams et al. (1986 (16)) compared the sensitivity
and Technology, and Office of Research and Development, and
of the R. abronius 10-d whole sediment test, the oyster embryo
was held in Washington, D.C. from 16-18 September 1992
(Crassostrea gigas) 48-h abnormality test, and the bacterium
(USEPA, 1992 (15)). Of the candidate species discussed at the
(Vibrio fisheri) 1-h luminescence inhibition test (that is, the
workshop, A. abdita, E. estuarius, L. plumulosus, and R.
Microtox test) to sediments collected from 46 contaminated
abronius best fulfilled the selection criteria, and presented the
sites in Commencement Bay, WA. Rhepoxynius abronius were
availability of a combination of one estuarine and one marine
exposed to whole sediment, while the oyster and bacterium
species each for both the Atlantic (the estuarine L. plumulosus
tests were conducted with sediment elutriates and extracts,
and the marine A. abdita ) and Pacific (the estuarine E.
respectfully. Microtox was the most sensitive test, with 63 %
estuarius and the marine R. abronius) coasts. Ampelisca abdita
of the sites eliciting significant inhibition of luminescence.
is also native to portions of the Gulf of Mexico and San
Significant mortality of R. abronius was observed in 40 % of
Francisco Bay. Many other organisms that might be appropri-
test sediments, and oyster abnormality occurred in 35 % of
ate for sediment testing do not now meet these selection criteria
sediment elutriates. Complete concordance (that is, sediments
because little emphasis has been placed on developing stan-
that were either toxic or not-toxic in all three tests) was
dardized testing procedures for benthic organisms. For
observed in 41 % of the sediments. Possible sources for the
example, a fifth species, Grandidierella japonica was not
lack of concordance at other sites include interspecific differ-
selected because workshop participants felt that the use of this
ences in sensitivity among test organisms, heterogeneity in
species was not sufficiently broad to warrant standardization of
contaminant types associated with test sediments, and differ-
the method. Environment Canada (1992 (5)) has recommended
ences in routes of exposure inherent in each toxicity test. These
the use of the following amphipod species for sediment toxicity
testing: Amphiporeia virginiana, Corophium volutator, Eo-
haustorius washingtonianus, Foxiphalus xiximeus, and Lep-
Microtox is a trademark of Strategic Diagnostics Inc. 111 Pencader Drive
tocheirus pinguis. A database similar to those available for A. Newark, Delaware 19702-3322.
E1367 − 03 (2023)
results highlight the importance of using multiple assays when using dilutions of sediments collected from Black Rock
performing sediment assessments. Harbor, CT. There was strong agreement among species and
laboratories in the ranking of sediment toxicity and the ability
1.11.6 Several studies have compared the sensitivity of
to discriminate between toxic and non-toxic sediments.
combinations of the four amphipods to sediment contaminants.
1.11.6.4 Hartwell et al. (2000 (23)) evaluated the response
For example, there are several comparisons between A. abdita
of Leptocheirus plumulosus (10-d survival or growth) to the
and R. abronius, between E. estuarius and R. abronius, and
response of the amphipod Lepidactylus dytiscus (10-d survival
between A. abdita and L. plumulosus. There are fewer ex-
or growth), the polychaete Streblospio benedicti (10-d survival
amples of direct comparisons between E. estuarius and L.
or growth), and lettuce germination (Lactuca sativa in 3-d
plumulosus, and no examples comparing L. plumulosus and R.
exposure) and observed that L. plumulosus was relatively
abronius. There is some overlap in relative sensitivity from
insensitive compared to the response of either L. dytiscus or S.
comparison to comparison within each species combination,
benedicti in exposures to 4 sediments with elevated metal
which appears to indicate that all four species are within the
concentrations.
same range of relative sensitivity to contaminated sediments.
1.11.6.1 Word et al. (1989 (17)) compared the sensitivity of 1.11.6.5 Ammonia is a naturally occurring compound in
A. abdita and R. abronius to contaminated sediments in a series marine sediment that results from the degradation of organic
of experiments. Both species were tested at 15 °C. Experiments debris. Interstitial ammonia concentrations in test sediment can
were designed to compare the response of the organism rather range from <1 mg/L to in excess of 400 mg/L (Word et al.,
than to provide a comparison of the sensitivity of the methods 1997 (24)). Some benthic infauna show toxicity to ammonia at
(that is, Ampelisca abdita would normally be tested at 20 °C). concentrations of about 20 mg/L (Kohn et al., 1994 (25)).
Sediments collected from Oakland Harbor, CA, were used for Based on water-only and spiked-sediment experiments with
the comparisons. Twenty-six sediments were tested in one ammonia, threshold limits for test initiation and termination
comparison, while 5 were tested in the other. Analysis of have been established for the L. plumulosus chronic test.
results using Kruskal Wallace rank sum test for both experi- Smaller (younger) individuals are more sensitive to ammonia
than larger (older) individuals (DeWitt et al., 1997a(7), b (26).
ments demonstrated that R. abronius exhibited greater sensi-
tivity to the sediments than A. abdita at 15 °C. Long and Results of a 28-d test indicated that neonates can tolerate very
high levels of pore-water ammonia (>300 mg/L total ammonia)
Buchman (1989 (18)) also compared the sensitivity of A.
for short periods of time with no apparent long-term effects
abdita and R. abronius to sediments from Oakland Harbor, CA.
(Moore et al., 1997 (27)). It is not surprising L. plumulosus has
They also determined that A. abdita showed less sensitivity
a high tolerance for ammonia given that these amphipods are
than R. abronius, but they also showed that A. abdita was less
often found in organic rich sediments in which diagenesis can
sensitive to sediment grain size factors than R. abronius.
result in elevated pore-water ammonia concentrations. Insen-
1.11.6.2 DeWitt et al. (1989 (11)) compared the sensitivity
sitivity to ammonia by L. plumulosus should not be construed
of E. estuarius and R. abronius to sediment spiked with
as an indicator of the sensitivity of the L. plumulosus sediment
fluoranthene and field-collected sediment from industrial wa-
toxicity test to other chemicals of concern.
terways in Puget Sound, WA, in 10-d tests, and to aqueous
cadmium (CdCl ) in a 4-d water-only test. The sensitivity of E. 1.11.7 Limited comparative data is available for concurrent
estuarius was from two (to spiked-spiked sediment) to seven water-only exposures of all four species in single-chemical
(to one Puget Sound, WA, sediment) times less sensitive than tests. Studies that do exist generally show that no one species
R. abronius in sediment tests, and ten times less sensitive to is consistently the most sensitive.
CdCl in the water-only test. These results are supported by the
2 1.11.7.1 The relative sensitivity of the four amphipod spe-
findings of Pastorok and Becker (1990 (19)) who found the
cies to ammonia was determined in ten-d water only toxicity
acute sensitivity of E. estuarius and R. abronius to be generally
tests in order to aid interpretation of results of tests on
comparable to each other, and both were more sensitive than
sediments where this toxicant is present (USEPA 1994a (1)).
Neanthes arenaceodentata (survival and biomass endpoints),
These tests were static exposures that were generally con-
Panope generosa (survival), and Dendraster excentricus (sur-
ducted under conditions (for example, salinity, photoperiod)
vival).
similar to those used for standard 10-d sediment tests. Depar-
tures from standard conditions included the absence of sedi-
1.11.6.3 Leptocheirus plumulosus was as sensitive as the
ment and a test temperature of 20 °C for L. plumulosus, rather
freshwater amphipod Hyalella azteca to an artificially created
than 25 °C as dictated in this standard. Sensitivity to total
gradient of sediment contamination when the latter was accli-
o
ammonia increased with increasing pH for all four species. The
mated to oligohaline salinity (that is, 6 ⁄oo ; McGee et al., 1993
rank sensitivity was R. abronius = A. abdita > E. estuarius > L.
(20)). DeWitt et al. (1992b (21)) compared the sensitivity of L.
plumulosus. A similar study by Kohn et al. (1994 (25)) showed
plumulosus with three other amphipod species, two mollusks,
a similar but slightly different relative sensitivity to ammonia
and one polychaete to highly contaminated sediment collected
with A. abdita > R. abronius = L. plumulosus > E. estuarius.
from Baltimore Harbor, MD, that was serially diluted with
clean sediment. Leptocheirus plumulosus was more sensitive 1.11.7.2 Cadmium chloride has been a common reference
than the amphipods Hyalella azteca and Lepidactylus dytiscus
toxicant for all four species in 4-d exposures. DeWitt et al.
and exhibited equal sensitivity with E. estuarius. Schlekat et al. (1992a (6)) reports the rank sensitivity as R. abronius > A.
(1995 (22)) describe the results of an interlaboratory compari- abdita > L. plumulosus > E. estuarius at a common tempera-
o
son of 10-d tests with A. abdita, L. plumulosus and E. estuarius ture and salinity of 15 °C and 28 ⁄oo . A series of 4-d exposures
E1367 − 03 (2023)
to cadmium that were conducted at species-specific tempera- 1.11.10.1 Data from USEPA Office of Research and Devel-
tures and salinities showed the following rank sensitivity: A. opment’s Environmental Monitoring and Assessment program
abdita = L. plumulosus = R. abronius > E. estuarius (USEPA
were examined to evaluate the relationship between survival of
1994a (1)).
Ampelisca abdita in sediment toxicity tests and the presence of
amphipods, particularly ampeliscids, in field samples. Over
1.11.7.3 Relative species sensitivity frequently varies
among contaminants; consequently, a battery of tests including 200 sediment samples from two years of sampling in the
Virginian Province (Cape Cod, MA, to Cape Henry, VA) were
organisms representing different trophic levels may be needed
to assess sediment quality (Craig, 1984 (28); Williams et al. available for comparing synchronous measurements of A.
1986 (16); Long et al., 1990 (29); Ingersoll et al., 1990 (30); abdita survival in toxicity tests to benthic community enumera-
Burton and Ingersoll, 1994 (31)). For example, Reish (1988
tion. Although species of this genus were among the more
(32)) reported the relative toxicity of six metals (arsenic,
frequently occurring taxa in these samples, ampeliscids were
cadmium, chromium, copper, mercury, and zinc) to totally absent from stations that exhibited A. abdita test
crustaceans, polychaetes, pelecypods, and fishes and concluded
survival <60 % of that in control samples. Additionally, am-
that no one species or group of test organisms was the most
peliscids were found in very low densities at stations with
sensitive to all of the metals.
amphipod test survival between 60 and 80 % (USEPA 1994a
(1)). These data indicate that tests with this species are
1.11.8 The sensitivity of an organism is related to route of
exposure and biochemical response to contaminants. predictive of contaminant effects on sensitive species under
Sediment-dwelling organisms can receive exposure from three natural conditions.
primary sources: interstitial water, sediment particles, and 1.11.10.2 Swartz et al. (1982 (40)) compared sensitivity of
overlying water. Food type, feeding rate, assimilation
R. abronius to sediment collected from sites in Commencement
efficiency, and clearance rate will control the dose of contami- Bay, WA, to benthic community structure at each site. Mortal-
nants from sediment. Benthic invertebrates often selectively
ity of R. abronius was negatively correlated with amphipod
consume different particle sizes (Harkey et al. 1994 (33)) or
density, and phoxocephalid amphipods were ubiquitously ab-
particles with higher organic carbon concentrations which may
sent from the most contaminated areas.
have higher contaminant concentrations. Grazers and other
1.11.10.3 Sediment toxicity to amphipods in 10-d toxicity
collector-gatherers that feed on aufwuchs and detritus may
tests, field contamination, and field abundance of benthic
receive most of their body burden directly from materials
amphipods were examined along a sediment contamination
attached to sediment or from actual sediment ingestion. In
gradient of DDT (Swartz et al. 1994 (39)). Survival of E.
some amphipods (Landrum, 1989 (34)) and clams (Boese et
estuarius and R. abronius in laboratory toxicity tests was
al., 1990 (35)) uptake through the gut can exceed uptake across
positively correlated to abundance of amphipods in the field
the gills for certain hydrophobic compounds. Organisms in
and along with the survival of H. azteca, was negatively
direct contact with sediment may also accumulate contami-
correlated to DDT concentrations. The threshold for 10-d
nants by direct adsorption to the body wall or by absorption
sediment toxicity in laboratory studies was about 300 ug DDT
through the integument (Knezovich et al. 1987 (36)).
(+metabolites)/g organic carbon. The threshold for abundance
1.11.9 Despite the potential complexities in estimating the
of amphipods in the field was about 100 ug DDT
dose that an animal receives from sediment, the toxicity and
(+metabolites)/g organic carbon. Therefore, correlations be-
bioaccumulation of many contaminants in sediment such as
tween toxicity, contamination, and biology indicate that acute
Kepone®, fluoranthene, organochlorines, and metals have been
10-d sediment toxicity tests can provide reliable evidence of
correlated with either the concentration of these chemicals in
biologically adverse sediment contamination in the field.
interstitial water or in the case of non-ionic organic chemicals,
1.11.10.4 As part of a comprehensive sediment quality
concentrations in sediment on an organic carbon normalized
assessment in Baltimore Harbor, MD, McGee et al. (1999 (41))
basis (Di Toro et al. 1990 (37); Di Toro et al. 1991 (38)). The
conducted 10-d toxicity tests with L. plumulosus. Negative
relative importance of whole sediment and interstitial water
relationships were detected between amphipod survival and
routes of exposure depends on the test organism and the
concentrations of select sediment-associated contaminants,
specific contaminant (Knezovich et al. 1987 (36)). Because
whereas a very strong positive association existed between
benthic communities contain a diversity of organisms, many
survival in laboratory exposures and field density of L. plumu-
combinations of exposure routes may be important. Therefore,
losus at test sites. A field validation study of the 10- and 28-d
behavior and feeding habits of a test organism can influence its
L. plumulosus tests by McGee and Fisher (1999 (42)) in
ability to accumulate contaminants from sediment and should
be considered when selecting test organisms for sediment Baltimore Harbor, also indicated good agreement between
testing. acute toxicity, sediment associated contaminants and responses
of the in situ benthic community. In this study, the chronic 28-d
1.11.10 The use of A. abdita, E. estuarius, R. abronius, and
test was less sensitive to sediment contamination than the acute
L. plumulosus in laboratory toxicity studies has been field
10-d test; however, the feeding regime used in this evaluation
validated with natural populations of benthic organisms
is different than the one currently recommended in Annex A2
(Swartz et al. 1994 (39) and Anderson et al. 2001 (14) for E.
estuarius, Swartz et al. 1982 (40) and Anderson et al. 2001 (14) and may have influenced the test results. Field validation
studies with the revised 28-d test outlined in Annex A2 have
for R. abronius, McGee et al. 1999 (41) and McGee and Fisher
1999 (42) for L. plumulosus). not been conducted.
E1367 − 03 (2023)
1.12 Chronic Sediment Methods with Leptocheirus plumu-
Sample Collection, Storage, Manipulation, and 10
Characterization
losus:
Quality Assurance and Quality Control 11
1.12.1 Most standard whole sediment toxicity tests have
Collection, Culturing, and Maintaining Test 12
Organisms
been developed to produce a lethality endpoint (survival/
Calculation 13
mortality) with potential for a sublethal endpoint (reburial) in
Report 14
some species (USEPA 1994a (1), USEPA-USACE 2001 (2)).
Precision and Bias 15
Keywords 16
Methods that measure sublethal effects have not been available
Annexes
or have not been routinely used to evaluate sediment toxicity in
A1. Procedure For Conducting A 10-d Sediment Annex A1
marine or estuarine sediments (Scott and Redmond, 1989 (12); Survival Test With the Amphipods Ampelisca abdita,
Eohaustorius estuarius, Leptocheirus plumulosus,,
Green and Chandler, 1996 (43); Levin et al., 1996 (44); Ciarelli
or Rhepoxynius abronius
et al., 1998 (45); Meador and Rice, 2001 (46)). Most assess-
A2. Procedure For Conducting A Leptocheirus Annex A2
ments of contaminated sediment rely on short-term lethality plumulosus 28-d Sediment For Measuring Sublethal
Effects of Sediment-Associated Contaminants.
tests (for example, ≤10 d; USEPA-USACE, 1991 (47); 1998
References
(48)). Short-term lethality tests are useful in identifying “hot
1.15 This standard does not purport to address all of the
spots” of sediment contamination, but might not be sensitive
safety concerns, if any, associated with its use. It is the
enough to evaluate moderately contaminated areas. However,
responsibility of the user of this standard to establish appro-
sediment quality assessments using sublethal responses of
priate safety, health, and environmental practices and deter-
benthic organisms, such as effects on growth and reproduction,
mine the applicability of regulatory limitations prior to use.
have been used to successfully evaluate moderately contami-
This standard does not purport to address all of the safety
nated areas (Ingersoll et al., 1998 (49); Kemble et al., 1994
concerns, if any, associated with its use. It is the responsibility
(50); McGee et al., 1995 (51); Scott, 1989 (52)). The 28-d
of the user of this standard to establish appropriate safety and
toxicity test with Leptocheirus plumulosus has two sublethal
health practices and determine the applicability of regulatory
endpoints: growth and reproduction. These sublethal endpoints
limitations prior to use.Specific hazard statements are given in
have potential to exhibit a toxic response from chemicals that
Section 8.
otherwise might not cause acute effects or significant mortality
1.16 This international standard was developed in accor-
in a test. Sublethal response to chronic exposure is also
dance with internationally recognized principles on standard-
valuable for population modeling of contaminant effects. These
ization established in the Decision on Principles for the
data can be used for population-level risk assessments of
Development of International Standards, Guides and Recom-
benthic pollutant effects.
mendations issued by the World Trade Organization Technical
1.12.2 An evaluation of the distribution of L. plumulosus in
Barriers to Trade (TBT) Committee.
Chesapeake Bay indicates that its distribution is negatively
correlated with the degree of sediment contamination
2. Referenced Documents
(Pfitzenmeyer, 1975 (53); Reinharz, 1981 (54)). A field vali-
2.1 ASTM Standards:
dation study of the 10- and 28-d L. plumulosus tests by McGee
D1129 Terminology Relating to Water
and Fisher (1999 (42)) in Baltimore Harbor, indicated good
D4447 Guide for Disposal of Laboratory Chemicals and
agreement between acute toxicity, sediment associated con-
Samples
taminants and responses of the in situ benthic community. In
E29 Practice for Using Significant Digits in Test Data to
this study, the chronic 28-d test was less sensitive to sediment
Determine Conformance with Specifications
contamination than the acute 10-d test and therefore had a
E105 Guide for Probability Sampling of Materials
poorer association between sediment contaminants and benthic
E122 Practice for Calculating Sample Size to Estimate, With
community health. It should be noted that the feeding regime
Specified Precision, the Average for a Characteristic of a
used in this evaluation is different than the one currently
Lot or Process
recommended in Annex A2 and may have influenced the test
E141 Practice for Acceptance of Evidence Based on the
results. Field validation studies with the revised 28-d test have
Results of Probability Sampling
not been conducted.
E177 Practice for Use of the Terms Precision and Bias in
1.13 Limitations—While some safety considerations are
ASTM Test Methods
included in this standard, it is beyond the scope of this standard E178 Practice for Dealing With Outlying Observations
to encompass all safety requirements necessary to conduct E456 Terminology Relating to Quality and Statistics
sediment tests. E691 Practice for Conducting an Interlaboratory Study to
Determine the Precision of a Test Method
1.14 This standard is arranged as follows:
E729 Guide for Conducting Acute Toxicity Tests on Test
Section
Materials with Fishes, Macroinvertebrates, and Amphib-
Referenced Documents 2
ians
Terminology 3
Summary of Standard 4
Significance and Use 5
Interferences 6
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Reagents and Materials 7
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
Hazards 8
Standards volume information, refer to the standard’s Document Summary page on
Facilities, Equipment, and Supplies 9
the ASTM website.
E1367 − 03 (2023)
E943 Terminology Relating to Biological Effects and Envi- 3.3.3 contaminated sediment, n—sediment containing
ronmental Fate (Withdrawn 2023) chemical substances at concentrations that pose a known or
E1241 Guide for Conducting Early Life-Stage Toxicity Tests suspected threat to environmental or human health.
with Fishes
3.3.4 control sediment, n—a sediment that is essentially free
E1325 Terminology Relating to Design of Experiments
of contaminants and is used routinely to assess the acceptabil-
E1391 Guide for Collection, Storage, Characterization, and
ity of a test. Any contaminants in control sediment may
Manipulation of Sediments for Toxicological Testing and
originate from the global spread of pollutants and does not
for Selection of Samplers Used to Collect Benthic Inver-
reflect any substantial input from local or non-point sources.
tebrates
Comparing test sediments to control sediments is a measure of
E1402 Guide for Sampling Design
the toxicity of a test sediment beyond inevitable background
E1525 Guide for Designing Biological Tests with Sediments
contamination.
E1611 Guide for Conducting Sediment Toxicity Tests with
3.3.5 EC50, n—a statistically or graphically estimated con-
Polychaetous Annelids
centration that is expected to cause one or more specified
E1688 Guide for Determination of the Bioaccumulation of
effects in 50 % of a group of organisms under specified
Sediment-Associated Contaminants by Benthic Inverte-
conditions.
brates
3.3.6 formulated sediment, n—mixtures of materials used to
E1706 Test Method for Measuring the Toxicity of Sediment-
mimic the physical components of a natural sediment.
Associated Contaminants with Freshwater Invertebrates
E1847 Practice for Statistical Analysis of Toxicity Tests
3.3.7 IC50, n—a point estimate of the toxicant concentration
Conducted Under ASTM Guidelines (Withdrawn 2022)
that would cause a 50 % reduction in a non-quantal measure-
E1850 Guide for Selection of Resident Species as Test
ment such as fecundity or growth.
Organisms for Aquatic and Sediment Toxicity Tests
3.3.8 interstitial water or pore water, n— water occupying
IEEE/ASTM SI 10 American National Standard for Use of
space between sediment or soil particles.
the International System of Units (SI): The Modern Metric
3.3.9 LC50, n—a statistically or graphically estimated con-
System
centration that is expected to be lethal to 50 % of a group of
organisms under specified conditions.
3. Terminology
3.3.10 lowest-observable-effect concentration (LOEC),
3.1 The words “must,” “should,” “may,” “can,” and “might”
n—in a toxicity test, the lowest tested concentration of a
have very specific meanings in this standard. “Must“ is used to
material at which organisms were adversely affected compared
express an absolute requirement, that is, to state that a test
to control organisms as determined by statistical hypothesis
ought to be designed to satisfy the specified conditions, unless
tests-should be accompanied by a description of the statistical
the purpose of the test requires a different design. “Must” is
tests and alternative hypotheses, levels of significance, and
used only in connection with the factors that relate directly to
measures of performance, for example, survival, growth,
the acceptability of a test. “Should” is used to state that the
reproduction, or development-and must be above any other
specified condition is recommended and ought to be met if
concentration not producing statistically significant adverse
possible. Although the violation of one “should” is rarely a
effects.
serious matter, violation of several will often render the results
questionable. Terms such as “is desirable,” “is often desirable,”
3.3.11 no-observable-effect concentration (NOEC), n—in a
and “might be desirable” are used in connection with less toxicity test, the highest tested concentration of a material at
important factors. “May” is used to mean “is (are) allowed to,”
which organisms did as well as control organisms as deter-
“can” is used to mean “is (are) able to,” and “might” is used to mined by statistical hypothesis tests-should be accompanied by
mean “could possibly.” Thus, the classic distinction between
a description of the statistical tests and alternative hypotheses,
“may” and “can” is preserved, and “might” is never used as a levels of significance, and measures of performance, for
synonym for either “may” or “can.”
example, survival, growth, reproduction, or development-and
must be below any other concentration producing statistically
3.2 Definitions—For definitions of other terms used in this
significant adverse effects.
test method, refer to Guides E729 and E1241 and Terminology
E943 and D1129. For an explanation of units and symbols, 3.3.12 overlying water, n—the water placed over sediment
refer to IEEE/ASTM SI 10IEEE/ASTM SI 10. in a test chamber during a test.
3.3.13 reference sediment, n—a whole sediment near an
3.3 Definitions of Terms Specific to This Standard:
area of concern used to assess sediment conditions exclusive of
3.3.1 clean, n—denotes a sediment or water that does not
material(s) of interest. The reference sediment may be used as
contain concentrations of test materials which cause apparent
an indicator of localized sediment conditions exclusive of the
stress to the test organisms or reduce their survival.
specific pollutant input of concern. Such sediment would be
3.3.2 concentration, n—the ratio of weight or volume of test
collected near the site of concern and would represent the
material(s) to the weight or volume of sediment.
background conditions resulting from any localized pollutant
inputs as well as global pollutant input. This is the manner in
which reference sediment is used in dredge material evalua-
The last approved version of this historical standard is referenced on
www.astm.org. tions.
E1367 − 03 (2023)
3.3.14 reference-toxicity test, n—a test conducted with termination. Growth rate is calculated as the mean dry weight
reagent-grade reference chemical to assess the sensitivity of the gain per day per adult amphipod surviving at test termination.
test organisms. Deviations outside an established normal range Reproduction is calculated as the number of offspring per
may indicate a change in the sensitivity of the test organism surviving adult. This test is applicable for use with sediment
o o
population. Reference-toxicity tests are most often performed having pore-water salinity ranging from 1 ⁄oo to 35 ⁄oo .
in the absence of sediment. Typically, endpoint selection for new toxicity tests is generally
guided by methodologies for related toxicity tests (Gray et al.,
3.3.15 sediment, n—particulate material that usually lies
1998 (55)). Sediment toxicity tests using macroinvertebrates
below water. Formulated particulate material that is intended to
of
...