ASTM E1192-97(2008)
(Guide)Standard Guide for Conducting Acute Toxicity Tests on Aqueous Ambient Samples and Effluents with Fishes, Macroinvertebrates, and Amphibians
Standard Guide for Conducting Acute Toxicity Tests on Aqueous Ambient Samples and Effluents with Fishes, Macroinvertebrates, and Amphibians
ABSTRACT
This guide covers procedures for obtaining laboratory data concerning the adverse effects of aqueous ambient samples and effluents on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, during a short-term exposure, depending on the species, using the static, renewal, and flow-through techniques. These procedures will probably be useful for conducting acute toxicity tests on aqueous effluents with many other aquatic species, although modifications might be necessary. Static tests might not be applicable to effluents that have a high oxygen demand, or contain materials that (1) are highly volatile, (2) are rapidly biologically or chemically transformed in aqueous solutions, or (3) are removed from test solutions in substantial quantities by the test chambers or organisms during the test. Results of acute toxicity tests should usually be reported in terms of a median lethal concentration (LC50) or median effective concentration (EC50). An acute toxicity test does not provide information about whether delayed effects will occur. Specified requirements involving the following are detailed: (1) hazards; (2) apparatus: facilities, special requirements, construction materials, metering system, test chambers, cleaning, and acceptability; (3) dilution water requirements, source, treatment, and characterization; (4) effluent sampling point, collection, preservation, treatment, and test concentrations; (5) test organism species, age, source, care and handling, feeding, disease treatment, holding, acclimation, and quality; (6) procedure: experimental design, dissolved oxygen, temperature, loading, beginning the test, feeding, duration of test, biological data, and other measurements; (7) analytical methodology; (8) acceptability of test; (9) calculation of results; and (1) report of results.
SIGNIFICANCE AND USE
An acute effluent toxicity test is conducted to obtain information concerning the immediate effects on test organisms of a short-term exposure to an effluent under specific experimental conditions. One can directly examine acute effects of complex mixtures of chemicals as occurs in effluents and some ambient waters. Acute effluent toxicity tests can be used to evaluate the potential for designated-use or aquatic life imperiment in the receiving stream, lake, or estuary. An acute toxicity test does not provide information about whether delayed effects will occur, although a post-exposure observation period, with appropriate feeding if necessary, might provide such information.
Results of acute effluent tests might be used to predict acute effects likely to occur on aquatic organisms in field situations as a result of exposure under comparable conditions, except that (1) motile organisms might avoid exposure when possible, (2) toxicity to benthic species might be dependent on sorption or settling of components of the effluent onto the substrate, and (3) the effluent might physically or chemically interact with the receiving water.
Results of acute effluent tests might be used to compare the acute sensitivities of different species and the acute toxicities of different effluents, and to study the effects of various environmental factors on results of such tests.
Acute tests are usually the first step in evaluating the effects of an effluent on aquatic organisms.
Results of acute effluent tests will depend on the temperature, composition of the dilution water, condition of the test organisms, exposure technique, and other factors.
SCOPE
1.1 This guide covers procedures for obtaining laboratory data concerning the adverse effects of an aqueous effluent on certain species of freshwater and saltwater fishes, macroinvertebrates, and amphibians, usually during 2 to 4-day exposures, depending on the species, using the static, renewal, and flow-through techniques. These procedures will probably be useful for conducting acute toxicity tests on aqueous effluents with many...
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Designation: E1192 − 97(Reapproved 2008)
Standard Guide for
Conducting Acute Toxicity Tests on Aqueous Ambient
Samples and Effluents with Fishes, Macroinvertebrates, and
Amphibians
This standard is issued under the fixed designation E1192; 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 and effluent are maintained at desired levels and degradation
and metabolic products are removed. Static tests might not be
1.1 This guide covers procedures for obtaining laboratory
applicable to effluents that have a high oxygen demand, or
data concerning the adverse effects of an aqueous effluent on
contain materials that (1) are highly volatile, (2) are rapidly
certain species of freshwater and saltwater fishes,
biologicallyorchemicallytransformedinaqueoussolutions,or
macroinvertebrates, and amphibians, usually during 2 to 4-day
(3) are removed from test solutions in substantial quantities by
exposures, depending on the species, using the static, renewal,
the test chambers or organisms during the test. Flow-through
and flow-through techniques. These procedures will probably
testsaregenerallypreferabletorenewaltests,althoughinsome
be useful for conducting acute toxicity tests on aqueous
situations a renewal test might be more cost-effective than a
effluents with many other aquatic species, although modifica-
flow-through test.
tions might be necessary.
1.5 In the development of these procedures, an attempt was
1.2 Other modifications of these procedures might be justi-
made to balance scientific and practical considerations and to
fied by special needs or circumstances.Although using appro-
ensure that the results will be sufficiently accurate and precise
priate procedures is more important than following prescribed
for the applications for which they are commonly used. A
procedures,resultsoftestsconductedusingunusualprocedures
majorconsiderationwasthatthecommonusesoftheresultsof
are not likely to be comparable to results of many other tests.
acute tests on effluents do not require or justify stricter
Comparisonofresultsobtainedusingmodifiedandunmodified
requirements than those set forth in this guide. Although the
versions of these procedures might provide useful information
tests may be improved by using more organisms, longer
concerning new concepts and procedures for conducting acute
acclimation times, and so forth, the requirements presented in
toxicity tests on aqueous effluents.
this guide should usually be sufficient.
1.3 This guide is based in large part on Guide E729. The
1.6 Resultsofacutetoxicitytestsshouldusuallybereported
majordifferencesbetweenthetwoguidesare(1)themaximum
in terms of a median lethal concentration (LC50) or median
test concentration is 100% effluent or ambient sample, (2)
effective concentration (EC50). In some situations, it might be
testing is not chemical specific, and (3) the holding time of
necessaryonlytodeterminewhetheraspecificconcentrationis
effluent and ambient samples is often considerably less than
acutely toxic to the test species or whether the LC50 or EC50
thatforchemicalsandothertestmaterials.Becausethesample
is above or below a specific concentration.
isoftenacomplexmixtureofchemicals,analyticaltestscannot
generally be used to confirm exposure concentrations.
1.7 This guide is arranged as follows:
Section
1.4 Selection of the technique to be used in a specific
situation will depend upon the needs of the investigator and
Referenced Documents 2
upon available resources. Static tests provide the most easily
Terminology 3
Summary of Guide 4
obtained measure of acute toxicity, but should not last longer
Significance and Use 5
than48h.Renewalandflow-throughtestsmaylastlongerthan
Hazards 7
48 h because the pH and concentrations of dissolved oxygen
Apparatus 6
Facilities 6.1
Special Requirements 6.2
Construction Materials 6.3
Metering System 6.4
ThisguideisunderthejurisdictionofASTMCommitteeE50onEnvironmental
Test Chambers 6.5
Assessment, Risk Management and CorrectiveAction and is the direct responsibil-
Cleaning 6.6
ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.
Acceptability 6.7
Current edition approved Feb. 1, 2008. Published February 2008. Originally
Dilution Water 8
approved in 1988. Last previous edition approved in 2003 as E1192–97(2003).
Requirements 8.1
DOI: 10.1520/E1192-97R08.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1192 − 97 (2008)
express an absolute requirement, that is, to state that the test
Source 8.2
Treatment 8.3
ought to be designed to satisfy the specified condition, unless
Characterization 8.4
the purpose of the test requires a different design. “Must” is
Effluent 9
only used in connection with factors that directly relate to the
Sampling Point 9.1
Collection 9.2
acceptability of the test (see 13.1). “Should” is used to state
Preservation 9.3
that the specified condition is recommended and ought to be
Treatment 9.4
met if possible.Although violation of one “should” is rarely a
Test Concentration(s) 9.5
Test Organisms 10
serious matter, violation of several will often render the results
Species 10.1
questionable.Termssuchas“isdesirable,”“isoftendesirable,”
Age 10.2
Source 10.3 and “might be desirable” are used in connection with less
Care and Handling 10.4
importantfactors.“May”isusedtomean“is(are)allowedto,”
Feeding 10.5
“can”isusedtomean“is(are)ableto,”and“might”isusedto
Disease Treatment 10.6
Holding 10.7 mean “could possibly.” Thus the classic distinction between
Acclimation 10.8
“may” and “can” is preserved, and “might” is never used as a
Quality 10.9
synonym for either “may” or “can.”
Procedure 11
Experimental Design 11.1
3.2 The term “effluents” refers to aqueous discharges regu-
Dissolved Oxygen 11.2
lated under the National Pollutant Discharge Elimination
Temperature 11.3
Loading 11.4
System (NPDES) collected at the sampling point specified in
Beginning the Test 11.5
the NPDES permit.
Feeding 11.6
Duration of Test 11.7
3.3 The term “ambient samples” refers to water samples
Biological Data 11.8
collected from the environment. Examples include surface
Other Measurements 11.9
waters, storm waters, leachates, and ground water.
Analytical Methodology 12
Acceptability of Test 13
3.4 Fordefinitionsofothertermsusedinthisguide,referto
Calculation or Results 14
Report 15 Guide E729 and Terminology E943. For an explanation of
units and symbols, refer to IEEE/ASTM SI 10.
1.8 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
4. Summary of Guide
responsibility of the user of this standard to establish appro-
4.1 Ineachoftwoormoretreatments,testorganismsofone
priate safety and health practices and determine the applica-
species are maintained for 2 to 8 days in one or more test
bility of regulatory limitations prior to use. Specific hazard
chambers. In each of the one or more control treatments, the
statements are given in Section 7.
organismsaremaintainedindilutionwatertowhichnoeffluent
has been added in order to provide (1) a measure of the
2. Referenced Documents
acceptability of the test by giving an indication of the quality
2.1 ASTM Standards:
of the test organisms and the suitability of the dilution water,
E724Guide for Conducting Static Acute Toxicity Tests
test conditions, handling procedures, and so forth, and (2) the
Starting with Embryos of Four Species of Saltwater
basis for interpreting data obtained from the other treatments.
Bivalve Molluscs
In each of the one or more other treatments, the organisms are
E729Guide for Conducting Acute Toxicity Tests on Test
maintained in dilution water to which a selected concentration
Materials with Fishes, Macroinvertebrates, and Amphib-
of effluent has been added. Data on effects on the organisms in
ians
each test chamber are usually obtained periodically during the
E943Terminology Relating to Biological Effects and Envi-
test and analyzed to determine LC50s or EC50s for various
ronmental Fate
lengths of exposure.
E1203Practice for Using Brine Shrimp Nauplii as Food for
Test Animals in Aquatic Toxicology (Withdrawn 2013)
5. Significance and Use
E1604Guide for Behavioral Testing in Aquatic Toxicology
5.1 An acute effluent toxicity test is conducted to obtain
IEEE/ASTM SI 10 American National Standard for Use of
information concerning the immediate effects on test organ-
theInternationalSystemofUnits(SI):TheModernMetric
isms of a short-term exposure to an effluent under specific
System
experimental conditions. One can directly examine acute
effects of complex mixtures of chemicals as occurs in effluents
3. Terminology
and some ambient waters. Acute effluent toxicity tests can be
3.1 Thewords“must,”“should,”“may,”“can,”and“might”
used to evaluate the potential for designated-use or aquatic life
have very specific meanings in this guide. “Must” is used to
imperiment in the receiving stream, lake, or estuary.An acute
toxicity test does not provide information about whether
delayed effects will occur, although a post-exposure observa-
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
tion period, with appropriate feeding if necessary, might
contactASTM Customer Service at service@astm.org. ForAnnual Book ofASTM
Standards volume information, refer to the standard’s Document Summary page on
provide such information.
theASTM website.
5.2 Results of acute effluent tests might be used to predict
The last approved version of this historical standard is referenced on
www.astm.org. acute effects likely to occur on aquatic organisms in field
E1192 − 97 (2008)
situationsasaresultofexposureundercomparableconditions, 6.3 Construction Materials—Equipment and facilities that
except that (1) motile organisms might avoid exposure when contacteffluentsamples,testsolutions,oranywaterintowhich
possible, (2) toxicity to benthic species might be dependent on test organisms will be placed should not contain substances
sorption or settling of components of the effluent onto the that can be leached or dissolved by aqueous solutions in
amounts that adversely affect aquatic organisms. In addition,
substrate, and (3) the effluent might physically or chemically
interact with the receiving water. equipment and facilities that contact effluent samples or test
solutions should be chosen to minimize sorption of effluent
5.3 Results of acute effluent tests might be used to compare
componentsfromwater.Glass,Type316stainlesssteel,nylon,
the acute sensitivities of different species and the acute
and fluorocarbon plastics should be used whenever possible to
toxicities of different effluents, and to study the effects of
minimize dissolution, leaching, and sorption, except that stain-
various environmental factors on results of such tests.
less steel should not be used in tests on metals in salt water.
Concrete and rigid plastics may be used for holding,
5.4 Acute tests are usually the first step in evaluating the
acclimation,andculturetanksandinthewatersupply,butthey
effects of an effluent on aquatic organisms.
should be soaked, preferably in flowing dilution water, for a
5.5 Results of acute effluent tests will depend on the
weekormorebeforeuse (9).Castironpipeshouldnotbeused
temperature,compositionofthedilutionwater,conditionofthe
with salt water and probably should not be used in a
test organisms, exposure technique, and other factors.
freshwater-supply system because colloidal iron will be added
tothedilutionwaterandstrainerswillbeneededtoremoverust
6. Apparatus
particles. A specially designed system is usually necessary to
obtainsaltwaterfromanaturalwatersource(seeGuideE729).
6.1 Facilities—Although some small organisms can be held
Brass, copper, lead, galvanized metal, and natural rubber
and acclimated in static or renewal systems, most organisms
should not contact dilution water, effluent, or test solutions
are held, acclimated, and cultured in flow-through systems.
before or during the test. Items made of neoprene rubber or
Test chambers should be in a constant-temperature room,
other materials not mentioned above should not be used unless
incubator, or recirculating water bath. A dilution-water tank,
it has been shown that either (1) unfed individuals of a
which may be used to store receiving water, or a headbox is
sensitive aquatic species (see 8.2.3) do not show more signs of
oftenelevatedsodilutionwatercanbegravity-fedintoholding
stress, such as discoloration, unusual behavior, or death, when
andacclimationtanksandtestchambers.Pumpsareoftenused
held for at least 48 h in static dilution water in which the item
to deliver dilution water and effluent to headboxes and tanks.
is soaking than when held in static dilution water that does not
Strainers and air traps should be included in the water supply.
contain the item, or (2) their use will not adversely affect
Headboxes and holding, acclimation, culture, and dilution-
survival, growth, or reproduction of a sensitive species.
water tanks should be equipped for temperature control and
aeration (see 8.3). Air used for aeration should be free of
6.4 Metering System:
fumes, oil, and water; filters to remove oil and water are
6.4.1 For flow-through tests, the metering system should be
desirable. Filtration of air through a 0.22 µm bacterial filter
designed to accommodate the type and concentration(s) of the
might be desirable (5). The facility should be well ventilated
effluent and the necessary flow rates of test solutions. The
andfreeoffumes.Tofurtherreducethepossibilityofcontami-
system should mix the effluent with the dilution water imme-
nation by components of the effluent and other substances,
diately before they enter the test chambers and reproducibly
especiallyvolatileones,holding,acclimation,andculturetanks
(see 6.4.4) supply the selected concentration(s) of effluent (see
should not be in a room in which toxicity tests are conducted,
9.5).Variousmeteringsystems,usingdifferentcombinationsof
effluent is stored, test solutions are prepared, or equipment is
syringes, dipping birds, siphons, pumps, saturators, solenoids,
cleaned. During holding, acclimation, culture, and testing,
valves,andsoforth,havebeenusedsuccessfullytocontrolthe
organisms should be shielded from disturbances with curtains
concentrations of effluent in, and the flow rates of, test
or partitions to prevent unnecessary stress. A timing device
solutions (see Guide E729).
should be used to provide a 16-h light and 8-h dark photope-
6.4.2 The f
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