1、Designation: E1192 97 (Reapproved 2014)Standard Guide forConducting Acute Toxicity Tests on Aqueous AmbientSamples and Effluents with Fishes, Macroinvertebrates, andAmphibians1This standard is issued under the fixed designation E1192; the number immediately following the designation indicates the ye
2、ar 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 for obtaining laboratoryd
3、ata concerning the adverse effects of an aqueous effluent oncertain species of freshwater and saltwater fishes,macroinvertebrates, and amphibians, usually during 2 to 4-dayexposures, depending on the species, using the static, renewal,and flow-through techniques. These procedures will probablybe use
4、ful for conducting acute toxicity tests on aqueouseffluents with many other aquatic species, although modifica-tions might be necessary.1.2 Other modifications of these procedures might be justi-fied by special needs or circumstances. Although using appro-priate procedures is more important than fol
5、lowing prescribedprocedures, results of tests conducted using unusual proceduresare not likely to be comparable to results of many other tests.Comparison of results obtained using modified and unmodifiedversions of these procedures might provide useful informationconcerning new concepts and procedur
6、es for conducting acutetoxicity tests on aqueous effluents.1.3 This guide is based in large part on Guide E729. Themajor differences between the two guides are (1) the maximumtest concentration is 100 % effluent or ambient sample, (2)testing is not chemical specific, and (3) the holding time ofefflu
7、ent and ambient samples is often considerably less thanthat for chemicals and other test materials. Because the sampleis often a complex mixture of chemicals, analytical tests cannotgenerally be used to confirm exposure concentrations.1.4 Selection of the technique to be used in a specificsituation
8、will depend upon the needs of the investigator andupon available resources. Static tests provide the most easilyobtained measure of acute toxicity, but should not last longerthan 48 h. Renewal and flow-through tests may last longer than48 h because the pH and concentrations of dissolved oxygenand ef
9、fluent are maintained at desired levels and degradationand metabolic products are removed. Static tests might not beapplicable to effluents that have a high oxygen demand, orcontain materials that (1) are highly volatile, (2) are rapidlybiologically or chemically transformed in aqueous solutions, or
10、(3) are removed from test solutions in substantial quantities bythe test chambers or organisms during the test. Flow-throughtests are generally preferable to renewal tests, although in somesituations a renewal test might be more cost-effective than aflow-through test.1.5 In the development of these
11、procedures, an attempt wasmade to balance scientific and practical considerations and toensure that the results will be sufficiently accurate and precisefor the applications for which they are commonly used. Amajor consideration was that the common uses of the results ofacute tests on effluents do n
12、ot require or justify stricterrequirements than those set forth in this guide. Although thetests may be improved by using more organisms, longeracclimation times, and so forth, the requirements presented inthis guide should usually be sufficient.1.6 Results of acute toxicity tests should usually be
13、reportedin terms of a median lethal concentration (LC50) or medianeffective concentration (EC50). In some situations, it might benecessary only to determine whether a specific concentration isacutely toxic to the test species or whether the LC50 or EC50is above or below a specific concentration.1.7
14、This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Hazards 7Apparatus 6Facilities 6.1Special Requirements 6.2Construction Materials 6.3Metering System 6.4Test Chambers 6.5Cleaning 6.6Acceptability 6.7Dilution Water 8Requirements 8.11Th
15、is guide is under the jurisdiction ofASTM Committee E50 on EnvironmentalAssessment, Risk Management and Corrective Action and is the direct responsibil-ity of Subcommittee E50.47 on Biological Effects and Environmental Fate.Current edition approved Oct. 1, 2014. Published December 2014. Originallyap
16、proved in 1988. Last previous edition approved in 2008 as E1192 97(2008).DOI: 10.1520/E1192-97R14.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1Source 8.2Treatment 8.3Characterization 8.4Effluent 9Sampling Point 9.1Collection 9.2Pre
17、servation 9.3Treatment 9.4Test Concentration(s) 9.5Test Organisms 10Species 10.1Age 10.2Source 10.3Care and Handling 10.4Feeding 10.5Disease Treatment 10.6Holding 10.7Acclimation 10.8Quality 10.9Procedure 11Experimental Design 11.1Dissolved Oxygen 11.2Temperature 11.3Loading 11.4Beginning the Test 1
18、1.5Feeding 11.6Duration of Test 11.7Biological Data 11.8Other Measurements 11.9Analytical Methodology 12Acceptability of Test 13Calculation or Results 14Report 151.8 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the u
19、ser of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use. Specific hazardstatements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:2E724 Guide for Conducting Static Acute Toxicity TestsStartin
20、g with Embryos of Four Species of SaltwaterBivalve MolluscsE729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE943 Terminology Relating to Biological Effects and Envi-ronmental FateE1203 Practice for Using Brine Shrimp Nauplii as Food forT
21、est Animals in Aquatic Toxicology (Withdrawn 2013)3E1604 Guide for Behavioral Testing in Aquatic ToxicologyIEEE/ASTM SI 10 American National Standard for Use ofthe International System of Units (SI):The Modern MetricSystem3. Terminology3.1 The words “must,” “should,” “may,” “can,” and “might”have ve
22、ry specific meanings in this guide. “Must” is used toexpress an absolute requirement, that is, to state that the testought to be designed to satisfy the specified condition, unlessthe purpose of the test requires a different design. “Must” isonly used in connection with factors that directly relate
23、to theacceptability of the test (see 13.1). “Should” is used to statethat the specified condition is recommended and ought to bemet if possible. Although violation of one “should” is rarely aserious matter, violation of several will often render the resultsquestionable. Terms such as “is desirable,”
24、 “is often desirable,”and “might be desirable” are used in connection with lessimportant factors. “May” is used to mean “is (are) allowed to,”“can” is used to mean “is (are) able to,” and “might” is used tomean “could possibly.” Thus the classic distinction between“may” and “can” is preserved, and “
25、might” is never used as asynonym for either “may” or “can.”3.2 The term “effluents” refers to aqueous discharges regu-lated under the National Pollutant Discharge EliminationSystem (NPDES) collected at the sampling point specified inthe NPDES permit.3.3 The term “ambient samples” refers to water sam
26、plescollected from the environment. Examples include surfacewaters, storm waters, leachates, and ground water.3.4 For definitions of other terms used in this guide, refer toGuide E729 and Terminology E943. For an explanation ofunits and symbols, refer to IEEE/ASTM SI 10.4. Summary of Guide4.1 In eac
27、h of two or more treatments, test organisms of onespecies are maintained for 2 to 8 days in one or more testchambers. In each of the one or more control treatments, theorganisms are maintained in dilution water to which no effluenthas been added in order to provide (1) a measure of theacceptability
28、of the test by giving an indication of the qualityof the test organisms and the suitability of the dilution water,test conditions, handling procedures, and so forth, and (2) thebasis for interpreting data obtained from the other treatments.In each of the one or more other treatments, the organisms a
29、remaintained in dilution water to which a selected concentrationof effluent has been added. Data on effects on the organisms ineach test chamber are usually obtained periodically during thetest and analyzed to determine LC50s or EC50s for variouslengths of exposure.5. Significance and Use5.1 An acut
30、e effluent toxicity test is conducted to obtaininformation concerning the immediate effects on test organ-isms of a short-term exposure to an effluent under specificexperimental conditions. One can directly examine acuteeffects of complex mixtures of chemicals as occurs in effluentsand some ambient
31、waters. Acute effluent toxicity tests can beused to evaluate the potential for designated-use or aquatic lifeimperiment in the receiving stream, lake, or estuary. An acutetoxicity test does not provide information about whetherdelayed effects will occur, although a post-exposure observa-tion period,
32、 with appropriate feeding if necessary, mightprovide such information.5.2 Results of acute effluent tests might be used to predictacute effects likely to occur on aquatic organisms in field2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at servic
33、eastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3The last approved version of this historical standard is referenced onwww.astm.org.E1192 97 (2014)2situations as a result of exposure under comparable conditions,except th
34、at (1) motile organisms might avoid exposure whenpossible, (2) toxicity to benthic species might be dependent onsorption or settling of components of the effluent onto thesubstrate, and (3) the effluent might physically or chemicallyinteract with the receiving water.5.3 Results of acute effluent tes
35、ts might be used to comparethe acute sensitivities of different species and the acutetoxicities of different effluents, and to study the effects ofvarious environmental factors on results of such tests.5.4 Acute tests are usually the first step in evaluating theeffects of an effluent on aquatic orga
36、nisms.5.5 Results of acute effluent tests will depend on thetemperature, composition of the dilution water, condition of thetest organisms, exposure technique, and other factors.6. Apparatus6.1 FacilitiesAlthough some small organisms can be heldand acclimated in static or renewal systems, most organ
37、ismsare held, acclimated, and cultured in flow-through systems.Test chambers should be in a constant-temperature room,incubator, or recirculating water bath. A dilution-water tank,which may be used to store receiving water, or a headbox isoften elevated so dilution water can be gravity-fed into hold
38、ingand acclimation tanks and test chambers. Pumps are often usedto deliver dilution water and effluent to headboxes and tanks.Strainers and air traps should be included in the water supply.Headboxes and holding, acclimation, culture, and dilution-water tanks should be equipped for temperature contro
39、l andaeration (see 8.3). Air used for aeration should be free offumes, oil, and water; filters to remove oil and water aredesirable. Filtration of air through a 0.22 m bacterial filtermight be desirable (1). The facility should be well ventilatedand free of fumes. To further reduce the possibility o
40、f contami-nation by components of the effluent and other substances,especially volatile ones, holding, acclimation, and culture tanksshould not be in a room in which toxicity tests are conducted,effluent is stored, test solutions are prepared, or equipment iscleaned. During holding, acclimation, cul
41、ture, and testing,organisms should be shielded from disturbances with curtainsor partitions to prevent unnecessary stress. A timing deviceshould be used to provide a 16-h light and 8-h dark photope-riod.A15 to 30-min transition period (2) when the lights go onmight be desirable to reduce the possibi
42、lity of organisms beingstressed by large, sudden increases in light intensity. A transi-tion period when the lights go off might also be desirable.6.2 Special RequirementsSome organisms require specialconditions during holding, acclimation, and testing. Forexample, burrowing mayfly nymphs should be
43、provided asubstrate suitable for burrowing (3); immature stream insectsshould be in a current (4); and crabs, shrimp, and bottom-dwelling fish should be provided a silica-sand substrate.Because cannibalism might occur among many species ofdecapod crustaceans, the claws of crabs and crayfish should b
44、ebanded, or the individuals should be physically isolated bymeans of screened compartments.6.3 Construction MaterialsEquipment and facilities thatcontact effluent samples, test solutions, or any water into whichtest organisms will be placed should not contain substancesthat can be leached or dissolv
45、ed by aqueous solutions inamounts that adversely affect aquatic organisms. In addition,equipment and facilities that contact effluent samples or testsolutions should be chosen to minimize sorption of effluentcomponents from water. Glass, Type 316 stainless steel, nylon,and fluorocarbon plastics shou
46、ld be used whenever possible tominimize dissolution, leaching, and sorption, except that stain-less steel should not be used in tests on metals in salt water.Concrete and rigid plastics may be used for holding,acclimation, and culture tanks and in the water supply, but theyshould be soaked, preferab
47、ly in flowing dilution water, for aweek or more before use (5). Cast iron pipe should not be usedwith salt water and probably should not be used in afreshwater-supply system because colloidal iron will be addedto the dilution water and strainers will be needed to remove rustparticles. A specially de
48、signed system is usually necessary toobtain salt water from a natural water source (see Guide E729).Brass, copper, lead, galvanized metal, and natural rubbershould not contact dilution water, effluent, or test solutionsbefore or during the test. Items made of neoprene rubber orother materials not me
49、ntioned above should not be used unlessit has been shown that either (1) unfed individuals of asensitive aquatic species (see 8.2.3) do not show more signs ofstress, such as discoloration, unusual behavior, or death, whenheld for at least 48 h in static dilution water in which the itemis soaking than when held in static dilution water that does notcontain the item, or (2) their use will not adversely affectsurvival, growth, or reproduction of a sensitive species.6.4 Metering System:6.4.1 For flow-through tests, the metering system should bedesigned to accommodate the type and co
