1、Designation: E 1241 05Standard Guide forConducting Early Life-Stage Toxicity Tests with Fishes1This standard is issued under the fixed designation E 1241; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A
2、 number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This guide describes procedures for obtaining laboratorydata concerning the adverse effects of a test material added todilution water
3、but not to foodon certain species of fresh-water and saltwater fishes during 28 to 120-day (depending onspecies) continuous exposure, beginning before hatch andending after hatch, using the flow-through technique. Thisguide will probably be useful for conducting early life-stagetoxicity tests with s
4、ome other species of fish, although modi-fications 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 following prescribedprocedures, results of tests conducted using unus
5、ual 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 procedures for conducting earlylife-stage toxicity tests with fishes.1.3 T
6、hese procedures are applicable to all chemicals, eitherindividually or in formulations, commercial products, orknown mixtures, that can be measured accurately at thenecessary concentrations in water. With appropriate modifica-tions these procedures can be used to conduct tests ontemperature, dissolv
7、ed oxygen, and pH and on such materialsas aqueous effluents (see Guide E 1192), leachates, oils,particulate matter, sediments, and surface waters.1.4 This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Standard 4Significance and Use 5.1Hazards 6Apparatus 7Facilitie
8、s 7.1Construction Materials 7.2Metering System 7.3Test Chambers and Incubation Cups 7.4Cleaning 7.5Acceptability 7.6Dilution Water 8Requirements 8.1Source 8.2Treatment 8.3Characterization 8.4Test Material 9General 9.1Stock Solution 9.2Test Concentration(s) 9.3Test Organisms 10Species 10.1Age 10.2Sou
9、rce 10.3Brood Stock 10.4Handling 10.5Procedure 11Experimental Design 11.1Dissolved Oxygen 11.2Temperature 11.3Beginning the Test 11.4Thinning 11.5Feeding 11.6Duration of Test 11.7Biological Data 11.8Other Measurements 11.9Analytical Methodology 12Acceptability of Test 13Calculation of Results 14Docu
10、mentation 15AppendixesAppendix X1 Salmon, Trout, and CharAppendix X2 Northern pikeAppendix X3 Fathead minnowAppendix X4 White suckerAppendix X5 Channel catfishAppendix X6 BluegillAppendix X7 Gulf toadfishAppendix X8 Sheepshead minnowAppendix X9 SilversidesAppendix X10 Statistical GuidanceAppendix X1
11、1. Striped Bass1.5 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is the1This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fate and is the direct responsibility of SubcommitteeE47.01 on Aquatic
12、Assessment and Toxicology.Current edition approved Aug. 1, 2005. Published September 2005. Originallyapproved in 1988. Last previous edition approved in 2004 as E 1241 98(2004).1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.responsi
13、bility of the user 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 6 and 9.2. Referenced Documents2.1 ASTM Standards:2E 380 Practice for Use of the Internati
14、onal System of Units(SI) (the Modernized Metric System)3E 729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE 943 Terminology Relating to Biological Effects and En-vironmental FateE 1023 Guide for Assessing the Hazard of a Material toAquat
15、ic Organisms and Their UsesE 1192 Guide for Conducting Acute Toxicity Tests onAqueous Ambient Samples and Effluents with Fishes,Macroinvertebrates, and AmphibiansE 1203 Practice for Using Brine Shrimp Nauplii as Food forTest Animals in Aquatic Toxicology3. Terminology3.1 The words “must,” “should,”
16、“may,” “can,” and “might”have very specific meanings in this standard. “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
17、 with factors that directly relate 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 resultsquestion
18、able. Terms such as “is desirable,” “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
19、“may” and “can” is preserved, and “might” is never used as asynonym for either “may” or “can.”3.2 For definitions of other terms used in this standard, referto Guide E 729, Terminology E 943, and Guide E 1023. For anexplanation of units and symbols, refer to Practice E 380.3.2.1 antagonisma situatio
20、n which an effect of an expo-sure to multiple substances is less than would be expected if theknown effects of the individual substances were added to-gether.3.2.2 synergisma situation in which an effect of an expo-sure to multiple substances is more than would be expected ifthe known effects of the
21、 individual substances were addedtogether.3.2.3 confoundinga situation in which one or more othervariables covary with the independent variable, making itimpossible to determine the influence of the independentvariable on the dependent variable.4. Summary of Guide4.1 In each of two or more treatment
22、s, embryos and thesubsequent larvae of one species of fish are maintained in twoor more test chambers in a flow-through system for 28 to 120days, depending upon species. In each of the one or morecontrol treatments, the embryos and larvae are maintained indilution water to which no test material has
23、 been added in orderto provide (a) a measure of the acceptability of the test bygiving an indication of the quality of the embryos and larvaeand the suitability of the dilution water, food, test conditions,handling procedures, and so forth, and (b) the basis forinterpreting data obtained from the ot
24、her treatments. In each ofthe one or more other treatments, the embryos and larvae aremaintained in dilution water to which a selected concentrationof test material has been added. Specified data on the concen-tration of test material and the survival and growth of theembryos and larvae in each test
25、 chamber are obtained andanalyzed to determine the effect(s) of the test material on thesurvival and growth of the test organisms.5. Significance and Use5.1 Protection of a species requires prevention of unaccept-able effects on the number, weight, health, and uses of theindividuals of that species.
26、 An early life-stage toxicity testprovides information about the chronic toxicity of a testmaterial to a species of fish. The primary adverse effectsstudied are reduced survival and growth.5.2 Results of early life-stage toxicity tests are generallyuseful estimates of the results of comparable life-
27、cycle testswith the same species (1).4However, results of early life-stagetests are sometimes under estimative of those obtained with thesame species in the longer life-cycle tests (2).5.3 Results of early life-stage toxicity tests might be used topredict long-term effects likely to occur on fish in
28、 fieldsituations as a result of an exposure under comparable condi-tions, except that motile organisms might avoid exposure whenpossible.5.4 Results of early life-stage toxicity tests might be used tocompare the chronic sensitivities of different fish species andthe chronic toxicities of different m
29、aterials, and to study theeffects of various environmental factors on results of such tests.5.5 Results of early life-stage toxicity tests might be animportant consideration when assessing the hazards of materi-als to aquatic organisms (see Guide E 1023) or when derivingwater quality criteria for aq
30、uatic organisms (3).5.6 Results of an early life-stage test might be useful forpredicting the results of chronic tests on the same test materialwith the same species in another water or with another speciesin the same or a different water. Most such predictions take intoaccount the results of acute
31、toxicity tests, and so the usefulnessof the results of an early life-stage test is greatly increased by2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards
32、 Document Summary page onthe ASTM website.3Withdrawn.4Boldface numbers in parentheses refer to the list of references at the end of thisguide.E 1241 052reporting also the results of an acute toxicity test (see GuideE 729) conducted with juveniles of the same species under thesame conditions.5.7 Resu
33、lts of early life-stage toxicity tests might be usefulfor studying the biological availability of, and structure-activity relationships between, test materials.5.8 Results of early life-stage toxicity tests will depend ontemperature, composition of the dilution water, condition of thetest organisms,
34、 and other factors.6. Hazards6.1 Many materials can affect humans adversely if precau-tions are inadequate. Therefore, skin contact with all testmaterials and solutions of them should be minimized by suchmeans as wearing appropriate protective gloves (especiallywhen washing equipment or putting hand
35、s in test solutions),laboratory coats, aprons, and glasses, and by using dip nets,forceps, or tubes, to remove organisms from test solutions.Special precautions, such as covering test chambers andventilating the area surrounding the chambers, should be takenwhen conducting tests on volatile material
36、s. Information ontoxicity to humans (4), recommended handling procedures (5),and chemical and physical properties of the test material shouldbe studied before a test is begun. (WarningSpecial proce-dures might be necessary with radiolabeled test materials (6)and with test materials that are, or are
37、suspected of being,carcinogenic (7).6.2 Although disposal of stock solutions, test solutions, andtest organisms poses no special problems in most cases, healthand safety precautions and applicable regulations should beconsidered before beginning a test. Removal or degradation oftest material might b
38、e desirable before disposal of stock andtest solutions.6.3 Cleaning of equipment with a volatile solvent such asacetone should be performed only in a well-ventilated area inwhich no smoking is allowed and no open flame, such as a pilotlight, is present. (WarningAn acidic solution should not bemixed
39、with a hypochlorite solution because hazardous fumesmight be produced.6.4 To prepare dilute acid solutions, concentrated acidshould be added to water, not vice versa. Opening a bottle ofconcentrated acid and adding concentrated acid to water shouldbe performed only in a fume hood.6.5 Because dilutio
40、n water and test solutions are usuallygood conductors of electricity, use of ground fault systems andleak detectors should be considered to help avoid electricalshocks. Salt water is such a good conductor that protectivedevices are strongly recommended.7. Apparatus7.1 FacilitiesFlow-through tanks sh
41、ould be available forculturing brood stock, and for holding and acclimating testorganisms. The test chambers should be in a constant-temperature area or recirculating water bath. An elevatedheadbox might be desirable so dilution water can be gravity-fed into holding, acclimation, and culture tanks,
42、and themetering system (see 7.3). Strainers and air traps should beincluded in the water-supply system. Headboxes and holding,acclimation, culture, and dilution-water tanks should beequipped for temperature control and aeration (see 8.3). Airused for aeration should be free of fumes, oil, and water;
43、 filtersto remove oil and water are desirable. Filtration of air througha 0.22 m bacterial filter might be desirable.The facility shouldbe well ventilated and free of fumes. To further reduce thepossibility of contamination by test materials and other sub-stances (especially volatile ones) holding,
44、acclimation, andculture tanks should not be in a room in which toxicity tests areconducted, stock solutions or test solutions are prepared, orequipment is cleaned. During holding, acclimation, culture,and testing, organisms should be shielded from disturbanceswith curtains or partitions to prevent u
45、nnecessary stress. Atiming device should be used to control the photoperiod (seeAppendix X1-Appendix X9).A15- to 30-min transition period(8) might be desirable whenever the lights go on to reduce thepossibility of organisms being stressed by large, suddenincreases in light intensity. A transition pe
46、riod when the lightsgo off might also be desirable.7.2 Construction MaterialsEquipment and facilities thatcome in contact with stock solutions, test solutions, or anywater into which test organisms will be placed should notcontain substances that can be leached or dissolved by aqueoussolutions in am
47、ounts that adversely affect fish. In addition,equipment and facilities that contact stock solutions or testsolutions should be chosen to minimize sorption of testmaterials from water.7.2.1 Glass, Type 316 Stainless Steel, Nylon, and Fluoro-carbon PlasticsUse whenever possible to minimize dissolu-tio
48、n, leaching, and sorption, except that stainless steel shouldnot be used for tests on metals in salt water.7.2.2 Concrete and Rigid PlasticsMay be used for hold-ing, acclimation, and culture tanks and in the water-supplysystem, but these materials should be soaked, preferably inflowing dilution wate
49、r, for a week or more before use (9). Castiron pipe should not be used with salt water and probablyshould not be used in a freshwater-supply system becausecolloidal iron will be added to the dilution water and strainerswill be needed to remove rust particles. A specially designedsystem is usually necessary to obtain salt water from a naturalwater source (see Guide E 729). Dilution water, stock solu-tions, or test solutions should not contact brass, copper, lead,galvanized metal, and natural rubber before or during the test.Items made of neoprene rubber or other materials
