1、Designation: E 1193 97 (Reapproved 2004)Standard Guide forConducting Daphnia magna Life-Cycle Toxicity Tests1This standard is issued under the fixed designation E 1193; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of la
2、st revision. A 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 covers procedures for obtaining laboratorydata concerning the adverse effects of a test material (added todi
3、lution water, but not to food) on Daphnia magna Straus,1820, during continuous exposure throughout a life-cycle usingthe renewal or flow-through techniques. These procedures alsoshould be useful for conducting life-cycle toxicity tests withother invertebrate species, although modifications might ben
4、ecessary.1.2 These procedures are applicable to most chemicals,either individually or in formulations, commercial products, orknown mixtures. With appropriate modifications, these proce-dures can be used to conduct tests on temperature, dissolvedoxygen, pH, and on such materials as aqueous effluents
5、 (alsosee Guide E 1192), leachates, oils, particulate matter, sedi-ments, and surface waters. The technique, (renewal or flow-through), will be selected based on the chemical characteristicsof the test material such as high oxygen demand, volatility,susceptibility to transformation (biologically or
6、chemically), orsorption to glass.1.3 Modification of these procedures might be justified byspecial needs or circumstances. Although using appropriateprocedures is more important than following prescribed pro-cedures, results of tests conducted using unusual proceduresare not likely to be comparable
7、to results of standard testprocedures. Comparison of results obtained using modified andunmodified versions of these procedures might provide usefulinformation on new concepts and procedures for conductinglife-cycle toxicity tests with D. magna.1.4 This guide is arranged as follows:SectionReferenced
8、 Documents 2Terminology 3Summary of Guide 4Significance and Use 5Apparatus 6Facilities 6.1Construction Materials 6.2SectionTest Chambers 6.3Cleaning 6.4Acceptability 6.5Reagents 7Purity of Reagents 7.1Hazards 8Dilution Water 9Requirements 9.1Source 9.2Treatment 9.3Characterization 9.4Test Material 1
9、0General 10.1Stock Solutions 10.2Test Concentrations(s) 10.3Test Organisms 11Species 11.1Age 11.2Source 11.3Brood Stock 11.4Food 11.5Handling 11.6Harvesting Young 11.7Quality 11.8Procedure 12Experimental Design 12.1Dissolved Oxygen 12.2Temperature 12.3Loading 12.4Selection of Test System 12.5Beginni
10、ng the Test 12.6Care and Maintenance 12.7Feeding 12.8Duration 12.9Biological Data 12.10Other Measurements 12.11Analytical Methodology 13Acceptability of Test 14Calculation of Results 15Report 16Keywords 17AppendixesAppendix X1 Statistical GuidanceAppendix X2 Food1.5 The values stated in SI units are
11、 to be regarded as thestandard. The values given in parentheses are for informationonly.1This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fate and is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition
12、approved April 1, 2004. Published April 2004. Originallyapproved in 1987. Last previous edition approved in 1997 as E 1193 97.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.1.6 This standard does not purport to address all of thesaf
13、ety concerns, if any, associated with its use. It is theresponsibility 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 8.2. Referenced Documents2
14、.1 ASTM Standards:2E 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 toAquatic Organisms and Their UsesE 1192 Gui
15、de for Conducting Acute Toxicity Tests onAqueous Ambient Samples and Effluents with Fishes,Macroinvertebrates, and AmphibiansIEEE/ASTM-SI-10 Standard for Use of the InternationalSystem of Units (SI):The Modern Metric System3. Terminology3.1 The words “must,” “should,”“ may,” “can,” and “might”have v
16、ery specific meanings in this guide.3.2 mustused to express an absolute requirement, that is,to state that the test ought to be designed to satisfy the specifiedcondition, unless the purpose of the test requires a differentdesign. “Must” is used only in connection with factors thatdirectly relate to
17、 the acceptability of the test (see 14.1).3.3 shouldused to state that the specified condition isrecommended and ought to be met if possible. Althoughviolation of one “should” is rarely a serious matter, violation ofseveral will often render the results questionable. Terms suchas “is desirable,” “is
18、 often desirable,” and “might be desirable”are used in connection with less important factors.3.4 mayused to mean “is (are) allowed to,” “can” is usedto mean “is (are) able to,” and “might” is used to mean “couldpossibly.” Therefore the classic distinction between “may” and“can” is preserved, and “m
19、ight” is never used as a synonym foreither“ may” or “can.”3.5 For definitions of other terms used in this guide, refer toGuide E 729 and Terminology E 943. For an explanation ofunits and symbols, refer to IEEE/ASTM-SI-10.4. Summary of Guide4.1 A 21-day life-cycle toxicity test for Daphnia magna isde
20、scribed. The test design allows for the test organisms to beexposed to a toxicant using either the renewal technique (withexchange of the total volume of test water and toxicant at leastthree times a week) or the flow-through technique (withcontinual water and toxicant addition, usually at least fou
21、rvolume additions per day). At least five concentrations of a testmaterial, a control, and a solvent control (if applicable)replicated at least four times are recommended. Each testconcentration has at least ten Daphnia per treatment. Thetechnique (renewal or flow-through) which uses a minimum often
22、 daphnids per treatment has only one daphnid per replicate,whereas the typical technique (renewal or flow-through) uti-lizes four replicates with at least five daphnids per replicate($20 daphnids per treatment). A control consists of maintain-ing daphnids in dilution water to which no test material
23、hasbeen added to provide (1) a measure of the acceptability of thetest by giving an indication of the quality of the test organismsand the suitability of the dilution water, food, test conditions,handling procedures, and so forth, and (2) the basis forinterpreting data obtained from the other treatm
24、ents. In each ofthe other treatments, the daphnids are maintained in dilutionwater, to which a selected concentration of test material hasbeen intentionally added. Measurement end points obtainedduring the test include the concentration of the test material andfinal number alive, final weight, and n
25、umber of progeny perdaphnid. Then data are analyzed to determine the effect of thetest material on survival, growth, and reproduction of D.magna.5. Significance and Use5.1 Protection of an aquatic species requires prevention ofunacceptable effects on populations in natural habitats. Toxic-ity tests
26、are conducted to provide data that may be used topredict what changes in numbers and weights of individualsmight result from similar exposure to the test material in thenatural aquatic environment. Information might also be ob-tained on the effects of the material on the health of the species.5.2 Re
27、sults of life-cycle tests with D. magna are used topredict chronic effects likely to occur on daphnids in fieldsituations as a result of exposure under comparable conditions.5.2.1 Life-cycle tests with D. magna are used to comparethe chronic sensitivities of different species, the chronic tox-icitie
28、s of different materials, and study the effects of variousenvironmental factors on the results of such tests.5.2.2 Life-cycle tests with D. magna are used to assess therisk of materials to aquatic organisms (see Guide E 1023) orderive water quality criteria for aquatic organisms (1).35.2.3 Life-cycl
29、e tests with D. magna are used to predict theresults of chronic toxicity tests on the same test material withthe same species in another water or with another species in thesame or a different water. Most such predictions take intoaccount the results of acute toxicity tests, and so the usefulnessof
30、the results of a life-cycle test with D. magna is greatlyincreased by also reporting the results of an acute toxicity test(see Guide E 729) conducted under the same conditions. Inaddition to conducting an acute toxicity test with unfed D.magna, it may be desirable to conduct an acute test in whichth
31、e daphnids are fed the same as in the life-cycle test to see ifthe presence of that concentration of that food affects theresults of the acute test and the acute-chronic ratio (ACR) (see10.3.1).2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at s
32、erviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3The boldface numbers in parentheses refer to the list of references at the end ofthis guide.E 1193 97 (2004)25.2.4 Life-cycle tests are used to evaluate the biologica
33、lavailability of, and structure-activity relationships between,test materials and test organisms.5.3 Results of life-cycle tests with D. magna might beinfluenced by temperature (2), quality of food, composition ofdilution water, condition of test organisms, and other factors.6. Apparatus6.1 Faciliti
34、esCulture and test chambers are often kept ina room maintained at about 20C but at separate locations.Alternatively, culture and test chambers may be placed in atemperature-controlled water bath or environmental chamberor incubator. The water-supply system should provide anadequate supply of dilutio
35、n water to the culture tanks and testchambers. The water-supply system should be equipped fortemperature control and aeration, and strainers and air trapsshould be included in the water-supply system. Air used foraeration should be free of fumes, oil, and water; filters toremove oil and water are de
36、sirable. Filtration of air through a0.22-m bacterial filter might be desirable (3). During culturingand testing, daphnids should be shielded from disturbances toprevent unnecessary stress. The test facility should be well-ventilated and free of fumes. A timing device should be used toprovide a 16-h
37、light and 8-h dark photoperiod (4).A15to30-min transition period when lights go on might be desirableto reduce the possibility of daphnids being stressed by instan-taneous illumination; a transition period when lights go off mayalso be desirable.6.1.1 When D. magna are fed algae, a high-light intens
38、itymight cause sufficient photosynthesis to result in an increase ofpH high enough to kill daphnids (5). Therefore, the maximumacceptable intensity is dependent on the buffer capacity of thedilution water, species, and density of algae, and the kind oftest chamber and cover. Light intensities up to
39、600 lx or afluence rate of 1 w/m2will usually be acceptable, but higherintensities might result in an unacceptably high pH in theculture water.6.2 Construction MaterialsEquipment and facilities thatcontact stock solutions, test solutions, or any water into whichdaphnids will be placed should not con
40、tain substances that canbe leached or dissolved by aqueous solutions in amounts thatcan adversely affect daphnids. In addition, equipment andfacilities that contact stock solutions or test solutions should bechosen to minimize sorption of test materials from water.Glass, Type 316 stainless steel, ny
41、lon, fiberglass, silicon, andfluorocarbon plastics should be used whenever possible tominimize leaching, dissolution, and sorption. Concrete andrigid (unplasticized) plastics may be used for culture tanks andin the water-supply system, but they should be soaked, prefer-ably in flowing dilution water
42、, for several days before use (6).Cast-iron pipe may be used in supply systems, but colloidaliron probably will be added to the dilution water and strainerswill be needed to remove rust particles. Copper, brass, lead,galvanized metal, and natural rubber should not contact dilu-tion water, stock solu
43、tions, or test solutions before or duringthe test. Items made of neoprene rubber and other materials notpreviously mentioned should not be used unless it has beenshown that their use will not adversely affect survival, growth,and reproduction of D. magna (see Section 14).6.3 Test Chambers:6.3.1 Flow
44、-through tests, 500-mL to 2-L glass beakers (orequivalent) with a notch (approximately 4 by 13 cm) cut in thelip may be used to expose the Daphnia to the test material. Thenotch should be covered with 0.33-mm opening (U.S. standardsieve size No. 50) stainless steel or polyethylene screeningsmall eno
45、ugh to retain first instar Daphnia. The screen can beattached to the beaker with silicone adhesive. The chambersshould provide at least 30 mL of solution for each of the initialtest daphnid(s).6.3.2 Renewal tests, beaker ranging in size from 100 to 1000mL. A notched chamber is not required for a ren
46、ewal test. Eachchamber should provide at least 40 mL of solution for each ofthe initial test daphnid(s).6.3.3 Any container made of glass, Type 316 stainless steel,or a fluorocarbon plastic may be used if (1) each chamber isseparate with no interconnections, (2) each chamber contains atleast 30 mL o
47、f test solution (see 12.4) per first-generationdaphnid for flow-through tests and at least 40 mL for renewaltests, (3) there is at least 1000 mm2of air to water interface perdaphnid, and (4) the test solution is at least 30 mm deep. Statictest chambers should be covered with glass, stainless steel,n
48、ylon, or fluorocarbon plastic covers to keep out extraneouscontaminants and to reduce evaporation of test solution. Allchambers and covers in a test must be identical. Covers are notrequired for flow-through studies.6.4 CleaningTest chambers and equipment used to pre-pare and store dilution water, s
49、tock solutions, and test solutionsshould be cleaned before use. New equipment should bewashed with detergent and rinsed with water, a water-miscibleorganic solvent, water, acid (such as 5 % concentrated nitricacid), and washed at least twice with distilled, deionized, ordilution water. Some lots of some organic solvents might leavea film that is insoluble in water. Also, stronger nitric acid, forexample, 10 %, might cause deterioration of silicone adhesive;an initial rinse with 10 % concentrated hydrochloric acid mightprevent such deterioration. A dichromate-sulfuric acid cle
