1、Designation: E 1562 00 (Reapproved 2006)Standard Guide forConducting Acute, Chronic, and Life-Cycle Aquatic ToxicityTests with Polychaetous Annelids1This standard is issued under the fixed designation E 1562; the number immediately following the designation indicates the year oforiginal adoption or,
2、 in the case of revision, the year of last 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 data con-cerning the adverse effects
3、 of a test material added to marineand estuarine waters on certain species of polychaetes duringshort- or long-term continuous exposure. The polychaetespecies used in these tests are taken from laboratory culturesand exposed to varying concentrations of a toxicant in staticconditions. These procedur
4、es may be useful for conductingtoxicity tests with other species of polychaetes, althoughmodifications might be necessary.1.2 Modifications of these procedures might be justified byspecial needs or circumstances. Although using appropriateprocedures is more important than following prescribed pro-ce
5、dures, the results of tests conducted using unusual proce-dures are not likely to be comparable to those of many othertests. Comparisons of results obtained using modified andunmodified versions of these procedures might provide usefulinformation concerning new concepts and procedures forconducting
6、acute, chronic, or life-cycle tests with other speciesof polychaetes.1.3 These procedures are applicable to most chemicals,either individually or in formulations, commercial products,and known or unknown mixtures. With appropriate modifica-tions, these procedures can be used to conduct these tests o
7、nfactors such as temperature, salinity, and dissolved oxygen.These procedures can also be used to assess the toxicity ofpotentially toxic discharges such as municipal wastes, oildrilling fluids, produced water from oil well production, andother types of industrial wastes. An LC50 (medial lethalconce
8、ntration) is calculated from the data generated in eachacute and chronic toxicity test. Reproductive success and thenumber of offspring produced are used to measure the effect ofa toxicant on life-cycle tests; data are analyzed statistically toindicate that concentration at which a significant diffe
9、renceoccurs between the test solutions and control(s).1.4 The results of acute or chronic toxicity tests withtoxicants added experimentally to salt water should usually bereported in terms of an LC50. The results of life-cycle toxicitytests with toxicants added experimentally to salt water shouldbe
10、reported as that concentration at which a statisticallysignificant difference in the number of offspring is producedwith reference to the control(s).1.5 This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Apparatus 6Facilities 6.1Constr
11、uction Materials 6.2Test Chambers 6.3Cleaning 6.4Acceptability 6.5Safety Precautions 7Dilution Water 8Requirements 8.1Source 8.2Treatment 8.3Characterization 8.4Test Material 9General 9.1Stock Solution 9.2Test Concentrations 9.3Test Organisms 10Species 10.1Age 10.2Source 10.3Feeding 10.4Holding 10.5
12、Quality 10.6Procedure 11Experimental Design 11.1Acute Test 11.1.1Chronic Test 11.1.2Life-Cycle Test 11.1.3Test Condition Specifications 11.2Dissolved Oxygen 11.2.1Temperature 11.2.2Loading 11.2.3Salinity 11.2.4Light 11.2.5Beginning the Test 11.3Feeding 11.4Duration of Test 11.5Biological Data 11.6Ot
13、her Measurements 11.71This 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 approved April 1, 2006. Published May 2006. Originallyapproved in 1
14、994. Last previous edition approved in 2000 as E 1562 00.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.Analytical Methodology 12Acceptability of Test 13Calculation of Results 14Report 15Keywords 16Appendixes:Neanthes arenaceodentat
15、a Appendix X1Capitella capitata Appendix X2Ophryotrocha diadema Appendix X3Dinophilus gyrociliatus Appendix X41.6 The values stated in SI units are to be regarded as thestandard.1.7 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is therespons
16、ibility 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 precau-tionary statements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:2E 729 Guide for Conducting Acute To
17、xicity 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 UsesIEEE/ASTM SI 10 American National Standard for Use ofthe Interna
18、tional System of Units (SI): The Modern MetricSystem3. Terminology3.1 Definitions:3.1.1 The words “must,” “should,” “may,” “can,” and“might” have very specific meanings in this guide. “Must” isused to express the strongest possible recommendation, justshort of an absolute requirement, that is, to st
19、ate that this testought to be designed to satisfy the specific condition, unless thepurpose of the test requires a different design. “Must” is usedonly in connection with factors that relate directly to theacceptability of the test (see Section 13). “Should” is used tostate that the specific conditi
20、on is recommended and ought tobe met, if possible.Although violation of one“ should” is rarelya serious matter, violation of several will often render theresults questionable. Terms such as “is desirable,” “is oftendesirable,” and “might be desirable” are used in connectionwith less important factor
21、s. “May” is used to mean “is (are)allowed to,” “can” is used to mean “is (are) able to,” and“might” is used to mean “could possibly.” Thus the classicdistinction between “may” and “can” is preserved, and “might”is never used as a synonym for either “may” or “can.”3.1.2 For definitions of other terms
22、 used in this guide, referto Guide E 729, Terminology E 943, and Guide E 1023. For anexplanation of units and symbols, refer to IEEE/ASTM SI 10.4. Summary of Guide4.1 The toxicity of a substance in marine or estuarine waterscan be determined through a 96-h acute or chronic toxicity test(see 11.1.2)
23、in 100-mm diameter glass petri dishes for Nean-thes arenaceodentata and Capitella capitata. Stender dishesare recommended for acute, chronic, and life-cycle tests withthe smaller species Ophryotrocha diadema and Dinophilusgyrociliatus. Petri dishes provide ample horizontal space tominimize cannibali
24、sm in the case of Neanthes arenaceoden-tata. Larger containers, such as gallon jars, are necessary forlife-cycle tests using Neanthes arenaceodentata and Capitellacapitata. The static-renewal technique is recommended for thechronic and life-cycle tests with these latter species; it isusually not nec
25、essary to renew the solutions in 96-h tests. Witheither the static or static-renewal system, data on the concen-tration of test material are obtained and analyzed to determinethe effect(s) of the toxicant on survival and reproduction.5. Significance and Use5.1 Polychaetes are an important component
26、of the benthiccommunity, in which they generally comprise 30 to 50 % of themacroinvertebrate population. They are preyed upon by manyspecies of fish, birds, and larger invertebrate species. Largerpolychaetes feed on small invertebrates, larval stages ofinvertebrates, and algae. Polychaetes are espec
27、ially sensitive toinorganic toxicants and, to a lesser extent, to organic toxicants(1).3The ecological importance of polychaetes and their widegeographical distribution, ability to be cultured in the labora-tory, and sensitivity to contaminants make them appropriateacute and chronic toxicity test or
28、ganisms. Their short life cycleenables the investigator to measure the effect of contaminantson reproduction.5.2 An acute toxicity or chronic text is conducted to obtaininformation concerning the immediate effects of an exposure toa test material on a test organism under specified experimentalcondit
29、ions. An acute toxicity test provides data on the short-term effects, which are useful for comparisons to other speciesbut do not provide information on delayed effects. Chronictoxicity tests provide data on long-term effects.5.3 A life-cycle toxicity test is conducted to determine theeffects of the
30、 test material on survival, growth, and reproduc-tion of the test species. Additional sublethal endpoints (forexample, biochemical, physiological, and histopathological)may be used to determine the health of the species under fieldconditions.5.4 The results of acute, chronic, and life-cycle toxicity
31、 testscan be used to predict effects likely to occur on marineorganisms under field conditions.5.5 The results of acute, chronic, or life-cycle toxicity testsmight be used to compare the sensitivities of different speciesand the toxicities of different test materials, as well as to studythe effects
32、of various environmental factors on the results ofsuch tests.5.6 The results of acute, chronic, or life-cycle toxicity testsmight be an important consideration when assessing the2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org.
33、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 1562 00 (2006)2hazards of materials to marine organisms (see Guide E 1023)or when derivi
34、ng water quality criteria for aquatic organisms(2).5.7 The results of acute, chronic, or life-cycle toxicity testsmight be useful for studying the biological availability of, andstructure activity relationships between, test materials.5.8 The results of acute, chronic, and life-cycle toxicity testsw
35、ill depend partly on the temperature, quality of food, condi-tion of test organisms, test procedures, and other factors.6. Apparatus6.1 FacilitiesAquaria (4 to 57 L) or gallon jars containingeither clean (uncontaminated), natural, or reconstituted seawater should be used for culturing these four spe
36、cies ofpolychaetes or holding them after field collection and prior toa test. Aquaria provided with aeration have been used success-fully for culturing and holding polychaetes. The holding tanksand any area used for culturing or holding polychaetes shouldbe located in a room or space separated from
37、that in whichtoxicity tests are to be conducted. The sea water should beanalyzed periodically to ensure a constant salinity. The holdingaquaria, water supply, or room in which they are kept should beequipped with temperature control.Aeration can be provided toensure that dissolved oxygen is greater
38、than 60 % saturationand that water circulation is adequate in the aquaria. Thesespecies of polychaetes do not require a definite light regime.6.2 Construction MaterialsEquipment and facilities thatcontact stock solutions, test solutions, or any water into whichtest organisms will be placed should no
39、t contain substancesthat can be leached or dissolved by aqueous solutions inamounts that affect the test organisms adversely. In addition,equipment and facilities that contact stock or test solutionsshould be chosen to minimize the sorption of test materialsfrom water. Glass, Type 316 stainless stee
40、l, nylon, high-densitypolyethylene, polycarbonate, and fluorocarbon plastics shouldbe used whenever possible to minimize dissolutions, leaching,and sorption, except that stainless steel should not be used intests on metals in salt water. Concrete and rigid plastics may beused for holding tanks and i
41、n the water-supply system, but theyshould be soaked, preferably in flowing dilution water, for oneweek or more before use (3). Cast iron pipe should not be usedwith salt water and probably should not be used in a freshwater-supply filter system because colloidal iron will be addedto the dilution wat
42、er and strainers will be necessary to removerust particles. A specially designed system is usually necessaryto obtain salt water from a natural water source (4). Brass,copper, lead, galvanized metal, and natural rubber should notcontact the dilution water, stock solutions, or test solutionsbefore or
43、 during the test. Items made of neoprene rubber orother materials not mentioned previously should not be usedunless it has been shown that their use will not affect either thesurvival, growth, or reproduction of polychaetes adversely.6.3 Test ChambersIn a toxicity test with marine organ-isms, test c
44、hambers are defined as the smallest physical unitsbetween which there are no water connections. Glass petridishes measuring 100 mm in diameter and 20 mm in height arethe most satisfactory chambers for use in acute and chronictoxicity tests with Neanthes arenaceodentata and Capitellacapitata. Only on
45、e Neanthes arenaceodentata should be placedin a single chamber since this species can be cannibalistic.Stender dishes measuring 40 mm in diameter and providedwith a ground glass cover are the most satisfactory chambersfor acute, chronic, and life-cycle tests with Ophryotrochadiadema and Dinophilus g
46、yrociliatus. Capitella capitata,Ophryotrocha diadema, and Dinophilus gyrociliatus are notcannibalistic, and 5 to 10 specimens may be placed within asingle chamber. For reproductive tests, glass chambers mea-suring 3.79 L and containing 2.5 L of test solution should beused with N. arenaceodentata and
47、 C. capitata. Aeration mustbe provided. Chambers the same size as that used in the acuteand chronic tests with O. diadema and D. gyrociliatus can beused for the reproductive tests. The chambers should becovered to keep out extraneous contaminants and to reduce theevaporation of test solution and tes
48、t material. All chambers ina test must be identical.6.4 CleaningTest chambers and other glassware, andequipment used to store and prepare the test sea water, stocksolutions, and test sediments, should be cleaned before use.New items should be cleaned before each use by washing withlaboratory deterge
49、nt and rinsing with water, a weak-miscibleorganic solvent, and acid (10 % nitric or hydrochloric acid),and they should be rinsed twice with distilled, deionized, ordilution water. Metals, sulfides, and carbonate deposits areremoved by the acid rinse, and organics are removed by theorganic solvent rinse. A dichromate-sulfuric acid cleaningsolution may be used in place of both the organic solvent andacid rinses, but it might attack silicone adhesives.At the end ofeach test, all items that are to be used again should immediatelybe (1) emptied; (2) rinsed with water; (3) cleaned by a
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