1、Designation: E1562 00 (Reapproved 2013)Standard Guide forConducting Acute, Chronic, and Life-Cycle Aquatic ToxicityTests with Polychaetous Annelids1This standard is issued under the fixed designation E1562; the number immediately following the designation indicates the year oforiginal adoption or, i
2、n 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 data con-cerning the adverse effects of
3、 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 procedures
4、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 prescribedprocedures
5、, the results of tests conducted using unusual pro-cedures 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 acute
6、, 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 appropriatemodifications, these procedures can be used to conduct thesetests on factor
7、s such as temperature, salinity, and dissolvedoxygen. These procedures can also be used to assess thetoxicity of potentially toxic discharges such as municipalwastes, oil drilling fluids, produced water from oil wellproduction, and other types of industrial wastes. An LC50(medial lethal concentratio
8、n) is calculated from the datagenerated in each acute and chronic toxicity test. Reproductivesuccess and the number of offspring produced are used tomeasure the effect of a toxicant on life-cycle tests; data areanalyzed statistically to indicate that concentration at which asignificant difference oc
9、curs between the test solutions andcontrol(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 reported
10、 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 6.1Construction M
11、aterials 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 10.5Quality
12、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 11.6Other Meas
13、urements 11.7Analytical Methodology 121This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fateand is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved March 1, 2013. Published March 2013. Origi
14、nallyapproved in 1994. Last previous edition approved in 2006 as E1562 00(2006).DOI: 10.1520/E1562-00R13.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1Acceptability of Test 13Calculation of Results 14Report 15Keywords 16Appendixes:N
15、eanthes arenaceodentata Appendix X1Capitella capitata Appendix X2Ophryotrocha diadema Appendix X3Dinophilus gyrociliatus Appendix X41.6 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.1.7 This standard does not purport to addres
16、s all of thesafety 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 precau-tionary statements are given in Section 7.
17、2. Referenced Documents2.1 ASTM Standards:2E729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE943 Terminology Relating to Biological Effects and Envi-ronmental FateE1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and
18、 Their UsesIEEE/ASTM SI 10 American National Standard for Use ofthe International 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 po
19、ssible recommendation, justshort of an absolute requirement, that is, to state 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 th
20、e test (see Section 13). “Should” is used tostate that the specific condition 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,
21、” and “might be desirable” are used in connectionwith less important factors. “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
22、as a synonym for either “may” or “can.”3.1.2 For definitions of other terms used in this guide, referto Guide E729, Terminology E943, and Guide E1023. 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
23、be determined through a 96-h acute or chronic toxicity test(see 11.1.2) 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 Dinophilusgyroci
24、liatus. Petri dishes provide ample horizontal space tominimize cannibalism 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 thechro
25、nic and life-cycle tests with these latter species; it isusually not necessary 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 reprodu
26、ction.5. Significance and Use5.1 Polychaetes are an important component 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 invert
27、ebrates, larval stages ofinvertebrates, and algae. Polychaetes are especially 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 thelaboratory, and sensitivity t
28、o contaminants make them appro-priate acute and chronic toxicity test organisms. Their short lifecycle enables the investigator to measure the effect of contami-nants on reproduction.5.2 An acute toxicity or chronic text is conducted to obtaininformation concerning the immediate effects of an exposu
29、re toa test material on a test organism under specified experimentalconditions. 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.
30、5.3 A life-cycle toxicity test is conducted to determine theeffects of the 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
31、fieldconditions.5.4 The results of acute, chronic, and life-cycle toxicity 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 t
32、he toxicities of different test materials, as well as to studythe effects of various environmental factors on the results ofsuch tests.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume inf
33、ormation, 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.E1562 00 (2013)25.6 The results of acute, chronic, or life-cycle toxicity testsmight be an important consideration when assessing the
34、hazards of materials to marine organisms (see Guide E1023)orwhen deriving 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 mat
35、erials.5.8 The results of acute, chronic, and life-cycle toxicity testswill 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), natura
36、l, or reconstituted seawater should be used for culturing these four species 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
37、 holding polychaetes shouldbe located in a room or space separated from 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 con
38、trol.Aeration can be provided toensure that dissolved oxygen is greater 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 s
39、olutions, or any water into whichtest organisms will be placed should not 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
40、the sorption of test materialsfrom water. Glass, Type 316 stainless steel, 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 sa
41、lt water. Concrete and rigid plastics may beused for holding tanks and in 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-supp
42、ly filter system because colloidal iron will be addedto the dilution water 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 no
43、tcontact the dilution water, stock solutions, or test solutionsbefore or 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 adve
44、rsely.6.3 Test ChambersIn a toxicity test with marineorganisms, test chambers are defined as the smallest physicalunits between which there are no water connections. Glasspetri dishes measuring 100 mm in diameter and 20 mm inheight are the most satisfactory chambers for use in acute andchronic toxic
45、ity tests with Neanthes arenaceodentata andCapitella capitata. Only one Neanthes arenaceodentata shouldbe placed in a single chamber since this species can becannibalistic. Stender dishes measuring 40 mm in diameter andprovided with a ground glass cover are the most satisfactorychambers for acute, c
46、hronic, and life-cycle tests with Oph-ryotrocha diadema and Dinophilus gyrociliatus. Capitellacapitata, Ophryotrocha diadema, and Dinophilus gyrociliatusare not cannibalistic, and 5 to 10 specimens may be placedwithin a single chamber. For reproductive tests, glass chambersmeasuring 3.79 L and conta
47、ining 2.5 L of test solution shouldbe used with N. arenaceodentata and C. capitata. Aerationmust be provided. Chambers the same size as that used in theacute and chronic tests with O. diadema and D. gyrociliatuscan be used for the reproductive tests. The chambers should becovered to keep out extrane
48、ous contaminants and to reduce theevaporation of test solution and test 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 ite
49、ms should be cleaned before each use by washing withlaboratory detergent 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 sho
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