ASTM_E_2591_-_07_2013.pdf

Designation:E259107(Reapproved 2013)Standard Guide forConducting Whole Sediment Toxicity Tests withAmphibians1This standard is issued under the fixed designation E2591;the number immediately following the designation indicates the year 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 standard covers procedures for obtaining labora-tory data concerning the toxicity of test material(for example,sediment or hydric soil(that is,a soil that is saturated,flooded,or ponded long enough during the growing season to developanaerobic(oxygen-lacking)conditions that favor the growthand regeneration of hydrophytic vegetation)to amphibians.This test procedure uses larvae of the northern leopard frog(Rana pipiens).Other anuran species(for example,the greenfrog(Rana clamitans),the wood frog(Rana sylvatica),theAmerican toad(Bufo americanus)may be used if sufficientdata on handling,feeding,and sensitivity are available.Testmaterial may be sediments or hydric soil collected from thefield or spiked with compounds in the laboratory.1.2 The test procedure describes a 10-d whole sedimenttoxicity test with an assessment of mortality and selectedsublethal endpoints(that is,body width,body length).Thetoxicity tests are conducted in 300 to 500-mL chamberscontaining 100 mLof sediment and 175 mLof overlying water.Overlying water is renewed daily and larval amphibians are fedduring the toxicity test once they reach Gosner stage 25(operculum closure over gills).The test procedure is designedto assess freshwater sediments,however,R.pipiens can toler-ate mildly saline water(not exceeding about 2500 mg Cl-/L,equivalent to a salinity of about 4.1 when Na+is the cation)in10-d tests,although such tests should always include a con-current freshwater control.Alternative test durations and sub-lethal endpoints may be considered based on site-specificneeds.Statistical evaluations are conducted to determinewhether test materials are significantly more toxic than thelaboratory control sediment or a field-collected referencesample(s).1.3 Where appropriate,this standard has been designed tobe consistent with previously developed methods for assessingsediment toxicity to invertebrates(for example,Hyalella az-teca and Chironomus dilutus toxicity tests)described in theUnited States Environmental ProtectionAgency(USEPA,(1)2freshwater sediment testing guidance,Test Methods E1367 andE1706,and Guides E1391,E1525,E1611,and E1688.Testsextending to 10 d or beyond,and including sublethal measure-ments such as growth,are considered more effective inidentifying chronic toxicity and thus delineating areas ofmoderate contamination(1-3).1.4 Many historical amphibian studies,both water andsediment exposure,have used tests of shorter duration(5 daysor less)(for example,4-7)and,although both survival andsublethal endpoints were often assessed,there is substantiveevidence that tests of longer duration are likely to be moresensitive to some contaminants(8,9).Research performed todevelop and validate this test protocol included long-term(through metamorphosis)investigations and other researchershave also conducted long-duration tests with anurans(7-11).Inthe development of these procedures,an attempt was made tobalance the needs of a practical assessment with the importanceof assessing longer-term effects so that the results will demon-strate the needed accuracy and precision.The most recentsediment toxicity testing protocols for invertebrates haveencompassed longer duration studies which allow the measure-ment of reproductive endpoints(1,12).Such tests,because ofincreased sensitivity of the sublethal endpoints,may also behelpful in evaluating toxicity.Full life-cycle studies withanurans(including reproduction)are usually not feasible fromeither a technical or monetary standpoint.However,if site-specific information indicates that the contaminants present arelikely to affect other endpoints(including teratogenicity),thenthe duration of the toxicity test may be increased throughmetamorphosis or additional sublethal endpoints may be mea-sured(for example,impaired behavior,deformities,time-to-metamorphosis).The possible inclusion of these endpoints andextension of test length should be considered during develop-ment of the project or study plan(see 8.1.1).1.5 The methodology presented in this standard was devel-oped under a Department of Defense(DoD)research programand presented in a guidance manual for risk assessment staff1This 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 March 1,2013.Published March 2013.Originallyapproved in 2007.Last previous edition approved in 2007 as E259107.DOI:10.15