ASTM_D_5660_-_96_2004.pdf

Designation:D 5660 96(Reapproved 2004)Standard Test Method forAssessing the Microbial Detoxification of ChemicallyContaminated Water and Soil Using a Toxicity Test with aLuminescent Marine Bacterium1This standard is issued under the fixed designation D 5660;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(e)indicates an editorial change since the last revision or reapproval.1.Scope1.1 This test method(1)2covers a procedure for the rapidevaluation of the toxicity3of wastewaters and aqueous extractsfrom contaminated soils and sediments,to the luminescentmarine bacterium Photobacterium phosphoreum,4prior to andfollowing biological treatment.This test method is meant foruse as a means to assess samples resulting from biotreatabilitystudies.Sensitivity data for P.phosphoreum to over 1300chemicals have been reported in the literature(2).Some of thepublications are very relevant to this test method(3).The dataobtained from this test method,when combined with respirom-etry,total organic carbon(TOC),biochemical oxygen demand(BOD),chemical oxygen demand(COD),or spectrophotomet-ric data,can assist in the determination of the degree ofbiodegradability of a contaminant in water,soil,or sediment(3).The percentage difference between the IC20 of treated anduntreated sample is used to assess the progress of detoxifica-tion.1.2 This test method is applicable to the evaluation of thetoxicity(to a specific microbe)and its implication on thebiodegradation of aqueous samples from laboratory researchbio-reactors(liquid or soil),pilot-plant biological treatmentsystems,full-scale biological treatment systems,and landapplication processes(see Notes 1 and 2).NOTE1If the biologically treated material is to be discharged in sucha manner as to potentially impact surface waters and ground water,orboth,then the user must consult appropriate regulatory guidance docu-ments to determine the proper test species for evaluating potentialenvironmental impact(4).Correlations between data concerning reductionin toxicity produced by this test method and by procedures for acute orshort-term chronic toxicity tests,or both,utilizing invertebrates and fish(see Guides E 729 and E 1192),should be established,wherever possible.NOTE2Color(especially red and brown),turbidity,and suspendedsolids interfere with this test method by absorbing or reflecting light.Inthese situations data are corrected for these effects by use of an absorbancecorrection procedure included in this test method(see 5.3,6.1,and 6.2).51.3 The results of this test method are reported in terms ofan inhibitory concentration(IC),which is the calculatedconcentration of sample required to produce a specific quanti-tative and qualitative inhibition.The inhibition measured is thequantitative reduction in light output of luminescent marinebacteria(that is,IC20 represents the calculated concentrationof sample that would produce a 20%reduction in the lightoutput of exposed bacteria over a specified time).1.4 The values stated in SI units are to be regarded as thestandard.The values given in parentheses are for informationonly.1.5 This standard does not purport to address 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 hazardstatements are given in Section 9.2.Referenced Documents2.1 ASTM Standards:6D 888 Test Methods for Dissolved Oxygen in Water1This test method is under the jurisdiction of ASTM Committee D34 on WasteManagement and is the direct responsibility of Subcommittee D34.07 on MunicipalSolid Waste.Current edition approved March 10,1996.Published May 1996.Originallypublished as D 5660 95.Last previous edition D 5660 95.2The boldface numbers in parentheses refer to the list of references at the end ofthis standard.3Toxicity measured as toxic inhibition of bacterial light output.4Microbics Corp.is currently the only known supplier of the reagents(testorganism Photobacterium phosphoreum strain NRRL B-11177)specific to this testmethod.There are two known manufacturers of analyzers that can be used tomeasure bioluminescence under temperature control:Microbics Corp.,2232 Ruth-erford Road,Carlsbad,CA 92008(Microtox Model 500 and Model 2055 Analyz-ers),and Pharmacia LKB,9319 Gaither Road,Gaithersburg,MD 20877(LKBWallac Model 1250 and Model 1251 Luminometers).Other instruments would beconsidered when they become available.Please notifyASTM Subcommittee D34.09if you are aware of any additional systems or instruments capable of performing thistesting.5At present(1993)use of the color correction scheme described in thisprocedure is known to be effective only with the Microbics Corporations toxicityanalyzers,due to the fact that the co