1、Designation: E 2122 02 (Reapproved 2007)Standard Guide forConducting In-situ Field Bioassays With Caged Bivalves1This standard is issued under the fixed designation E 2122; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year o
2、f 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 describes procedures for conducting con-trolled experiments with caged bivalves under field conditions.T
3、he purpose of this approach is to facilitate the simultaneouscollection of field data to help characterize chemical exposureand associated biological effects in the same organism underenvironmentally realistic conditions. This approach of charac-terizing exposure and effects is consistent with the U
4、S EPAecological risk assessment paradigm. Bivalves are useful testorganisms for in-situ field bioassays because they (1) concen-trate and integrate chemicals in their tissues and have a morelimited ability to metabolize most chemicals than other species,(2) exhibit measurable sublethal effects assoc
5、iated with expo-sure to those chemicals, (3) provide paired tissue chemistry andresponse data which can be extrapolated to other species andtrophic levels, (4) provide tissue chemistry data which can beused to estimate chemical exposure from water or sediment,and (5) facilitate controlled experiment
6、ation in the field withlarge sample sizes because they are easy to collect, cage, andmeasure (1, 2)2. The experimental control afforded by thisapproach can be used to place a large number of animals of aknown size distribution in specific areas of concern to quantifyexposure and effects over space a
7、nd time within a clearlydefined exposure period. Chemical exposure can be estimatedby measuring the concentration of chemicals in water, sedi-ment, or bivalve tissues, and effects can be estimated withsurvival, growth, and other sublethal end points (3). Althougha number of assessments have been con
8、ducted using bivalvesto characterize exposure by measuring tissue chemistry orassociated biological effects, relatively few assessments havebeen conducted to characterize both exposure and biologicaleffects simultaneously (2, 4, 5). This guide is specificallydesigned to help minimize the variability
9、 in tissue chemistryand response measurements by using a practical uniform sizerange and compartmentalized cages for multiple measurementson the same individuals.1.2 The test is referred to as a field bioassay because it isconducted in the field and because it includes an element ofrelative chemical
10、 potency to satisfy the bioassay definition.Relative potency is established by comparing tissue concen-trations with effects levels for various chemicals with toxicityand bioaccumulation end points (6, 7, 8, 9, 10) even thoughthere may be more uncertainty associated with effects mea-surements in fie
11、ld studies. Various pathways of exposure canbe evaluated because filter-feeding and deposit-feeding are theprimary feeding strategies for bivalves. Filter-feeding bivalvesmay be best suited to evaluate the bioavailability and associ-ated effects of chemicals in the water column (that is, dissolvedan
12、d suspended particulates); deposit-feeding bivalves may bebest suited to evaluate chemicals associated with sediments(11, 12). It may be difficult to demonstrate pathways ofexposure under field conditions, particularly since filter-feeding bivalves can ingest suspended sediment and facultativedeposi
13、t-feeding bivalves can switch between filter- and depositfeeding over relatively small temporal scales. Filter-feedingbivalves caged within 1 m of bottom sediment have also beenused effectively in sediment assessments from depths of 10 to650 m (5, 13, 14). Caged bivalve studies have also beenconduct
14、ed in the intertidal zone (15). The field testing proce-dures described here are useful for testing most bivalvesalthough modifications may be necessary for a particularspecies.1.3 These field testing procedures with caged bivalves areapplicable to the environmental evaluation of water andsediment i
15、n marine, estuarine, and freshwater environmentswith almost any combination of chemicals, and methods arebeing developed to help interpret the environmental signifi-cance of accumulated chemicals (6, 7, 9, 16, 17). Theseprocedures could be regarded as a guide to an exposure systemto assess chemical
16、bioavailability and toxicity under natural,site- specific conditions, where any clinical measurements arepossible.1.4 Tissue chemistry results from short- and long-termexposures can be reported in terms of concentrations ofchemicals in bivalve tissues (for example, g/g), amount (thatis, weight or ma
17、ss) of chemical per animal (for example,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 approved Oct. 1, 2007. Published October 2007. O
18、riginallyapproved in 2001. Last previous edition approved in 2002 as E 2122 02.2The boldface numbers in parentheses refer to references at the end of thisstandard.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.g/animal), rate of upt
19、ake, or bioaccumulation factor (BAF, theratio between the concentration of a chemical in bivalve tissuesand the concentration in the external environment, includingwater, sediment, and food). Tissue chemistry results can onlybe used to calculate a BAF because caged bivalves in the fieldare exposed t
20、o multiple sources of chemicals and can accumu-late chemicals from water, sediment, and food. Toxicity resultscan be reported in terms of survival (3, 18), growth rate (3, 18),or reproductive effects (19, 20) after a defined exposure period.1.5 Other modifications of these procedures might be justi-
21、fied by special needs or circumstances. Although using appro-priate procedures is more important than following prescribedprocedures, results of tests conducted using unusual proceduresare not likely to be comparable to results of standardized tests.Comparisons of results obtained using modified and
22、 unmodi-fied versions of these procedures might provide useful infor-mation concerning new concepts and procedures for conduct-ing field bioassays with bivalves.1.6 This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Interferences 6Haza
23、rds 7Experimental Design 8Apparatus 9FacilitiesConstruction MaterialsCagesTest Organisms 10SpeciesCommonly Used TaxaSize and Age of Test OrganismsSourceNumber of SpecimensCollectionHandlingHoldingAnimal QualityField Procedures 11Test Initiation: PresortFinal Measurements and DistributionAttachment o
24、f PVC FramesDeploymentRetrieval and End-of-Test MeasurementsAnalysis of Tissues for Background ContaminationCollection and Preparation of Tissues for AnalysisQuality Assurance/Quality Control ProceduresSample Containers, Handling, and PreservationAncillary Methodology 12TemperatureFoodAcceptability
25、of Test 13Report 14Keywords 15References1.7 The values stated in the International System of Units(SI) (the Modernized Metric System) are to be regarded asstandard.1.8 This standard may involve hazardous materials, opera-tions, and equipment particularly during field operations inturbulent waters or
26、 extreme weather conditions. This standarddoes not purport to address all of the safety concerns, if any,associated with its use. It is the responsibility of the user of thisstandard to establish appropriate safety and health practicesand determine the applicability of regulatory requirementsprior t
27、o use. Specific hazard statements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:3D 1129 Terminology Relating to WaterD 3976 Practice for Preparation of Sediment Samples forChemical AnalysisD 4447 Guide for Disposal of Laboratory Chemicals andSamplesE 724 Guide for Conducting Stati
28、c Acute Toxicity TestsStarting with Embryos of Four Species of SaltwaterBivalve MolluscsE 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 1022 Guide for Conducting
29、 Bioconcentration Tests withFishes and Saltwater Bivalve MollusksE 1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE 1191 Guide for Conducting Life-Cycle Toxicity Testswith Saltwater MysidsE 1192 Guide for Conducting Acute Toxicity Tests onAqueous Ambient Samples
30、and Effluents with Fishes,Macroinvertebrates, and AmphibiansE 1342 Practice for Preservation by Freezing, Freeze-Drying, and Low Temperature Maintenance of Bacteria,Fungi, Protista, Viruses, Genetic Elements, andAnimal andPlant TissuesE 1367 Test Method for Measuring the Toxicity ofSediment-Associat
31、ed Contaminants with Estuarine andMarine InvertebratesE 1391 Guide for Collection, Storage, Characterization, andManipulation of Sediments for Toxicological Testing andfor Selection of Samplers Used to Collect Benthic Inver-tebratesE 1525 Guide for Designing Biological Tests with Sedi-mentsE 1688 Gu
32、ide for Determination of the Bioaccumulation ofSediment-Associated Contaminants by Benthic Inverte-bratesE 1706 Test Method for Measuring the Toxicity ofSediment-Associated Contaminants with Freshwater Inver-tebratesE 1847 Practice for Statistical Analysis of Toxicity TestsConducted Under ASTM Guide
33、linesIEEE/SI 10 American National Standard for Use of theInternational System of Units (SI): The Modern MetricSystem3. Terminology3.1 Definitions:3For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards
34、 volume information, refer to the standards Document Summary page onthe ASTM website.E 2122 02 (2007)23.1.1 The words “must,” “should,” “may,” “can,” and“might,” have very specific meanings in this guide. “Must” isused to express an absolute requirement, that is, to state that atest ought to be desi
35、gned to satisfy the specified condition,unless the purpose of the test requires a different design.“Must” is only used in connection with factors that directlyrelate to the acceptability of the test. “Should” is used to statethat a specified condition is recommended and ought to be metif possible. A
36、lthough violation of one “should” is rarely aserious matter, violation of several will often render the resultsquestionable. Terms such as “is desirable” are used in connec-tion with less important factors. “May” is used to mean “is(are) allowed to,” “can” is used to mean “is (are) able to,” and“mig
37、ht” 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 used in this guide, referto Terminology D 1129, Guide E 729, Terminology E 943, andGuide E 1023.
38、 For an explanation of units and symbols, referto IEEE/SI 10.3.2 Definitions of Terms Specific to This Standard:3.2.1 bioaccumulation factor (BAF)the ratio of tissuechemical residue to chemical concentration in the externalenvironment. BAF is measured at steady state in situationswhere organisms are
39、 exposed from multiple sources (that is,water, sediment, food), unless noted otherwise.3.2.2 bioassayan experiment that includes both an esti-mate of toxicity and an estimate of relative potency.3.2.3 bioavailabilitythe fraction of the total chemicalconcentration in water, on sediment particles, and
40、 on food thatis available for bioaccumulation.3.2.4 biomonitoringuse of living organisms as “sensors”in water or sediment quality surveillance to detect changes inan effluent or water body or to indicate whether aquatic lifemay be endangered.3.2.5 chemical concentrationthe ratio of the weight orvolu
41、me of chemicals to the weight or volume of a test sample.3.2.6 chemical contentmass of chemical per whole ani-mal (for example, g/animal) can be used to normalize theexpression of chemical uptake per unit time by eliminating theeffects of growth on changing tissues masses.3.2.7 chemical fingerprinti
42、ngthe use of specific patternsin the ratios of chemicals accumulated in bivalve tissues toidentify chemical sources; for example, the ratio of PAHalkylated homologs to parent compounds.3.2.8 compartmentalized cagea rigid or flexible meshcage with individual compartments for holding bivalves in acont
43、rolled position so that multiple measurements can be madeon the same individual organism. The compartmentalized cagehelps maximize water flow around individual test organismsand provides even exposure to the test environment.3.2.9 growth dilutiona process whereby the rate of accu-mulation is exceede
44、d by the rate of tissue growth so that whenthe concentration is expressed on mass of chemical per mass oftissue over time, it appears as though depuration or eliminationis occurring because the concentration (g/g) is decreasing.3.2.10 reference stationa station similar to the test sta-tion(s) in phy
45、sical and chemical characteristics and withrelatively little to no contamination by the particular chemi-cal(s) under study. A reference station should ideally containonly background concentrations of chemicals characteristic ofthe region.3.2.11 scope for growthan integrated physiological mea-sure o
46、f the energy status of an organism at a particular time,based on the concept that energy in excess of that required fornormal maintenance will be available for the growth andreproduction of the organism.3.2.12 shell lengththe distance from the tip of the umbo tothe distal valve edge.3.2.13 sitea geo
47、graphical area within a somewhat definedboundary that is being studied. The size of a site is dependenton the extent of suspected perturbation, generally on the orderof 0.1 to 50km2. Part of the vagueness in size is due tovariability in spatial scale and inadequate results from prelimi-nary reconnai
48、ssance survey that clearly define the boundary ofsuspected stressors.3.2.14 steady statethe state in which fluxes of materialmoving bidirectionally across a membrane or boundary be-tween compartments or phases have reached a balance. Anequilibrium between the phases is not necessarily achieved.3.2.1
49、5 stationa specific sampling location or area within asite. The size of a station can vary from a single point with onecage to an area of approximately 10 by 10 m including severalcages. Vagueness in size is due to variability in spatial scaleand experimental design. Several stations in a small geographicarea could comprise a site.3.2.16 tissue loss magnificationthe process whereby thetissue mass is lost during the exposure period and the chemicalmass remains constant over time, so that when the concentra-tion is expressed on mass of chemical per mass of tissue
