1、Designation: E2122 02 (Reapproved 2013)Standard Guide forConducting In-situ Field Bioassays With Caged Bivalves1This standard is issued under the fixed designation E2122; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of
2、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 describes procedures for conducting con-trolled experiments with caged bivalves under field conditions.The
3、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 US E
4、PAecological 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 associat
5、ed 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 experimentati
6、on 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 and
7、time within a clearlydefined exposure period. Chemical exposure can be estimatedby measuring the concentration of chemicals in water,sediment, or bivalve tissues, and effects can be estimated withsurvival, growth, and other sublethal end points (3). Althougha number of assessments have been conducte
8、d 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 in t
9、issue 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 pote
10、ncy 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 field st
11、udies. 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, dissolvedand sus
12、pended 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 facultativedeposit-fee
13、ding 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 beenconducted in
14、 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 in mar
15、ine, 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 bioav
16、ailability 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 mass) o
17、f chemical per animal (for example,g/animal), rate of uptake, or bioaccumulation factor (BAF, theratio between the concentration of a chemical in bivalve tissuesand the concentration in the external environment, including1This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffect
18、s and Environmental Fate and is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved March 1, 2013. Published March 2013. Originallyapproved in 2001. Last previous edition approved in 2007 as E2122 02(2007).DOI: 10.1520/E2122-02R13.2The boldfa
19、ce numbers in parentheses refer to references at the end of thisstandard.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1water, sediment, and food). Tissue chemistry results can onlybe used to calculate a BAF because caged bivalves in
20、 the fieldare exposed to 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 pro
21、cedures might be justi-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 obta
22、ined using modified and 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 U
23、se 5Interferences 6Hazards 7Experimental Design 8Apparatus 9FacilitiesConstruction MaterialsCagesTest Organisms 10SpeciesCommonly Used TaxaSize and Age of Test OrganismsSourceNumber of SpecimensCollectionHandlingHoldingAnimal QualityField Procedures 11Test Initiation: PresortFinal Measurements and D
24、istributionAttachment of 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 12Temper
25、atureFoodAcceptability of Test 13Report 14Keywords 15References1.7 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.1.8 This standard may involve hazardous materials,operations, and equipment particularly during field opera-tions
26、 in turbulent waters or extreme weather conditions. Thisstandard does not purport to address all of the safety concerns,if any, associated with its use. It is the responsibility of the userof this standard to establish appropriate safety and healthpractices and determine the applicability of regulat
27、ory require-ments prior to use. Specific hazard statements are given inSection 7.2. Referenced Documents2.1 ASTM Standards:3D1129 Terminology Relating to WaterD3976 Practice for Preparation of Sediment Samples forChemical AnalysisD4447 Guide for Disposal of Laboratory Chemicals andSamplesE724 Guide
28、for Conducting Static Acute Toxicity TestsStarting with Embryos of Four Species of SaltwaterBivalve MolluscsE729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE943 Terminology Relating to Biological Effects and Envi-ronmental FateE1022 Gui
29、de for Conducting Bioconcentration Tests withFishes and Saltwater Bivalve MollusksE1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE1191 Guide for Conducting Life-Cycle Toxicity Tests withSaltwater MysidsE1192 Guide for Conducting Acute Toxicity Tests on Aque-ous
30、Ambient Samples and Effluents with Fishes,Macroinvertebrates, and AmphibiansE1342 Practice for Preservation by Freezing, Freeze-Drying,and Low Temperature Maintenance of Bacteria, Fungi,Protista, Viruses, Genetic Elements, and Animal and PlantTissues (Withdrawn 2011)4E1367 Test Method for Measuring
31、the Toxicity of Sediment-Associated Contaminants with Estuarine and Marine In-vertebratesE1391 Guide for Collection, Storage, Characterization, andManipulation of Sediments for Toxicological Testing andfor Selection of Samplers Used to Collect Benthic Inver-tebratesE1525 Guide for Designing Biologic
32、al Tests with SedimentsE1688 Guide for Determination of the Bioaccumulation ofSediment-Associated Contaminants by Benthic Inverte-bratesE1706 Test Method for Measuring the Toxicity of Sediment-Associated Contaminants with Freshwater InvertebratesE1847 Practice for Statistical Analysis of Toxicity Te
33、stsConducted Under ASTM GuidelinesIEEE/SI 10 American National Standard for Use of theInternational System of Units (SI): The Modern MetricSystem3For 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.4The last approved version of this historical standard is referenced onwww.astm.org.E2122 02 (2013)23. Terminology3.1 Definitions:3.1.1 The words “must,” “should,” “may,” “can,” and“might,” have very specific meaning
35、s in this guide. “Must” isused to express an absolute requirement, that is, to state that atest ought to be designed 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 o
36、f the test. “Should” is used to statethat a specified condition is recommended and ought to be metif possible. Although 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
37、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 as a synonym for either “may” or “can.”3.1.2 For definitions
38、 of other terms used in this guide, referto Terminology D1129, Guide E729, Terminology E943, andGuide E1023. For an explanation of units and symbols, refer toIEEE/SI 10.3.2 Definitions of Terms Specific to This Standard:3.2.1 bioaccumulation factor (BAF)the ratio of tissuechemical residue to chemica
39、l concentration in the externalenvironment. BAF is measured at steady state in situationswhere organisms are 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 poten
40、cy.3.2.3 bioavailabilitythe fraction of the total chemicalconcentration in water, on sediment particles, and 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
41、to indicate whether aquatic lifemay be endangered.3.2.5 chemical concentrationthe ratio of the weight orvolume of chemicals to the weight or volume of a test sample.3.2.6 chemical contentmass of chemical per whole animal(for example, g/animal) can be used to normalize the expres-sion of chemical upt
42、ake per unit time by eliminating the effectsof growth on changing tissues masses.3.2.7 chemical fingerprintingthe 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 c
43、ompartmentalized cagea rigid or flexible meshcage with individual compartments for holding bivalves in acontrolled position so that multiple measurements can be madeon the same individual organism. The compartmentalized cagehelps maximize water flow around individual test organismsand provides even
44、exposure to the test environment.3.2.9 growth dilutiona process whereby the rate of accu-mulation is exceeded 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 th
45、e concentration (g/g) is decreasing.3.2.10 reference stationa station similar to the test sta-tion(s) in physical and chemical characteristics and withrelatively little to no contamination by the particular chemi-cal(s) under study. A reference station should ideally containonly background concentra
46、tions of chemicals characteristic ofthe region.3.2.11 scope for growthan integrated physiological mea-sure of 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 th
47、e organism.3.2.12 shell lengththe distance from the tip of the umbo tothe distal valve edge.3.2.13 sitea geographical 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 t
48、he vagueness in size is due tovariability in spatial scale and inadequate results from prelimi-nary reconnaissance 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 compa
49、rtments or phases have reached a balance. Anequilibrium between the phases is not necessarily achieved.3.2.15 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 re
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