1、Designation: E 1711 95 (Reapproved 2003)e1Standard Guide forMeasurement of Behavior During Fish Toxicity Tests1This standard is issued under the fixed designation E 1711; 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 (e) indicates an editorial change since the last revision or reapproval.e1NOTEWarning notes were editorially moved into the text in August 2003.1. Scope1.1 This guide covers some general information on
3、 methodsfor qualitative and quantitative assessment of the behavioralresponses of fish during standard laboratory toxicity tests tomeasure the sublethal effects of exposure to chemical sub-stances. This guide is meant to be an adjunct to toxicity testsand should not interfere with those test procedu
4、res.1.2 Behavioral toxicosis occurs when chemical or otherstressful conditions, such as changes in water quality ortemperature, induce a behavioral change that exceeds thenormal range of variability (1). Behavior includes all of theobservable, recordable, or measurable activities of a livingorganism
5、 and reflects genetic, neurobiological, physiological,and environmental determinants (2).1.3 Behavioral methods can be used in biomonitoring, in thedetermination of no-observed-effect and lowest-observed-effect concentrations, and in the prediction of hazardouschemical impacts on natural populations
6、 (3).1.4 The behavioral methods described in this guide includelocomotory activity, feeding, and social responses, which arecritical to the survival of fish (4).1.5 This guide is arranged as follows:Section NumberScope 1Referenced Documents 2Terminology 3Summary of Guide 4Significance and Use 5Inter
7、ferences 6Safety Precautions 7Responses Measured 8Test Organisms 9Facility 10Qualitative Behavioral Assessment Method 11Quantitative Behavioral Measurements 12Experimental Design 13Calculation of Test Results 14Report 151.6 The values stated in either inch-pound or SI units are tobe regarded as the
8、standard. The values given in parenthesesare for information only.1.7 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 app
9、lica-bility of regulatory limitations prior to use. While some safetyconsiderations are included in this guide, it is beyond the scopeof this guide to encompass all safety requirements necessary toconduct behavioral toxicity tests. Specific hazards statementsare given in Section 7.2. Referenced Docu
10、ments2.1 ASTM Standards:2E 140 Standard Hardness Conversion Tables for MetalsRealtionship Among Brinell Hardness, Vickers Hardness,Rockwell Hardness, Superficial Hardness, Knoop Hard-ness, and Scleroscope HardnessE 729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinve
11、rtebrates, and Amphib-iansE 1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE 1241 Guide for Conducting Early Life-Stage ToxicityTests with FishesE 1383 Guide for Conducting Sediment Toxicity Tests withFreshwater Invertebrates3E 1604 Guide for Behavioral Testing i
12、n Aquatic Toxicology3. Terminology3.1 DefinitionsThe words “must,” “should,” “may,”“can,” and “might” have very specific meanings. “Must” isused to express an absolute requirement, that is, to state that the1This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environm
13、ental Fate and is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved Aug. 10, 2003. Published September 2003. Originallyapproved in 1995. Last previous edition approved in 1995 as E 1711 95.2For referenced ASTM standards, visit the ASTM webs
14、ite, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Withdrawn.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,
15、United States.test ought to be designed to satisfy the specified condition,unless the purpose of the test requires a different design.“Must” is used only in connection with the factors that relatedirectly to the acceptability of the test. “Should” is used to statethat the specified condition is reco
16、mmended and ought to bemet if possible.Although the violation of one “should” is rarelya serious matter, violation of several will often render theresults questionable. Terms such as “is desirable,” “is oftendesirable,” and “might be desirable” are used in connectionwith less important factors. “May
17、” 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.”3.2 Definitions of Terms Specific to This S
18、tandard:3.2.1 aggressive behaviorbehavioral reactions made inresponse to a conspecific resulting in the repulsion of individu-als. Aggressive social behaviors include reactions of ap-proaches; displays of coloration, posture, or body movements;bodily contact; or biting that results in the retreat of
19、 theresponding conspecific or in the initiation of similar responses.3.2.2 feeding behaviora response resulting in the con-sumption of material, including orientation and movementtoward the material, sucking or striking motions directed at thematerial, capture by mouth, spiting or holding, and swall
20、owingof the material.3.2.3 locomotory behaviorneuromuscular responses thatresult in movement of the fishs body or a portion of the bodyin space to cause a change in position or orientation in space,as well as reflexive movements of body parts.3.2.4 schooling or shoaling behaviorresponses of socialat
21、traction that reflect a tendency to remain in the vicinity of aconspecific.4. Summary of Guide4.1 This guide is intended to describe behavioral methodsthat can be applied during routine bioassays. Qualitativebehavioral assessment procedures are intended to providelimited behavioral characterizations
22、 that require minimal facil-ity modifications, equipment, or training and are inexpensive toconduct. Quantitative behavioral assessments are more rigor-ous measurements of behavior and are intended for laboratoriesthat have an interest in behavioral testing and can providelimited modifications of fa
23、cilities and conventional videorecording equipment and limited staff training.4.1.1 Qualitative behavioral screening of spontaneous be-havioral activity provides a broad view of toxicant effectsduring exposure to contaminants. Abnormal behavioral re-sponses observed among fish are documented on a da
24、ily basisusing a behavioral checklist that includes categories of re-sponses such as lack of feeding, lethargic or frenzied activity,abnormal swimming movements or postures, and lack ofresponse or hyperreactivity to external stimuli (5). The behav-ioral aberrations are based on the absence of respon
25、se and onobvious differences from the response of untreated fish. Al-though no attempt is made to quantify the magnitude ofresponse, the consistent observation of response over timeprovides a quantitative measurement of the response. Earlydetection of behavioral abnormalities may warrant quantitativ
26、emeasures of specific behavioral patterns.4.2 Quantitative measurements of locomotory, feeding, andsocial behaviors of fish can be conducted during standardlaboratory exposures, including static, flow-through, sediment,and food exposures from direct observation or overhead videorecordings to determi
27、ne the effects of sublethal exposure (6).These behavioral responses are highly sensitive to sublethalexposure and are relevant to survival (7). Data are obtained todetermine the effects of toxic substances on behavior fromshort (for example, 96 h) or long-term (partial to full life cycle)exposures.5
28、. Significance and Use5.1 Protection of a species requires the prevention of detri-mental effects of chemicals on the survival, growth, reproduc-tion, health, and uses of individuals of that species. Behavioraltoxicity tests provide information concerning the sublethaleffects of chemicals and signal
29、 the presence of toxic testsubstances.5.1.1 The locomotory, feeding, and social responses of fishare adaptive and essential to survival. Major changes in theseresponses may result in a diminished ability to survive, grow,avoid predation, or reproduce and cause significant changes inthe natural popul
30、ation (8). Fish behavioral responses areknown to be highly sensitive to environmental variables as wellas toxic substances.5.2 Results from behavioral toxicity tests may be useful formeasuring injury resulting from the release of hazardousmaterials (9).5.3 Behavioral responses can also be qualitativ
31、ely assessedin a systematic manner during toxicity tests to discern trends insublethal contaminant effects (5).5.4 The assessment of locomotory, feeding, and socialbehaviors is useful for monitoring effluents and sediments fromcontaminated field sites as well as for defining no-effectconcentrations
32、in the laboratory or under controlled fieldconditions. Such behavioral modifications provide an index ofsublethal toxicity and also indicate the potential for subsequentmortality.5.5 Behavioral toxicity data can be used to predict theeffects of exposure likely to occur in the natural environment(10)
33、.5.6 Results from behavioral toxicity tests might be animportant consideration when assessing the hazard of materialsto aquatic organisms. Such results might also be used whenderiving water quality criteria for fish and aquatic invertebrateorganisms.5.7 Results from behavioral toxicity tests can be
34、used tocompare the sensitivities of different species, the relativetoxicity of different chemical substances on the same organism,or the effect of various environmental variables on the toxicityof a chemical substance.5.8 Results of behavioral toxicity tests can be useful inguiding decisions regardi
35、ng the extent of remedial actionneeded for contaminated aquatic and terrestrial sites.5.9 The behavioral characteristics of a particular organismneed to be understood and defined before a response can beused as a measure of toxicity (11). Swimming, feeding, andE 1711 95 (2003)e12social behavior vari
36、es among species as well as among lifestages within a species; the most effective test methods aretherefore those tailored to a particular life stage of a singlespecies. The range of variability of any behavioral response ofunexposed organisms is influenced by genetic, experiential,physiological, an
37、d environmental factors. It is thus important toavoid selecting test organisms from populations that may varyin these factors.5.10 Results of behavioral toxicity tests will depend on thebehavioral response measured, testing conditions, water qual-ity, species, genetic strain, life stage, health, and
38、 condition oftest organisms. The behavioral response may therefore beaffected by the test environment.5.11 No numerical value or range of values has been definedas the norm for swimming, feeding, or social behavior for anyfish; the detection of abnormal activity is therefore based oncomparisons of t
39、he responses of exposed fish, either withactivity measured during a baseline or pre-exposure period orobservations of fish under a control treatment (10).5.12 These measures are incorporated readily into standardtoxicity test protocols, with minimal stress to the test organism.6. Interferences6.1 A
40、number of factors can suppress, elicit, or alterlocomotory, feeding, and social responses and thus influencebehavioral test results and complicate data interpretation. Thefollowing factors should be considered when measuring be-havioral responses during toxicity tests:6.1.1 The pretest handling of t
41、est organisms resulting fromcollection, transfer, and maintenance of the culture environ-ment can affect the response observed during exposure to toxicsubstances.6.1.2 The health, nutritional state, and physical condition ofthe organism can influence the test.6.1.3 Behavioral responsiveness may vary
42、 by species, ge-netic strain, population, gender, and developmental stage of theorganism.6.1.4 Prior exposure to hazardous materials, environmentalstresses, and pathogens can affect behavioral responses.6.1.5 Social status, such as the dominance or sex of theindividuals tested, and experiential fact
43、ors, such as priorexperience with the predator or prey species, can influence thebehavioral response. Individuals tested in isolation may re-spond differently from when tested in groups.6.1.6 Cyclical changes (circadian, seasonal, annual, hor-monal, and reproductive) in behavioral responses can occu
44、r.6.1.7 The behavioral response can be affected by the appa-ratus design and procedural sequence of the measurementmethod.6.1.8 Behavioral responses will vary according to the extentto which test organisms acclimate to the physical variables ofthe testing environment, including water quality, temper
45、ature,water flow, light, cover, and substrate, as well as their recoveryfrom handling, acceptance of diet, and adjustment to noveltesting chambers.6.1.9 It is very important to eliminate disturbances to thetest system, such as vibrations, slamming doors, castingshadows, abrupt changes in lighting, o
46、r water flow, that mayfrighten the fish or disrupt ongoing activity.6.1.10 Behavioral responses to toxic substances may sub-side over time.6.1.11 Precise, objective, operational definitions of behav-ioral endpoints measured during toxicity tests are required.6.1.12 Generally, excessive mortality amo
47、ng controls (seeGuides E 729 and E 1241), high variability in the behavioralresponse of controls, disease, or variation in water quality orexperimental parameters beyond acceptable limits, and incon-sistent visualization of the organism are the basis for rejectinga behavioral test. The criteria for
48、such limits will varydepending on the substance, species, and response being tested,as well as the objectives of the study. Guide E 1604 should beconsulted regarding the acceptability of behavioral test results.7. Safety Precautions7.1 Many substances may pose health risks to humans ifadequate preca
49、utions are not taken. Information on toxicity tohumans, recommended handling procedures, and the chemicaland physical properties of the test material should be studiedand all personnel informed before an exposure is initiated.(WarningSpecial procedures might be necessary with radio-labeled test materials and with test materials that are, or aresuspected of being, carcinogenic.)7.2 Many materials can affect humans adversely if precau-tions are inadequate. Contact with test material, sediments, andwater should be minimized. Where appropriate, protectivegloves, laboratory coats,
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