1、Designation: E 1463 92 (Reapproved 2004)Standard Guide forConducting Static and Flow-Through Acute Toxicity TestsWith Mysids From the West Coast of the United States1This standard is issued under the fixed designation E 1463; the number immediately following the designation indicates the year oforig
2、inal 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 guide describes procedures for obtaining dataconcerning t
3、he adverse effects of a test material (not food)added to marine and estuarine waters on certain species ofmarine and estuarine mysids during 96 h of continuousexposure. Juvenile mysids used in these tests are taken fromcultures shortly after release from the brood and exposed tovarying concentration
4、s of a toxicant in static or flow-throughconditions. These procedures will be useful for conductingtoxicity tests with other species of mysids, although modifica-tions might be necessary.1.2 Modifications of these procedures might be justified byspecial needs or circumstances. Although using appropr
5、iateprocedures is more important than following prescribed pro-cedures, results of tests conducted using unusual proceduresare not likely to be comparable to results of many other tests.Comparisons of results obtained using modified and unmodi-fied versions of these procedures might provide useful i
6、nfor-mation concerning new concepts and procedures for conduct-ing acute tests with other species of mysids.1.3 The procedures given in this guide are applicable tomost chemicals, either individually or in formulations, com-mercial products, and known or unknown mixtures. Withappropriate modificatio
7、ns these procedures can be used toconduct acute tests on factors such as temperature, salinity, anddissolved oxygen. These procedures can also be used to assessthe toxicity of potentially toxic discharges such as municipalwastes, oil drilling fluids, produced water from oil wellproduction, and other
8、 types of industrial wastes.1.4 Results of acute toxicity tests with toxicants experimen-tally added to salt and estuarine waters should usually bereported in terms of a LC50 (median lethal concentration).1.5 This guide is arranged as follows:SectionReferenced Documents 2Terminology 3Summary of Guid
9、e 4Significance and Use 5Apparatus 6Facilities 6.1Construction Materials 6.2Metering Systems 6.3Test Chambers 6.4Cleaning 6.5Acceptability 6.6Safety Precautions 7Dilution Water 8Requirements 8.1Source 8.2Treatment 8.3Characterization 8.4Test Material 9General 9.1Stock Solution 9.2Test Concentrations
10、 9.3Test Organisms 10Species 10.1Age 10.2Source 10.3Brood Stock 10.4Food 10.5Handling 10.6Harvesting Young 10.7Quality 10.8Procedure 11Experimental Design 11.1Dissolved Oxygen 11.2Temperature 11.3Loading 11.4Salinity 11.5Light 11.6Beginning of Test 11.7Feeding 11.8Duration of Test 11.9Biological Dat
11、a 11.10Other Measurements 11.11Analytical Methodology 12Acceptability of Test 13Interpretation of Results 14Report 15AppendixesHolmesimysis costata X1Neomysis mercedis X21.6 The values stated in SI units are to be regarded as thestandard.1This guide is under the jurisdiction of ASTM Committee E47 on
12、 BiologicalEffects and Environmental Fate and is the direct responsibility of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved April 1, 2004. Published April 2004. Originallyapproved in 1992. Last previous edition approved in 1998 as E 1463 92 (1998).1Copyright ASTM I
13、nternational, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.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
14、 practices and determine the applica-bility of regulatory limitations prior to use. Specific precau-tionary statements are given in Section 7.2. Referenced Documents2.1 ASTM Standards:2E 380 Practice for Use of the International System of Units(SI) (the Modernized Metric System)3E 729 Guide for Cond
15、ucting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates, and Amphib-iansE 943 Terminology Relating to Biological Effects and En-vironmental FateE 1023 Guide for Assessing the Hazard of a Material toAquatic Organisms and Their UsesE 1191 Guide for Conducting Life-Cycle Toxicity T
16、estswith Saltwater MysidsE 1192 Guide for Conducting Acute Toxicity Tests onAqueous Ambient Samples and Effluents with Fishes,Macroinvertebrates, and AmphibiansE 1203 Practice for Using Brine Shrimp Nauplii as Food forTest Animals in Aquatic Toxicology3. Terminology3.1 The terms “must,” “should,” “
17、may,” “can,” and “might”have very specific meanings in this guide. “Must” is used toexpress the strongest possible recommendation, just short of anabsolute requirement, that is, to state that this test ought to bedesigned to satisfy the specific condition, unless the purpose ofthe test requires a di
18、fferent design. “Must” is only used inconnection with factors that directly relate to the acceptabilityof the test (see Section 13).3.1.1 “Should” is used to state that the specific condition isrecommended and ought to be met if possible. Althoughviolation of one “should” is rarely a serious matter,
19、 violation ofseveral will often render the results questionable. Terms suchas “is desirable.” “is often desirable,” and “might be desirable”are used in connection with less important factors.3.1.2 “May” is used to mean “is (are) allowed to,” “can” isused to mean “is (are) able to,” and “might” is us
20、ed to mean“could possibly.” Thus the classic distinction between “may”and “can” is preserved, and “might” is never used as asynonym for either “may” or “can.”3.2 Definitions:3.2.1 LC50the statistically or graphically derived bestestimate of the concentration of a toxicant added to an aqueoussolution
21、 that results in the death of 50 % of the test organismswithin the test period (see Definition E 943).3.2.2 For definitions of other terms used in this guide, referto Guide E 729, Terminology E 943, Guides E 1191 andE 1192, and Practice E 1203. For an explanation of units andsymbols, refer to Practi
22、ce E 380.4. Summary of Guide4.1 The toxicity of a substance in marine or estuarine waterscan be determined through a 96-h acute static test or aflow-through test. Static tests may be conducted in glass 2-Lfinger bowls or 350-mL finger bowls. Either size finger bowlprovides a large surface-to-volume
23、ratio and ample horizontalspace to minimize cannibalism. The dishes should be coveredand aerated. Whichever static technique is used, specific dataon the concentration of test material are obtained and analyzedto determine the effect(s) of the toxicant on survival. In theflow-through acute technique
24、, the test solution flows throughthe test chamber on a once-through basis throughout the test.The flow-through test is considered to be more representativeof actual field conditions, but not all laboratories have thecapabilities of conducting this type of test.5. Significance and Use5.1 Mysids are a
25、n important component of both the pelagicand epibenthic community. They are preyed upon by manyspecies of fish, birds, and larger invertebrate species, and theyare predators of smaller crustaceans and larval stages ofinvertebrates. In some cases, they feed upon algae. Mysids aresensitive to both org
26、anic and inorganic toxicants (1).4Theecological importance of mysids, their wide geographicaldistribution, ability to be cultured in the laboratory, andsensitivity to contaminants make them appropriate acute tox-icity test organisms.5.2 An acute toxicity test is conducted to obtain informationconcer
27、ning the immediate effects of a short-term exposure to atest material on a test organism under specified experimentalconditions. An acute toxicity test provides data on the short-term effects that are useful for comparisons to other species butdoes not provide information on delayed effects.5.3 Resu
28、lts of acute toxicity tests can be used to predictacute effects likely to occur on aquatic organisms in fieldconditions except that mysids might avoid exposure whenpossible.5.4 Results of acute toxicity tests might be used to comparethe acute sensitivities of different species and the acutetoxicitie
29、s of different test materials, and to study the effects ofvarious environmental factors on results of such tests.5.5 Results of acute toxicity tests might be an importantconsideration when assessing the hazards of materials toaquatic organisms (see Guide E 1023) or when deriving waterquality criteri
30、a for aquatic organisms (2).5.6 Results of acute toxicity tests might be useful forstudying biological availability of, and structure activity rela-tionships between test materials.5.7 Results of acute toxicity tests will depend, in part, on thetemperature, quality of the food, condition of test org
31、anisms,test procedures, and other factors.2For referenced ASTM standards, visit the ASTM website, 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.4The bol
32、dface numbers given in parentheses refer to a list of references at theend of this guide.E 1463 92 (2004)26. Apparatus6.1 FacilitiesAquaria or tanks containing either clean(uncontaminated) natural sea water or reconstituted sea water(see 8.2) should be used for holding mysids after fieldcollection a
33、nd prior to a test. Both static-recirculating andflow-through holding systems have been used successfully (1).Cultures of Holmesimysis costata have not been reported formedia of reconstituted sea water. The holding tanks and anyarea used for manipulating live mysids should be located in aroom or spa
34、ce separated from that in which toxicity tests are tobe conducted. The sea water should be monitored periodicallyto ensure a constant salinity. The holding tanks, water supply,or the room in which they are kept should be equipped withtemperature control. Aeration should be provided to ensure thatthe
35、 concentration of dissolved oxygen is greater than 60 % ofsaturation as well as adequate water circulation in the tanks. Atiming device should be used to provide a 16-h light and an 8-hdark photoperiod. A 15 to 30-min transition period (3) whenlights go on might be desirable to reduce the possibilit
36、y oforganisms being stressed by large, sudden increases in lightintensity. A transition period when the lights go off might alsobe desirable.6.2 Construction MaterialsEquipment and facilities thatcontact stock solutions, test solutions, or any water into whichtest organisms will be placed should not
37、 contain substancesthat can be leached or dissolved by aqueous solutions inamounts that adversely affect test organisms. In addition,equipment and facilities that contact stock or test solutionsshould be chosen to minimize sorption of test materials fromwater. Glass, Type 316 stainless steel, nylon,
38、 and fluorocarbonplastics should be used whenever possible to minimize disso-lution, leaching, and sorption, except that stainless steel shouldnot be used in tests on metals in salt water. Concrete and rigidplastics may be used for holding tanks and in the water-supplysystem, but they should be soak
39、ed, preferably in flowingdilution water, for a week or more before use (4). Cast iron pipeshould not be used with salt water and probably should not beused in a fresh water-supply filter system because colloidal ironwill be added to the dilution water and strainers will be neededto remove rust parti
40、cles. A specially designed system is usuallynecessary to obtain salt water from a natural water source (5).Brass, copper, lead, galvanized metal, and natural rubbershould not contact dilution water, stock solutions, or testsolutions before or during the test. Items made of neoprenerubber or other ma
41、terials not mentioned previously should notbe used unless it has been shown that their use will notadversely affect either survival, growth, or reproduction ofmysids.6.3 Metering System:6.3.1 For flow-through tests, the metering system should bedesigned to accommodate the type and concentration(s) o
42、f thetest material and the necessary flow rates of test solutions. Thesystem should permit the mixing of test material with dilutionwater immediately before entrance to the test chambers andpermit the supply of the selected concentration(s) of testmaterial (see 9.1.3 and 11.7.5) in a reproducible fa
43、shion.Various metering systems, using different combinations ofsyringes, “dipping birds,” siphons, pumps, saturators, sole-noids, valves, and so forth, have been used successfully tocontrol the concentrations of test material in, and the flow ratesof, test solutions (6). Proportional diluters (7) us
44、e an intermit-tent flow design and various devices for metering the testmaterial (8). Continuous-flow metering systems are also avail-able, as are systems that prepare the different test solutionsindependently of each other (6).6.3.2 The metering system should be calibrated before andafter the test
45、by determining the flow rate through each testchamber and by measuring either the concentration of testmaterial in each test chamber or the volume of solution used ineach portion of the metering system. The general operation ofthe metering system should be visually checked daily in themorning and af
46、ternoon throughout the test. The meteringsystem should be adjusted during the test if necessary.6.3.3 The flow rate through each test chamber should be atleast five-volume additions per 24 h. It is usually desirable toconstruct the metering system so that it can provide at leastten-volume additions
47、per 24 h in case the loading is high (see11.4) or there is rapid loss of test material due to microbialdegradation, hydrolysis, oxidation, photolysis, reductions,sorption, or volatilization. At any particular time during thetest, the flow rates through any two test chambers should notdiffer by more
48、than 10 %.6.4 Test Chambers:6.4.1 In a toxicity test with aquatic organisms, test chambersare defined as the smallest physical units between which thereare no water connections. Screens and cups may be used tocreate two or more compartments within each chamber butsuch compartments are not replicate
49、experimental units. There-fore, the test solution can be in contact with the test organismsin the compartments within a test chamber, but, by definition,cannot flow from one chamber to another. Because the solutioncan flow from one compartment to another in the same testchamber, the temperature, concentration of test material, ortypes of pathogens and extraneous contaminants are likely tobe more similar between compartments in the same testchamber than between compartments in different test chambersin the same treatment. Chambers should be covered to keep outextraneous contaminant
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