1、Designation: E1463 92 (Reapproved 2012)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 E1463; the number immediately following the designation indicates the year oforigin
2、al adoption or, in the case of revision, the year of 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 obtaining dataconcerning the
3、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 concentrations o
4、f 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 appropriat
5、eprocedures is more important than following prescribedprocedures, 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 infor-
6、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 modifications th
7、ese 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 type
8、s 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 Guide 4Si
9、gnificance 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 9.3T
10、est 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 Data 11.
11、10Other Measurements 11.11Analytical Methodology 12Acceptability of Test 13Interpretation of Results 14Report 15AppendixesHolmesimysis costata X1Neomysis mercedis X21This guide is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fate and is the direct responsibilit
12、y of SubcommitteeE47.01 on Aquatic Assessment and Toxicology.Current edition approved Dec. 1, 2012. Published January 2013. Originallyapproved in 1992. Last previous edition approved in 2004 as E1463 92 (2004).DOI: 10.1520/E1463-92R12.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700,
13、 West Conshohocken, PA 19428-2959. United States11.6 The values stated in SI units are to be regarded as thestandard.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-priat
14、e safety and health 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:2E729 Guide for Conducting Acute Toxicity Tests on TestMaterials with Fishes, Macroinvertebrates,
15、 and Amphib-iansE943 Terminology Relating to Biological Effects and Envi-ronmental FateE1023 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
16、-ous Ambient Samples and Effluents with Fishes,Macroinvertebrates, and AmphibiansE1203 Practice for Using Brine Shrimp Nauplii as Food forTest Animals in Aquatic ToxicologyIEEE/ASTM SI 10 American National Standard for Use ofthe International System of Units (SI): The Modern MetricSystem3. Terminolo
17、gy3.1 The terms “must,” “should,” “ 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
18、the purpose ofthe test requires a different 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
19、“should” is rarely a serious matter, 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 “
20、is (are) able to,” and “might” is used 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
21、toxicant added to an aqueoussolution that results in the death of 50 % of the test organismswithin the test period (see Definition E943).3.2.2 For definitions of other terms used in this guide, referto Guide E729, Terminology E943, Guides E1191 and E1192,and Practice E1203. For an explanation of uni
22、ts and symbols,refer to IEEE/ASTM SI 10.4. 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 bowlprov
23、ides a large surface-to-volume 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
24、theflow-through acute technique, 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. Signi
25、ficance and Use5.1 Mysids are an 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.
26、Mysids aresensitive to both organic and inorganic toxicants (1).3Theecological 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 conduc
27、ted to obtain informationconcerning 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 informat
28、ion on delayed effects.5.3 Results 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
29、 species and the acutetoxicities 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 E1023) or when
30、 deriving waterquality criteria for aquatic organisms (2).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.3Th
31、e boldface numbers given in parentheses refer to a list of references at theend of this guide.E1463 92 (2012)25.6 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 w
32、ill depend, in part, on thetemperature, quality of the food, condition of test organisms,test procedures, and other factors.6. 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
33、 fieldcollection and 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 locat
34、ed in aroom or space 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
35、to ensure thatthe 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. A15 to 30-min transition period (3) whenlights go on might be desirable to reduc
36、e the possibility 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 p
37、laced should not 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 stainle
38、ss steel, nylon, and fluorocarbonplastics should be used whenever possible to minimizedissolution, leaching, and sorption, except that stainless steelshould not be used in tests on metals in salt water. Concrete andrigid plastics may be used for holding tanks and in thewater-supply system, but they
39、should be soaked, preferably inflowing dilution water, for a week or more before use (4). Castiron pipe should not be used with salt water and probablyshould not be used in a fresh water-supply filter system becausecolloidal iron will be added to the dilution water and strainerswill be needed to rem
40、ove rust particles. A specially designedsystem is usually necessary to obtain salt water from a naturalwater source (5). Brass, copper, lead, galvanized metal, andnatural rubber should not contact dilution water, stocksolutions, or test solutions before or during the test. Items madeof neoprene rubb
41、er or other materials not mentioned previouslyshould not be used unless it has been shown that their use willnot adversely 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 conc
42、entration(s) of 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 r
43、eproducible fashion.Various metering systems, using different combinations ofsyringes, “dipping birds,” siphons, pumps, saturators,solenoids, valves, and so forth, have been used successfully tocontrol the concentrations of test material in, and the flow ratesof, test solutions (6). Proportional dil
44、uters (7) use an intermit-tent flow design and various devices for metering the testmaterial (8). Continuous-flow metering systems are alsoavailable, as are systems that prepare the different test solutionsindependently of each other (6).6.3.2 The metering system should be calibrated before andafter
45、 the test 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 themorn
46、ing and afternoon 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
47、additions 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 notdiffe
48、r by more 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
49、replicate experimental units.Therefore, the test solution can be in contact with the testorganisms in the compartments within a test chamber, but, bydefinition, cannot flow from one chamber to another. Becausethe solution can flow from one compartment to another in thesame test chamber, the temperature, concentration of testmaterial, or types of pathogens and extraneous contaminantsare likely to be more similar between compartments in thesame test chamber than between compartments in different testchambers in the same treatment. Chambers should be coveredto keep out extraneous co
