1、Designation: E 857 05e1Standard Practice forConducting Subacute Dietary Toxicity Tests with AvianSpecies1This standard is issued under the fixed designation E 857; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last re
2、vision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.e1NOTESection 1.4 was editorially revised in February 2006.1. Scope1.1 This practice describes a procedure for determining thesubacute d
3、ietary toxicity of a test substance administered tobirds in their daily diet. The LC50value time to mortality andslope of the dose response curve may also be derived.1.2 This practice is applicable to substances that can bemixed uniformly into the diet.1.3 This practice is intended primarily to be u
4、sed with theyoung of the following species: northern bobwhite (Colinusvirginianus), Japanese quail (Coturnix japonica), mallard(Anas platyrhynchos), and ring-necked pheasant (Phasianuscolchicus). Other species or age groups, for example, withwild-trapped birds, may be used with appropriate husbandry
5、modifications to the practice.1.4 This standard is used routinely to address avian regula-tory testing requirements. Modifications to the proceduresdescribed in this standard have been proposed and are beingevaluated to better address the needs of the latest risk assess-ment procedures. Specifically
6、, the latest procedures call forindividual bird feed consumption measurements so that a moreprecise dose can be determined. While such procedures mayreplace procedures described in the current standard, there isno certainty that the newest procedures will work as antici-pated, and validation is not
7、complete. Therefore, the currentguideline has utility prior to validation and acceptance of amodified standard.1.5 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 safe
8、ty and health practices and determine the applica-bility of regulatory limitations prior to use. For specificprecautionary statements see Section 6.2. Referenced Documents2.1 ASTM Standards:2IEEE/ASTM SI 10 American National Standard for Use ofthe International System of Units (SI): The Modern Metri
9、cSystem3. Terminology3.1 Definitions of Terms Specific to This Standard:3.1.1 LC50the statistically derived estimate of the concen-tration of a test substance in the diet that would be expected tocause 50 % mortality to the test population under the specifiedtest conditions.3.1.2 concentrationthe we
10、ight of test substance per unitweight of diet.3.1.3 substance or test substancethe element, chemicalcompound, formulation, known mixture, or material mixed indiets and fed to birds for the purpose of determining an LC50.3.1.4 negative controla group of birds maintained underconditions identical to t
11、he test birds except for the absence ofthe test substance in their diet.3.1.5 positive controla group of birds maintained underconditions identical to the test birds except for the replacementof the test substance in the diet with a substance known to elicita consistent toxic response.3.2 Abbreviati
12、ons:Units and Symbols:3.2.1 Refer to IEEE/ASTM SI 10.4. Summary of Practice4.1 This practice describes how to determine the subacutedietary toxicity of a test substance when administered to birdsin their daily diet. The median lethal concentration (LC50)inthe diet is a measure of a specific toxic ef
13、fect (that is, lethality).1This practice is under the jurisdiction of ASTM Committee E47 on BiologicalEffects and Environmental Fate and is the direct responsibility of SubcommitteeE47.02 on Terrestrial Assessment and Toxicology.Current edition approved Oct. 1, 2005. Published October 2005. Original
14、lyapproved in 1981. Last previous edition approved in 1997 as E 857 87(1997).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 ont
15、he ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.The LC50has been used as a comparable index of toxicity.However, other expressions of toxicity also may be appropri-ate.4.2 Groups of birds of the same species are fed d
16、ietscontaining a test substance or mixture of substances at selectedconcentrations for 5 days. This is followed by a minimum of 3days (or for as long as the birds continue to exhibit toxic signs)on untreated food. The test substance is mixed into the diets,usually in a geometric series of concentrat
17、ions.4.3 General observations of the signs of toxicity and theacceptance of the test substance in the diet also must bereported.4.4 Concurrent negative controls must be maintainedthroughout the test. A positive control also may be used.5. Significance and Use5.1 This practice provides a means of mea
18、suring the sus-ceptibility of an avian species to a test substance in its dietunder controlled conditions. The LC50obtained in this test is aconditional measure of subacute toxicity because consumptionis voluntary, and because the dietary route may introducemetabolic transformations of the test subs
19、tance that might beabsent in other exposure techniques.5.2 Use of this practice contributes to the evaluation of thehazards of chemicals to birds because exposure is analogous tomost field exposures, that is, through dietary intake.5.3 The use of this practice allows for observation of signsof toxic
20、ity in addition to mortality.5.4 The dose-response curve provides additional informa-tion about the response of birds to a test substance.5.5 This practice can be used to study the effects of testsubstances in combination in order to simulate situations wherebirds may be exposed to more than one sub
21、stance simulta-neously (1).35.6 This practice provides one basis for deciding whetheradditional toxicity testing should be conducted with birds.6. Precautions6.1 Contact with all test substances, solutions, and mixeddiets should be minimized with appropriate protective clothing,gloves, eye protectio
22、n, etc. The use of fume hoods andincreased ventilation in test rooms is necessary when handlingvolatile substances. Mammalian toxicity and special handlingprocedures should be known before this practice is used.6.2 Disposal of excess test substances, solutions, mixeddiets, excreta, and treated birds
23、 should be done with consider-ation for health and environmental safety, and in accordancewith all federal, state, and local regulations.6.3 Cleaning and rinsing of glassware, feeders, and otherequipment with volatile solvents should be performed only inwell-ventilated areas.6.4 Periodic medical exa
24、minations should be considered forall personnel caring for birds or handling test substances.7. Facilities7.1 Species requirements will vary, but pens and cagesshould include adequate room, clean food and water, heatedareas for young birds, and protection from excessive distur-bance. Space requireme
25、nts have not been standardized forspecies normally used in this test. However, adherence to thegeneral guidelines and principles found in the National Insti-tutes of Health and the National Academy of Science publica-tions (2, 3, 4) in addition to literature published on individualspecies should pro
26、vide a basis for a humane approach to spacerequirements. Pens or cages must be placed so as to preventcross-contamination (5).7.2 Construction materials in contact with birds should notbe toxic, nor be capable of adsorbing or absorbing testsubstances. Materials that can be dissolved by water orloose
27、ned by pecking should not be used. Stainless or galva-nized steel, or materials coated with plastics are acceptable, butother construction materials may also be useful. Any materialor pen shape is acceptable provided the birds are able to moveabout freely and that pens can be kept clean.7.3 Ventilat
28、ion, photoperiod, and relative humidity require-ments vary little among test species, and these factors areparticularly critical to the well-being of young birds. Relativehumidity should be maintained at 45 to 70 %. Higher humidi-ties may be appropriate for waterfowl. Photoperiod should bea minimum
29、of 14 h of light. The amount and duration of heatfor brooding is species specific (6, 7). A temperature gradientfrom approximately 38C to approximately 22C from anappropriate heat source should be established in brooders inorder to allow the birds to seek a proper temperature. Ventila-tion should fo
30、llow guidelines in Guide for the Care and Use ofLaboratory Animals. Ventilation should be sufficient to supply10 to 15 air changes per hour (2).8. Diets8.1 Dietary requirements vary according to the species andage of the test birds. Any unmedicated commercial diet thatmeets the minimum nutritional s
31、tandards of the test species (8)is sufficient.8.2 Contaminated feed may compromise a study (9, 10, 11);therefore, feed should be analyzed periodically to identifybackground contaminants. Analysis may be especially impor-tant if the substance being tested is known or suspected ofsynergistic or antago
32、nistic action with possible contaminants.Maximum allowable levels of heavy metals, pesticides, andother contaminants in feed have not been established.8.3 Test diets should always be fresh and clean. Thefrequency that the diet is changed during a study is dependentupon the physical and chemical prop
33、erties of the test substance,and the speed with which a test animal contaminates the feedwith fecal matter or water, or both.8.4 Test diets should be fed ad libitum.8.5 Feed should not be used past its normal shelf life(usually 90 days).8.6 Treated test diets should be stored so as to maintain thest
34、ability of the test substance in the diet.3The boldface numbers in parentheses refer to the list of references at the end ofthis practice.E85705e129. Test Substance and Diet Preparation9.1 Knowledge of the physical, chemical, and biologicalproperties of the test substance is important in test dietpr
35、eparation.9.2 Test diets can be prepared by mixing the test substancedirectly into the feed or by dissolving or suspending the testsubstance in a solvent or carrier prior to mixing with the feed.The use of solvents or carriers may be necessary to achieve auniform mix of the test substance in the fee
36、d.9.3 The test substance is uniformly mixed into the diet. Thephysical and chemical properties of a test substance may causevariation in test diet concentrations and it is important to ensurethat the test substance is available in the diet at the sameconcentration throughout the treatment period.9.4
37、 In addition to homogeneity and stability testing requiredby GLPs, it is recommended that concentrations of the testsubstance in the diet be confirmed by analysis at the beginningof the test.10. Test Organisms10.1 This practice is intended primarily to be used with theyoung of the following species:
38、 northern bobwhite (Colinusvirginianus), Japanese quail (Coturnix japonica), mallard(Anas platyrhynchos), and ring-necked pheasant (Phasianuscolchicus). Other species may be used, but changes in diet,caging, and other factors may be necessary (12, 13).10.2 If laboratory or commercially reared birds
39、are used inthis practice they must come from the same source, and be ofthe same age, because different strains or age cohorts canintroduce variability into the test. These birds should be similarin appearance to a wild species. The parentage and dietaryhistory of purchased birds should be known. If
40、captured wildbirds are used, they should come from the same source and beof similar maturity.10.3 Birds that are deformed, injured, emaciated, or pheno-typically different from normal birds must not be used as testanimals. The population of birds from which the test animals(treated and control) are
41、selected shall be considered unsuitablefor testing if mortality exceeds 5 % during the 3 days prior totesting.10.4 The preferred age for Japanese quail and northernbobwhite is 14 days; for ring-necked pheasants, 10 days; andfor mallards, 5 days (17). The preferred ages are based on theprobability th
42、at test birds of these ages will not survive for 5days without eating (see 12.1.4). Tests with younger or olderbirds also can be used to determine the LC50(14, 15, 16). Ifdata from one test are to be considered comparable with datafrom another test, the ages of birds between the two testsshould devi
43、ate no more than one or two days.10.5 Young birds of the species listed in 1.3 shall beconditioned to the test parameters of caging, food, water, andphotoperiod from the time they hatch or are acquired until theinitiation of the test. An acclimation period of at least 3 days isrequired (see 10.3). O
44、lder birds shall be conditioned for at least7 days.11. Procedure11.1 Range-Finding Test:11.1.1 To determine the test concentrations to be used in adefinitive test, a range-finding test may be conducted for 5 daysusing three to five widely spaced concentrations.11.1.2 One procedure is to use an initi
45、al concentration of atleast 5000 ppm with two to four geometrically spaced lowerconcentrations. If there is no mortality at the 5000-ppm level,and test procedures and numbers of birds per concentration arethe same as would be used in a definitive test, then therange-finding test may provide sufficie
46、nt information to negatethe need for a definitive test. If mortality does occur, thenrange-finding will suggest the approximate test concentrationsto be used in a definitive test.11.2 Definitive Test:11.2.1 Individual test birds should be randomly assigned togroups and to control and test diet conce
47、ntrations. Assignmentto groups and initial weighing of the test birds should be doneat the same time to avoid needless handling stress.11.2.2 Water, and treated or untreated diets, should beavailable ad libitum.11.2.3 The experimental (test and control) diets are avail-able for 5 days after which th
48、ey are replaced with untreatedfeed. Birds are held for a minimum of 3 days followingtreatment. In some situations, it may be necessary to extend theobservation period in order to investigate prolonged or delayedeffects.11.2.4 Body weight must be recorded at the initiation andconclusion of the treatm
49、ent and observation phases. Feedconsumption must be recorded for both the treatment andobservation phases; it is recommended that consumption dur-ing the treatment phase be recorded separately for the first twodays and the last three days. Additional information may begathered by measuring feed consumption daily. If the studycontinues beyond 8 days, body weight and feed consumptionshould be recorded weekly. Mortality, behavioral abnormali-ties, and other signs of toxicity should be recorded each dayduring the test.11.2.5 Photoperiod during the test should be the same asduring
