1、Designation: E 1517 99 (Reapproved 2006)Standard Test Method forDetermining the Effectiveness of Liquid, Gel, Cream, orShampoo Insecticides Against Human Louse Ova1This standard is issued under the fixed designation E 1517; 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 (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This test method determines the effectiveness of ovi-cidal mater
3、ials in liquid, gel, cream, or shampoo form againstthe ova (that is, eggs or nits) of the human louse, Pediculushumanus.1.2 This test method is intended for use by those wishing todevelop efficacy data or compare ovicidal formulations forhuman louse ova control.1.3 This test method consists of five
4、replicates for a statis-tical comparison of formulations.1.4 The values stated in SI units are to be regarded as thestandard. The values given in parentheses are for informationonly.1.5 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theres
5、ponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Terminology2.1 Definitions of Terms Specific to This Standard:2.1.1 hatchedthose eggs (nits) from which the nymph hasemerged compl
6、etely. An empty, clear egg case with theoperculum clearly open.2.1.2 unhatchedthose eggs that are opaque. The opercu-lum is closed or the nymph is partly emerged.3. Summary of Test Method3.1 Five replicates of 30 eggs are immersed in a testcompound for a set period of time, washed, rinsed, blotted,d
7、ried, and incubated.3.2 Five control replicates are attached to human hair andprocessed as the treatment replicates, but with immersion inwater.3.3 Percent egg mortality, corrected by Abbotts Formula, isdetermined.4. Significance and Use4.1 This test method is a standardized test for the gatheringof
8、 efficacy data for human louse ovicides.4.2 Data collection in this manner is suitable for productdevelopment and comparison, and for review by regulatoryagencies, to support the registration of human louse ovicidalproducts.5. Apparatus and Materials5.1 ApplicatorsEgg-infested hairs are attached to
9、the endof a wooden applicator stick with duct tape such that 30 nits areon 1 to 3 hairs. Each replicate of 30 eggs is examined under adissecting microscope to confirm viability. Any eggs that areshrunken or with other indications of being nonviable areexcluded.5.2 BeakersA 100-mL beaker is used to c
10、ontain 60 mL oftest ovicide and another to contain 60 mL of water (control),into which the applicators are dipped. A 1000-mL beaker isused for washing the eggs.5.3 Heating SurfaceA slide dryer that provides heat ofapproximately 32C (90F).5.4 Incubator, capable of maintaining a temperature of 31.76 0
11、.5C (89F) and a relative humidity of 60 6 10 %.5.5 Water Bath, capable of maintaining a temperature of32C (90F).5.6 Wash Bottle, Stop Watch, and Dissecting Scope.5.7 Test InsectThe human louse, Pediculus humanus.25.8 Positive Control TreatmentSixty mL of solutionknown to give 65 to 95 % mortality of
12、 louse eggs when usedunder these test conditions.6. Rearing of Test Insects6.1 The adult human lice are blood fed on the shaven bellyof a restrained rabbit.6.2 The lice are transferred to human hair cuttings, held ina petri dish, and incubated for 24 h for oviposition to occur.1This test method is u
13、nder the jurisdiction of ASTM Committee E35 onPesticides and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.22 on Pesticide Formulations and Delivery Systems.Current edition approved April 1, 2006. Published April 2006. Originallyapproved in 1993. Last previous editio
14、n approved in 2005 as E 1517 99 (2005).2A strain of the human body louse, Pediculus humanus, is maintained by InsectControl and Research, Inc., Baltimore, MD 21228-1199. The strain was establishedfrom a U.S. Department of Agriculture Gainesville colony. It is a susceptible strainand, through selecti
15、on, has been adapted to the New Zealand White rabbit.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.6.3 The lice are then allowed to crawl off the egg-infestedhairs by placing them on a rabbits belly, leaving the hairs withattached
16、eggs for exposure.7. Procedure7.1 Use five replicates of each test formulation and fivecontrol replicates.7.2 Prepare 5 cohorts of eggs for each treatment to be testedincluding the control treatments. Each cohort consists of 30eggs (one to three hair shaft(s) attached with duct tape to awood applica
17、tor stick.7.3 Heat the test samples to 32 6 1C in the waterbath.7.4 Insert the taped ends (hairs) of the applicator sticks intothe test samples for 10 min of immersion.7.5 Wash the eggs in 900 mL of 32C tap water for 1 min byvigorous up and down movement of the applicator sticks withthe hairs attach
18、ed.7.6 Rinse the eggs with water from the wash bottle for 1min.7.7 Blot excess water with paper toweling.7.8 Transfer the hair with attached eggs to labelled petridishes and incubate.7.9 Follow the same procedure for the control replications,except substitute tap water for the test solution.7.10 Whe
19、n all control eggs have hatched (after approxi-mately 12 days), examine all replicates under a dissectingmicroscope to determine the numbers hatching and failing tohatch. Failure to hatch is recorded as mortality. Categorize eggsfailing to hatch as follows:7.10.1 Early stage (no visible differentiat
20、ion of the embryowhen viewed under 303);7.10.2 Late stage (visible differentiation of embryo whenviewed under 30X, typically eye spot is visible); and7.10.3 Emergent (nymphal louse has opened operculum andbegun to emerge, but died before emerging completelypartof nymphs body still within egg shell).
21、8. Analysis of Data8.1 Calculate the percentage of control eggs failing to hatch;if this exceeds 15 % the results should be discarded and the testrepeated.8.2 Correct all counts of treated eggs failing to hatch byAbbotts Formula (corrected % killed = (% alive control - %alive treated) 3 100 % 4 % al
22、ive control).3Confirm that thecorrected mortality experienced by the positive controls is 65to 95 %; if it is not, the results should be discarded and the testrepeated.8.3 Mortality data will be analyzed by appropriate statisticalprocedures, such as, analysis of variance (ANOVA), followedby Students
23、-Newman Keuls test to separate the means.9. Precision and Bias9.1 No precision data are available for this test method;however, ASTM Committee E35 is interested in conducting aninterlaboratory test program and encourages interested partiesto contact the staff manager, Committee E35, ASTM Head-quarte
24、rs.10. Keywords10.1 effectiveness; human louse ovicides; insecticidesASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. Users of this standard are expressly advised that determination of the validity of an
25、y such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years andif not revised, either reapproved or withdrawn. Your comments are i
26、nvited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of theresponsible technical committee, which you may attend. If you feel that your comments have not receiv
27、ed a fair hearing you shouldmake your views known to the ASTM Committee on Standards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the aboveaddress or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.org (e-mail); or through the ASTM website(www.astm.org).3Finney, D., Probit Analysis, Cambridge University Press, Cambridge, England,1962, pp. 8892.E 1517 99 (2006)2
