ASTM_E_1517_-_13.pdf

Designation:E151713Standard Test Method forDetermining the Effectiveness of Liquid,Gel,Cream,orShampoo Insecticides Against Human Louse Ova1This standard is issued under the fixed designation E1517;the number immediately following the designation indicates the year oforiginal 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 test method determines the effectiveness of ovici-dal materials in liquid,gel,cream,or shampoo form against theova(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 replicates for a statis-tical comparison of formulations.1.4 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard,except for temperature measurements in Section 5.1.5 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 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 completely;an empty,clear egg case with theoperculum clearly open.2.1.2 unhatchedthose eggs that are opaque;the operculumis 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 dry,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 byAbbotts Formula,isdetermined.4.Significance and Use4.1 This test method is a standardized test for the gatheringof 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 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 contain 60 mL oftest ovicide and another to contain 60 mL of water(control),into which the applicators are submerged.A1000-mLbeaker 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 0.5C(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 Treatment(Optional)60 mL of solu-tion known to give 65 to 95%mortality of louse eggs whenused under these test conditions.1This test method is under the jurisdiction of ASTM Committee E35 onPesticides,Antimicrobials,and Alternative Control Agents and is the directresponsibility of Subcommittee E35.12 on Insect Control Agents.Current edition approved April 1,2013.Published April 2013.Originallyapproved in 1993.Last previous edition approved in 2006 as E1517 99(2006).DOI:10.1520/E1517-13.2A 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 selection,has been adapted to the New Zealand White rabbit.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 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.6.3 The lice are then allowed to crawl off the egg-infestedhairs by placing them on a rabbits belly,leaving the hairs withattached 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 applicator 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 mLof 32C tap water for 1 min byvigorous up and dow