1、Designation: E 652 91 (Reapproved 2003)Standard Test Method forNonresidual Liquid Household Insecticides Against FlyingInsects1This standard is issued under the fixed designation E 652; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revisio
2、n, 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 covers the determination of the relativeefficiency of household and industrial-use, c
3、ontact insecticidesdissolved in base oils.1.2 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 regulator
4、y limitations prior to use.2. Terminology2.1 Definitions:2.1.1 culture, nall adult flies resulting from the seeding ofeggs collected at one time on a given date.2.1.2 knocked-downpertaining to all test flies incapable ofcoordinated movement (moribund).3. Summary of Test Method3.1 Two methods for eva
5、luating liquid household insecti-cides are permitted as follows:3.1.1 For the small group method,2a minimum of 10replicates of approximately 100 flies each are exposed to a totalof 12 cm3of test insecticide per replicate.3.1.2 For the large group procedure, use two separate flycultures, four randomi
6、zed tests with 500 flies per replicateusing 10 replicates.3.2 The difference in percentage mortality of the OfficialTest Insecticide (OTI) (see 8.2.1) and the test insecticide is thebasis for evaluating the efficacy of the test insecticide by thesmall and large group test methods.4. Significance and
7、 Use4.1 This test method provides a satisfactory means ofdetermining the relative efficacy of spray formulations againsthouse flies (Musca domestica, L).4.2 Test data obtained by this test method may also beadequate to support label claims for the use of the productagainst mosquitoes, gnats, flying
8、moths, wasps, and certainother small flying insects. This test method is not designed tomeasure the residual action of the spray formulation.4.3 As a biological test, it is subject to the variations thataccompany the reactions of living organisms. It should beemployed under the supervision of person
9、nel familiar with thebiological testing of insecticides.5. Apparatus5.1 CSMA Pesticide Atomizer, fitted with a No. 631 cut offand a glass reservoir.35.2 Rearing RoomA room of any convenient size, free ofstrong drafts, and maintained at 80 6 2F (27 6 1C) with arelative humidity of 50 6 5 %. This room
10、 must be separatefrom the testing room and ventilated to minimize odors.5.3 Testing Room, maintained at 80 6 2F (27 6 1C) anda relative humidity of 506 5 %. This room may be of anyconvenient size capable of holding the standard Peet-Gradychamber with adequate additional space to permit efficientperf
11、ormance of the test.5.4 Peet-Grady Test Chamber (see Annex A1.).5.5 Cylindrical Glass Battery Jars, 6 in. (150 mm) indiameter and 9 in. (230 mm) high, or other suitable containers,to be used as fly larval medium containers.5.6 Calibrated Pipet, or graduate with 0.1-cm3graduations.5.7 Electric Fan.5.
12、8 Air Separation Apparatus for Recovering Puparia, con-structed according to the specifications of Goodhue and Lin-nard.45.9 Fly Cages, providing at least 1 in.3(16.4 cm3) of spaceper fly with a minimum of two sides and the top screened.Cages shall be constructed of metal or other suitable materiala
13、nd fitted with a sleeve opening, rubber membrane, or a door.1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and is the direct responsibility of Subcommittee E 35.12 on InsectControl Agents.Current edition approved July 15, 1991. Published September 1991. Originallypubl
14、ished as E 652 78. Last previous edition E 652 84.2“Peet-Grady Method,” Official Method of the Chemical Specialties Manufac-turers Association for Evaluating Liquid Household Insecticides.3Available from Chemical Specialties Manufacturers Assn. (CSMA), 1913 EyeSt., N.W., Washington, DC 20006.4Goodhu
15、e, L. D., and Linnard, C. E., “Air Separation Apparatus for CleaningFly Pupae,” Journal of Economic Entomology, Vol 43, 1950, p. 228.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.A detachable floor is preferable to facilitate clean
16、ing andinsertion of a paper floor covering.56. Reagents and Materials6.1 Adult Fly Food5 % spray-dried (or instant) nonfatmilk solids and 2 % granulated sugar dissolved in water (40 %formalin solution may be added at the rate of 1 + 1500 to delayspoiling). Each cage requires 15 cm3of food per 100 fl
17、ies perday.6.2 Larval Medium340 g of CSMA Standard Fly Me-dium6added to 750 cm3of an aqueous suspension containing15 g of moist cake yeast7(or5gofdryyeast7) and 10 cm3ofnondiastatic Diamalt7per container (see 5.5). Some modifica-tions in liquid content may be needed to give maximum larvalproduction.
18、6.3 Puparial MediumAn added 2-in. (51-mm) layer ofvermiculite on the dry top surface of the fly larval medium.7. Test Specimen and Sample7.1 The test insect must be the adult house fly (Muscadomestica L) reared from the current CSMA official resistanthouse-fly strain.7.2 Adult house flies in test gr
19、oups must be between 3 and6 days of age at the time of testing.8. Calibration and Standardization8.1 Apparatus:8.1.1 AtomizerMaintain pressure at a constant 12.5 6 0.5psi (86.2 6 3.4 kPa) as measured by a gage of not more than30-psi (207-kPa) capacity or a manometer. Calibrate theatomizer at 80 6 2F
20、 (27 6 1C) to deliver 12 cm3of OTI in24 6 1s.8.1.2 Test Chamber ContaminationConsider chamberscontaminated and unsatisfactory for use when test flies (3 to 6days old) held in the chamber for a 12 to 16-h period with food,but without insecticide treatment, show mortalities greater than10 %, or when o
21、ver 10 % of the flies are paralyzed within 30min after liberation.8.2 Reference Standards:8.2.1 Current Offcial Test Insecticide (OTI).39. Procedure9.1 House Fly Rearing Technique:9.1.1 Larval MediumMix the larval medium (see 6.2)thoroughly until a loose, fluffy consistency is obtained, transferit t
22、o the battery jar (or other container) without packing, coverwith a suitable cover, and place in the insectary. The amount ofsuspension required for best rearing results will need to bedetermined in each laboratory and it may be varied to preventmold growth. It is suggested that the medium be prepar
23、ed inthe late afternoon of the day before egg collection.9.1.2 EggsCollect eggs for a period not longer than 16 hfrom food dishes or other oviposition medium in cagescontaining mature flies not more than 8 days old. It issuggested that fresh oviposition medium be placed in fly cagesin the late after
24、noon for egg collection early on the followingmorning. Measure and seed the collected eggs without delay.Wash all the eggs together in tap water at room temperatureand measure groups of 2000 as accurately as possible. Thismay be done by allowing the eggs to settle in a calibrated pipetor graduate (0
25、.1 cm3of settled eggs is approximately 700), orthe eggs can be filtered and measured in calibrated pits or cells.Use 10 cm3of tap water to measure and to scatter the eggs ina pit or trench 0.5 in. (13 mm) deep which is located in thecenter of the surface of the larval medium. Cover the eggs withloos
26、e medium and place the covered containers in the insectarywith at least 1.5-in. (38-mm) separation to permit free aircirculation. The maximum temperature in the jar (about 3 dayslater) must not exceed 130F (54.4C). Under normal condi-tions more than 85 % of the eggs should hatch within 36 h ofthe ti
27、me they are laid.9.1.3 PupaeApproximately 3 to 4 days after the eggshave been seeded, a 2-in. (51-mm) layer of vermiculite may beadded on the surface of the larval medium to aid in pupaerecovery.8Mature larvae migrate to the top portion of themedium or to the vermiculite layer, and normally all larv
28、ae willhave pupated about 9 days after seeding the eggs. When thisoccurs, the portion containing pupae may be removed, pouredinto a shallow tray, and air-dried at room temperature. Anelectric fan may be used to hasten drying. Then separate thepupae from the dry medium or the vermiculite. Handle gent
29、lyand as little as possible to avoid injury to the pupae. Anymethod that permits at least 90 % of the flies to emerge isconsidered satisfactory.9.1.3.1 Air-Separation ApparatusAn air-separation appa-ratus (see 5.8) is used by several laboratories for cleaningpupae and has been found to be more rapid
30、 than the indicatedtray method. The device employs a blower, a cyclone collector,and a suction pipe to separate the heavier pupae from a layer ofvermiculite placed on the surface of the fly larval medium.89.1.3.2 Combine all of the pupae maturing on a given dayinto one lot, mix, and measure into tes
31、t unit groups. Each groupis held in a shallow dish and placed in a cage that provides atleast 1 in.3(16.4 cm3) of space per pupae.9.1.4 AdultsIf the large group procedure is used, the testunit consists of approximately 500 pupae. If the small groupprocedure is used, more than 500 pupae are placed in
32、 stockcages and adult flies are sampled prior to testing. Under normalrearing conditions, obtain at least 80 adult flies for each 100eggs seeded. Daily supply each cage of adult flies with 15 cm3of adult fly food for each 100 flies and prepare so as to preventthe flies from drowning.9.2 Test Procedu
33、re:9.2.1 Before a fly spray test is started, the Peet-GradyChamber must be clean and have clean paper on the floor, allports and other openings must be closed, the temperature must5Cages available from American Biological Supply Co., 1330 Dillon HeightsAve., Baltimore, MD 21228, have been found suit
34、able for this purpose.6The CSMA Standard Fly Medium is a product of Ralston Purina Company, P.O.Box 337, Richmond, IN 47374, and has been found suitable for this purpose.7The yeasts and Diamalt are products of Standard Brands, Inc., and have beenfound suitable for this purpose.8Incho, H. H., “A Rapi
35、d Method for Obtaining Clean House Fly Pupae,” Journalof Economic Entomology, Vol 47, 1954, p. 938.E 652 91 (2003)2be 80 6 2F (27 6 1C), and all windows must be shadedequally. In both the large-group and small-group procedures,only flies that are capable of flying may be liberated into thePeet-Grady
36、 chamber. In the large-group procedure, all flies inone cage are used in a single test; but in the small-groupmethod, a sample of approximately 100 flies, 65, is used ineach test. Samples may be taken by liberating the flies directlyinto the chamber and continuing until about 10 % of the fliesremain
37、 in the stock cage to be discarded. The order of spraytreatments must be randomized.9.2.2 Immediately after liberation of the flies in the cham-ber, spray a total of 12 cm3of insecticide in equal quantitiesthrough each spray hole. Slowly oscillate the nozzle of theatomizer in a horizontal plane to a
38、void spraying walls andceilings and to effect uniform distribution of the spray. Com-plete this procedure within 1 min from the time the sprayingwas started and keep the chamber closed at a constanttemperature in the range of 80 6 2F (27 6 1C) for a total of10 min. At the end of this period, open th
39、e ports and ventilatethe chamber with the exhaust fan while the flies are collected.9.2.3 Pick up the knocked-down flies (see section 2.2) inany manner that will not appreciably disturb or harm them andimmediately transfer them to the clean cages (see 5.9). Theseflies may be counted when they are pi
40、cked up or later,depending upon which time is most convenient. During thesubsequent 24-h recovery period, maintain the cage underrearing room conditions of temperature and humidity andsupply the treated flies with an adequate quantity of 5 % sugarsolution arranged so they cannot drown in it. A gauze
41、-wrappedball of cotton saturated with the 5 % sugar solution is satisfac-tory.9.2.4 Count the unaffected “up” flies in the chamber at theend of the 10-min exposure period and discard.9.2.5 After a test is completed, remove all toxic residuesfrom the chamber. Renew the paper on the floor and thorough
42、lyclean the inside walls and ceiling. Wipe with a clean clothsaturated with alcohol containing 10 % acetone, or wash withsoap and water to remove most toxic residues. Special cleaningmay be required to remove insoluble toxic residues fromcertain chemical compounds. It is recommended that laborato-ri
43、es make a standard practice of taking periodic contaminationobservations, employing a normal test fly group (see 8.1.2).10. Report10.1 Assembling the DataCount and record the number ofunaffected flies at the end of the 10-min exposure period.Count the dead flies 16 to 24 h later, preferably by remov
44、ingthem from the recovery cage. Only flies that show no sign oflife upon being touched may be counted as dead. If knocked-down flies were counted as they were collected, the sum ofknocked-down and unaffected flies yields the total number offlies in the test. If knocked-down flies were not counted as
45、collected, the recovered flies may be killed by a suitablemethod, after which they are counted. The sum of recoveredand dead flies yields the knocked-down flies, and this sumadded to the unaffected flies yields the total number of fliesused in the test. The mortality is the percent dead of total fli
46、esand the knockdown is the percent knocked-down of total flies.10.2 Test Format:10.2.1 Small Group ProcedureRun ten parallel tests onthe OTI and on each of the unknowns. Test each spray the samenumber of times on flies of the same culture (see 2.1) and testall sprays the same number of times on any
47、one day, with theunknown samples and the OTI randomized as to order oftesting. After the mortality data are obtained, calculate theaverage percent kills and determine the differences between theunknowns and the OTI.10.2.2 Large Group ProcedureThis evaluation is basedon differences in mortality deter
48、mined by a minimum of fourrandomized tests of the OTI and the unknowns. Select at leasttwo cultures of flies, using 500 flies per replicate in the fourrandomized tests.10.3 Evaluation:10.3.1 Conduct the tests in accordance with the procedurepreviously described.10.3.2 Use at least two cultures (see
49、2.1) of flies in makingan official evaluation.10.3.3 Do not use cages showing a combined mortality andcrippling greater than 15 % on the day of the test.10.3.4 An unknown insecticide must have a 10-min knock-down percentage equal to that of the OTI with a toleranceof 2. The kill by the OTI shall fall between 30 % and 55 % inall tests.11. Precision and Bias11.1 No statement is made about either the precision or thebias of Test Method E 652 for measuring the efficacy of theinsecticide spray formulation since the result merely stateswhether there is conform
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