ASTM E653-91(2003)
(Test Method)Standard Test Method for Effectiveness of Aerosol and Pressurized Space Spray Insecticides Against Flying Insects
Standard Test Method for Effectiveness of Aerosol and Pressurized Space Spray Insecticides Against Flying Insects
SIGNIFICANCE AND USE
This test method provides a satisfactory means of determining the relative efficacy of aerosol and pressurized space spray insecticide formulations against house flies (Musca domestica, L) strains.
Test data obtained by this test method may also be adequate to support label claims for the use of the product against mosquitoes, gnats, flying moths, wasps, and certain other small flying insects. This test method is not designed to measure the residual activity.
As a biological test, it is subject to the variations that accompany the reaction of living organisms. It should be employed under the supervision of personnel familiar with the biological testing of insecticides.
SCOPE
1.1 This method determines the effectiveness of aerosol and pressurized space-spray insecticides against house flies ( Musca domestica L ) and, with modifications in dosage, other flying insects.
1.2 The test may be conducted using approximately 100 house flies per test (small group) or 500 flies per test (large group).
1.3 This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation:E653–91(Reapproved 2003)
Standard Test Method for
Effectiveness of Aerosol and Pressurized Space Spray
Insecticides Against Flying Insects
This standard is issued under the fixed designation E 653; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.4 pressurized sprays—these products deliver mist
sprays intermediate between aerosols and sprays intended to
1.1 This test method determines the effectiveness of aerosol
deposit an insecticidal residue. They produce sprays in which
and pressurized space-spray insecticides against house flies
less than 80 % of the particles have an arithmetic mean
(Musca domestica L) and, with modifications in dosage, other
diameter of 30 µm and many are 50 µm to 100 µm in mean
flying insects.
diameter. Pressurized sprays shall be no less effective than the
1.2 The test may be conducted using approximately 100
selected reference standards when tested against house flies at
house flies per test (small group) or 500 flies per test (large
no more than twice the dosage specified for the selected
group).
reference standard.
1.3 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
4. Summary of Test Method
responsibility of the user of this standard to establish appro-
4.1 If the small-group method is used, ten tests are run on
priate safety and health practices and determine the applica-
the Official Test Aerosol (using the selected reference stan-
bility of regulatory limitations prior to use.
dard) and on each of the specimens in parallel.The specimens
2. Referenced Document of a series shall be randomized in the order of testing.
4.2 If the large-group method is used, the test is conducted
2.1 ASTM Standards:
as in 4.1, with the exception that five, rather than ten tests are
E 652 Test Method for Nonresidual Liquid Household In-
required.
secticides Against Flying Insects
4.3 The average percentage mortality of the test insecticide
3. Terminology
compared with that of the selected reference standard is the
basisforassigningeitherGradeA(aerosolorpressurizedspace
3.1 Definitions of Terms Specific to This Standard:
spray) or Grade B (pressurized space spray) rating to the test
3.1.1 aerosols—for this test method, the spray from aerosol
specimen.
dispensers should be in finely divided form in which 80 % or
more of the individual spray particles have an arithmetic mean
5. Significance and Use
diameter of 30 µm or less, and none of the spray particles have
5.1 This test method provides a satisfactory means of
a diameter of more than 50 µm. Aerosols shall be no less
determining the relative efficacy of aerosol and pressurized
effective than the selected reference standards when tested
spacesprayinsecticideformulationsagainsthouseflies(Musca
against house flies at the same dosage or less.
domestica, L) strains.
3.1.2 fly culture—all adults resulting from the seeding of
5.2 Test data obtained by this test method may also be
eggs collected at one time on a given date.
adequate to support label claims for the use of the product
3.1.3 knocked-down flies—all adult test flies incapable of
against mosquitoes, gnats, flying moths, wasps, and certain
coordinated movement (moribund).
other small flying insects. This test method is not designed to
measure the residual activity.
This test method is under the jurisdiction of ASTM Committee E35 on
5.3 As a biological test, it is subject to the variations that
Pesticides and is the direct responsibility of Subcommittee E35.12 on Insect Control
accompany the reaction of living organisms. It should be
Agents. It was originally developed by the Chemical Specialties Manufacturers
employed under the supervision of personnel familiar with the
Association (CSMA).
Current edition approved July 15, 1991. Published September 1991. Originally
biological testing of insecticides.
approved in 1978. Last previous edition approved in 1984 as E 653–84.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM The Official Test Aerosol (Selected Reference Standard) has been found
Standards volume information, refer to the standard’s Document Summary page on suitable for this test and is available from CSMA, 1913 Eye Street N.W.,
the ASTM website. Washington, DC 20006.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E653–91 (2003)
6. Apparatus 6.9 Apparatus for Collecting Treated Flies—Any conve-
nient means of picking up the paralyzed flies without injuring
6.1 Reference Standard —The reference standard shall be
or appreciably disturbing them may be used. If a vacuum
one of the current selected reference standards from the
device is used, it must produce gentle suction, have a suffi-
container in which it is supplied. The selected reference
ciently large receptacle to prevent crowding the flies, and be
standards are (a) OTA-II to be used for oil-based aerosol
cleaned after each test.
products, or (b) TOAPS to be used for water-based aerosol
6.10 Adult Fly Food—Dissolve 5 % of spray-dried (or
products. When reporting results, the selected reference stan-
instant) nonfat dry milk solids and 2 % granulated sugar in
dard should be identified by its date.
water. A40 % formalin solution may be added at the rate of
6.2 Test Specimen Dispenser—No restriction is placed on
1 + 1500 to delay spoiling.
the test specimen dispenser. However, it should be noted that
6.11 Shallow Containers—Containers shall not be more
the test results apply only to the test specimen as dispensed
than 0.75 in. (19 mm) high, to hold 5 % sugar solution as food
from the particular unit employed.
for paralyzed flies. A gauze-wrapped ball of cotton saturated
6.3 Fly Cages —Cages of any convenient type may be used
3 3
with sugar solution is also satisfactory.
if they provide at least 1 in. (16 cm ) of space per fly and have
6.12 Larval Medium Containers, cylindrical glass battery
at least two sides and the top screened. The cages should be
jars, approximately 6 in. (152 mm) in diameter and 9 in. (229
constructed of metal or other suitable material, and fitted with
mm) high, or other suitable containers.
asleeveopening,rubbermembrane,ordoor.Adetachablefloor
6.13 Larval Medium—For each container, mix 340 g of
is preferable to facilitate cleaning and the insertion of a paper
CSMA Standard Fly Larval Medium with approximately 750
floor covering.
cm of an aqueous suspension containing 15 g of moist cake
6.4 Rearing Room—Aroom of any convenient size, free of
6 3
yeast or5gof active dry yeast and 10 cm of nondiastatic
strong drafts, and maintained at 80 6 2°F (27 6 1°C), with a
diamalt. Thoroughly mix this combination until a loose, fluffy
relative humidity of 50 6 5 %. The rearing room should be
consistency is obtained, transfer it to the container without
separate from the testing room and ventilated to minimize
packing, cover the container with a cloth or other suitable
odors and gases from fermenting media.
cover, and set it in the rearing room.The amount of suspension
6.5 Testing Room—A room of any convenient size capable
required for best rearing results will need to be determined in
of holding the test chamber, with adequate additional space to
each laboratory and may be varied to prevent mold growth. It
permit efficient performance of the tests. The room shall be
is suggested that the medium be prepared in the late afternoon
maintained at 80 6 2°F (27 6 1°C), with a relative humidity
of the day before egg collection.
of 5065%.
6.14 Calibrated Centrifuge Tube, Pipet, Pit, or Cell,tobe
6.6 Test Chamber—A standard Peet-Grady chamber meet-
usedforthemeasurementof2000eggs(0.1cm ofsettledeggs
ing the general specifications given in Test Method E 652.Ifa
equals approximately 700 eggs).
larger chamber is used, it is recommended that its dimensions
6.15 Air-Separation Apparatus—An air-separation appara-
approximate a normal size room.
tus,constructedaccordingtothespecificationsofGoodhueand
6.6.1 When a Peet-Grady chamber is used, the actuator
Linnard, will provide a rapid means of separating pupae from
nozzles should be directed so that the spray goes through a
the larval-rearing medium. The apparatus employs a suction
port.
pipe, blower, and cyclone separator to remove dried vermicu-
6.6.2 Adjustable fixtures may be used to hold the dispensers
lite(placedontheflylarvalmediumpriortopupation)fromthe
and distribute the sprays from the same place and angle for
heavier pupae.
each test. Since different adjustments may be required for
6.16 Vermiculite.
various test dispensers, the spray pattern from new dispensers
6.17 Shallow Tray.
should be determined prior to testing. Successful use has been
6.18 Clean Cloths.
reported with a fixture adjusted to position the dispenser 8 in.
6.19 Ethyl Alcohol, Ethyl Alcohol Containing 10 % Ac-
(203 mm) from the ceiling and 10 in. (254 mm) from a corner
etone, Soap and Water, or Detergent and Water.
of the Peet-Grady chamber.
6.20 Oviposition Medium.
6.7 Exhaust Fan—An exhaust fan, capable of moving air
6.21 Test Insect—Thetestinsectshallbetheadulthousefly,
through the test chamber at not less than 1000 ft /min (0.5
Musca domestica L, reared from the current official CSMA
m /s), shall be used to ventilate the chamber after each test. It
non-resistant house fly strain. Healthy test groups with an
shall be arranged with adequate piping to exhaust the chamber
average age of 4 days shall be used and individual flies in the
vapors in a safe manner.
6.8 Paper—Unsized, nonglazed, absorbent paper (such as
brown kraft or gray bogus) shall be used to cover the test
The CSMA Standard Fly Larval Medium is available from the Ralston Purina
chamber floor. Two overlapping sheets of 36 to 40 in. (0.9 to
Co., P.O. Box 337, Richmond, IN 47374.
1.0 m) in width or one sheet of 6 ft (1.8 m) in width may be
Yeasts and diamalt, manufactured by Standard Brands, Inc., are available from
local distributors.
employed. No special weight is specified, but 60 to 80-lb (27
Goodhue, L. D., and Linnard, C. E., “Air Separation Apparatus for Cleaning
to 36-kg) gray bogus has been found to be satisfactory.
Fly Pupae,” Journal of Economic Entomology, Vol 43, 1950, p. 228.
Terra Lite Brand Vermiculite Soil Conditioner (No. 2 grade), available from
most garden or farm supply stores, has been found to be satisfactory for use in this
Cages available from American Biological Supply Co., 1330 Dillon Heights method.
Ave., Baltimore, MD 21228, have been found satisfactory for this method. Available from CSMA, 1913 Eye Street N.W., Washington, DC 20006.
E653–91 (2003)
test groups shall not be less than 3 or more than 6 days old at group in a shallow container and place in a cage with at least
3 3
the time of testing. The strain shall be of such susceptibility 1 in. (16 cm ) of space per pupae (see 6.3). If the large-group
that the Official Test Insecticide (OTI) will cause a 24-h method is used, each test group shall consist of approximately
mortality of 30 to 55 %, with approximately 95 % of the flies 500 pupae. If the small-group method is used, more than 500
paralyzed within 10 min following the spray application in pupae are placed in stock cages, and adult flies are sampled
accordance with Method E 652. At least two cultures of flies prior to testing. Under normal rearing conditions, at least 80
meeting these specifications shall be used in making an official adult flies should be obtained from each 100 eggs seeded.
evaluation. Daily supply each cage of flies with 15 cm of adult fly food
(see 6.10) for each 100 flies, and prepare so as to prevent the
7. Rearing of Test Insects
flies from drowning. Satisfactory food shall be available to the
flies at all times until testing.
7.1 Collect eggs for a period of not longer than 16 h from
food dishes or other oviposition media in cages containing 7.6 Hold the adult flies until they are about 4 days old (see
6.21). They will then be ready for testing.
mature flies not more than 8 days old. Fresh oviposition
medium may be placed in the cages in the late afternoon, and
8. Preparation of Apparatus
the eggs collected early the following morning.
8.1 Reference Standards (OTA II and TOAPS) and Test
7.2 Measure and seed the collected eggs without delay, as
Specimen Dispensers—Prior to use, calibrate the selected
follows:
reference standards and test aerosols or pressurized space
7.2.1 Wash the eggs in tap water at room temperature and
sprays (see 6.2)at80 6 2°F (27 6 1°C) to determine the rate
measure groups of 2000 as accurately as possible. This may be
of delivery in grams per second.
done by allowing the eggs to settle in a calibrated pipet or
8.2 Test Chamber—Before a test is started, clean the Peet-
centrifuge tube containing tap water, or the eggs can be filtered
Grady chamber, place clean paper on the floor, close all ports
and measured in a calibrated pit or cell.
and other openings, maintain the temperature at 80 6 2°F (27
7.2.2 Use 10 cm of tap water to measure and scatter the
6 1°C), and, equally shade all windows.
eggs in a pit or trench, 0.5 in. (13 mm) deep in the center of the
8.2.1 Chambers are considered to be contaminated and
larval medium (see 6.13).
unsatisfactory for test purposes when test flies held for a 12 to
7.2.3 Cover the eggs with loose medium, replace the con-
16-h period with food, but without insecticide treatment, show
tainer covers, and place the containers in the rearing room at
mortalities in excess of 10 %; or when more than 10 % of the
least 1.5 in. (38 mm) apart to permit free air circulation.
flies are knocked-down within 30 min after liberation into the
7.2.4 The maximum temperature in the larval medium
test chamber.
(about 3 days later) shall not exceed 130°F (54°C). Under
8.2.2 Periodic contamination observations, employing a
normal conditions, more than 85 % of the eggs should hatch
normal fly test group, should be a standard practice.
within 36 h of the time they were laid.
7.3 Mature larvae migrate to the top portion of the rearing
9. Procedure
medium or into a vermiculite layer, and normally all will have
9.1 In both the small and large group procedures, liberate
pupated by about the ninth day after seeding the eggs. When
into the test chamber only those test insects that are capable of
this occurs, the portion containing pupae is removed, poured
flying. Do not use cages showing a combined mor
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