ASTM E653-91(1996)e1
(Test Method)Standard Method for Testing Effectiveness of Aerosol and Pressurized Space Spray Insecticides Against Flying Insects
Standard Method for Testing Effectiveness of Aerosol and Pressurized Space Spray Insecticides Against Flying Insects
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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e1
Designation: E 653 – 91 (Reapproved 1996)
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.
e NOTE—Editorial changes were made throughout in October 1996.
1. Scope less than 80 % of the particles have an arithmetic mean
diameter of 30 μm and many are 50 μm to 100 μm in mean
1.1 This test method determines the effectiveness of aerosol
diameter. Pressurized sprays shall be no less effective than the
and pressurized space-spray insecticides against house flies
selected reference standards when tested against house flies at
(Musca domestica L) and, with modifications in dosage, other
no more than twice the dosage specified for the selected
flying insects.
reference standard.
1.2 The test may be conducted using approximately 100
house flies per test (small group) or 500 flies per test (large
4. Summary of Test Method
group).
4.1 If the small-group method is used, ten tests are run on
1.3 This standard does not purport to address all of the
the Official Test Aerosol (using the selected reference stan-
safety concerns, if any, associated with its use. It is the
dard) and on each of the specimens in parallel. The specimens
responsibility of the user of this standard to establish appro-
of a series shall be randomized in the order of testing.
priate safety and health practices and determine the applica-
4.2 If the large-group method is used, the test is conducted
bility of regulatory limitations prior to use.
as in 4.1, with the exception that five, rather than ten tests are
2. Referenced Document required.
4.3 The average percentage mortality of the test insecticide
2.1 ASTM Standards:
compared with that of the selected reference standard is the
E 652 Test Method for Nonresidual Liquid Household In-
basis for assigning either Grade A (aerosol or pressurized space
secticides Against Flying Insects
spray) or Grade B (pressurized space spray) rating to the test
3. Terminology
specimen.
3.1 Definitions of Terms Specific to This Standard:
5. Significance and Use
3.1.1 aerosols—for this test method, the spray from aerosol
5.1 This test method provides a satisfactory means of
dispensers should be in finely divided form in which 80 % or
determining the relative efficacy of aerosol and pressurized
more of the individual spray particles have an arithmetic mean
space spray insecticide formulations against house flies (Musca
diameter of 30 μm or less, and none of the spray particles have
domestica, L) strains.
a diameter of more than 50 μm. Aerosols shall be no less
5.2 Test data obtained by this test method may also be
effective than the selected reference standards when tested
adequate to support label claims for the use of the product
against house flies at the same dosage or less.
against mosquitoes, gnats, flying moths, wasps, and certain
3.1.2 fly culture—all adults resulting from the seeding of
other small flying insects. This test method is not designed to
eggs collected at one time on a given date.
measure the residual activity.
3.1.3 knocked-down flies—all adult test flies incapable of
5.3 As a biological test, it is subject to the variations that
coordinated movement (moribund).
accompany the reaction of living organisms. It should be
3.1.4 pressurized sprays—these products deliver mist
employed under the supervision of personnel familiar with the
sprays intermediate between aerosols and sprays intended to
biological testing of insecticides.
deposit an insecticidal residue. They produce sprays in which
6. Apparatus
6.1 Reference Standard —The reference standard shall be
This test method is under the jurisdiction of ASTM Committee E-35 on
Pesticides and is the direct responsibility of Subcommittee E35.12 on Insect Control
one of the current selected reference standards from the
Agents. It was originally developed by the Chemical Specialties Manufacturers
Association (CSMA).
Current edition approved July 15, 1991. Published September 1991. Originally The Official Test Aerosol (Selected Reference Standard) has been found
published as E653 – 78. Last previous edition E653 – 84. suitable for this test and is available from CSMA, 1913 Eye Street N.W.,
Annual Book of ASTM Standards, Vol 11.05. Washington, DC 20006.
Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
E 653
container in which it is supplied. The selected reference device is used, it must produce gentle suction, have a suffi-
standards are (a) OTA-II to be used for oil-based aerosol ciently large receptacle to prevent crowding the flies, and be
products, or (b) TOAPS to be used for water-based aerosol cleaned after each test.
products. When reporting results, the selected reference stan- 6.10 Adult Fly Food—Dissolve 5 % of spray-dried (or
dard should be identified by its date. instant) nonfat dry milk solids and 2 % granulated sugar in
6.2 Test Specimen Dispenser—No restriction is placed on water. A40 % formalin solution may be added at the rate of
the test specimen dispenser. However, it should be noted that 1 + 1500 to delay spoiling.
the test results apply only to the test specimen as dispensed 6.11 Shallow Containers—Containers shall not be more
from the particular unit employed. than 0.75 in. (19 mm) high, to hold 5 % sugar solution as food
6.3 Fly Cages —Cages of any convenient type may be used for paralyzed flies. A gauze-wrapped ball of cotton saturated
3 3
if they provide at least 1 in. (16 cm ) of space per fly and have with sugar solution is also satisfactory.
at least two sides and the top screened. The cages should be 6.12 Larval Medium Containers, cylindrical glass battery
constructed of metal or other suitable material, and fitted with jars, approximately 6 in. (152 mm) in diameter and 9 in. (229
a sleeve opening, rubber membrane, or door. A detachable floor mm) high, or other suitable containers.
is preferable to facilitate cleaning and the insertion of a paper 6.13 Larval Medium—For each container, mix 340 g of
floor covering. CSMA Standard Fly Larval Medium with approximately 750
6.4 Rearing Room—A room of any convenient size, free of cm of an aqueous suspension containing 15 g of moist cake
strong drafts, and maintained at 80 6 2°F (27 6 1°C), with a yeast or5gof active dry yeast and 10 cm of nondiastatic
relative humidity of 50 6 5 %. The rearing room should be diamalt. Thoroughly mix this combination until a loose, fluffy
separate from the testing room and ventilated to minimize consistency is obtained, transfer it to the container without
odors and gases from fermenting media. packing, cover the container with a cloth or other suitable
6.5 Testing Room—A room of any convenient size capable cover, and set it in the rearing room. The amount of suspension
of holding the test chamber, with adequate additional space to required for best rearing results will need to be determined in
permit efficient performance of the tests. The room shall be each laboratory and may be varied to prevent mold growth. It
maintained at 80 6 2°F (27 6 1°C), with a relative humidity is suggested that the medium be prepared in the late afternoon
of 506 5%. of the day before egg collection.
6.6 Test Chamber—A standard Peet-Grady chamber meet- 6.14 Calibrated Centrifuge Tube, Pipet, Pit, or Cell,tobe
ing the general specifications given in Test Method E 652. If a used for the measurement of 2000 eggs (0.1 cm of settled eggs
larger chamber is used, it is recommended that its dimensions equals approximately 700 eggs).
approximate a normal size room. 6.15 Air-Separation Apparatus—An air-separation appara-
6.6.1 When a Peet-Grady chamber is used, the actuator tus, constructed according to the specifications of Goodhue and
nozzles should be directed so that the spray goes through a Linnard, will provide a rapid means of separating pupae from
port. the larval-rearing medium. The apparatus employs a suction
6.6.2 Adjustable fixtures may be used to hold the dispensers pipe, blower, and cyclone separator to remove dried vermicu-
and distribute the sprays from the same place and angle for lite (placed on the fly larval medium prior to pupation) from the
each test. Since different adjustments may be required for heavier pupae.
various test dispensers, the spray pattern from new dispensers 6.16 Vermiculite.
should be determined prior to testing. Successful use has been 6.17 Shallow Tray.
reported with a fixture adjusted to position the dispenser 8 in. 6.18 Clean Cloths.
(203 mm) from the ceiling and 10 in. (254 mm) from a corner 6.19 Ethyl Alcohol, Ethyl Alcohol Containing 10 % Ac-
of the Peet-Grady chamber. etone, Soap and Water, or Detergent and Water.
6.7 Exhaust Fan—An exhaust fan, capable of moving air 6.20 Oviposition Medium.
through the test chamber at not less than 1000 ft /min (0.5 6.21 Test Insect—The test insect shall be the adult house fly,
m /s), shall be used to ventilate the chamber after each test. It Musca domestica L, reared from the current official CSMA
shall be arranged with adequate piping to exhaust the chamber non-resistant house fly strain. Healthy test groups with an
vapors in a safe manner. average age of 4 days shall be used and individual flies in the
6.8 Paper—Unsized, nonglazed, absorbent paper (such as test groups shall not be less than 3 or more than 6 days old at
brown kraft or gray bogus) shall be used to cover the test the time of testing. The strain shall be of such susceptibility
chamber floor. Two overlapping sheets of 36 to 40 in. (0.9 to that the Official Test Insecticide (OTI) will cause a 24-h
1.0 m) in width or one sheet of 6 ft (1.8 m) in width may be
employed. No special weight is specified, but 60 to 80-lb (27
The CSMA Standard Fly Larval Medium is available from the Ralston Purina
to 36-kg) gray bogus has been found to be satisfactory.
Co., P.O. Box 337, Richmond, IN 47374.
6.9 Apparatus for Collecting Treated Flies—Any conve-
Yeasts and diamalt, manufactured by Standard Brands, Inc., are available from
nient means of picking up the paralyzed flies without injuring
local distributors.
Goodhue, L. D., and Linnard, C. E., “Air Separation Apparatus for Cleaning
or appreciably disturbing them may be used. If a vacuum
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.
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
E 653
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
7.1 Collect eggs for a period of not longer than 16 h from flies at all times until testing.
7.6 Hold the adult flies until they are about 4 days old (see
food dishes or other oviposition media in cages containing
mature flies not more than 8 days old. Fresh oviposition 6.21). They will then be ready for testing.
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) at 80 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
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