Effectiveness of Liquid, Gel, Cream, or Shampoo Insecticides Against Adult Human Lice

Designation: E938/E938M − 12
Standard Test Method for

Effectiveness of Liquid, Gel, Cream, or Shampoo
Insecticides Against Adult Human Lice1
This standard is issued under the fixed designation E938/E938M; 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 (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3. Summary of Test Method

1.1 This test method determines the effectiveness of pedicu- 3.1 Five replicates of 25 lice shall be used for each test
licidal materials in liquid, gel, or cream form, against the adult concentration or any other variable tested. Five water control
human louse, Pediculus humanus humanus, the surrogate replicates will be used on each day of testing.
subspecies for the human head louse (P.h. capitis). (Only gels 3.2 Percent louse mortality, corrected by Abbott’s Formula,
or creams that liquefy at 32°C [90°F] can be tested). is determined.2
1.2 This test method is for the use of those wishing to
develop efficacy data on adult lice. 4. Significance and Use
1.3 This test method consists of five replicates for a statis- 4.1 This test method should provide a consistent approach
tical comparison of formulations. both in terms of test insects and test procedures for the
gathering of efficacy data for pediculicides.
1.4 The values stated in either SI units or inch-pound units
are to be regarded separately as standard. The values stated in 4.2 Data collection in this manner should be suitable for
each system may not be exact equivalents; therefore, each product development and comparison. In addition, it should be
system shall be used independently of the other. Combining suitable for review by regulatory agencies.
values from the two systems may result in non-conformance 5. Apparatus and Materials
with the standard.
5.1 Test Container—A 9-dram plastic vial, screened at the
1.5 This standard does not purport to address all of the
bottom with 20-mesh screen, shall be used as the dipping
safety concerns, if any, associated with its use. It is the
vessel. A plunger, made from a plastic rod, and a circular
responsibility of the user of this standard to establish appro-
screen fits inside the vial. Plastics used should be as chemically
priate safety and health practices and determine the applica-
unreactive as possible. Plastic vials are to be discarded after
bility of regulatory limitations prior to use.
each test.
2. Terminology 5.2 Beakers—A 100- to 500-mL beaker is used to contain
2.1 Definitions of Terms Specific to This Standard: the pediculicide into which the test container is dipped.
2.1.1 morbid—unable to move towards heat 1 h after A1000-mL beaker is used as the container in which the lice are
treatment: sickly, but not necessarily dying; may recover by 24 washed after treatment.
h. 5.3 Heating Surface—A slide warmer that provides heat of
2.1.2 moribund—unable to move towards heat (and there- approximately 37°C [98°F].
fore food) 24 h after treatment; dying. 5.4 Incubator, capable of maintaining a temperature of
31.7°C [89°F] and 60 % RH.
1
5.5 Petri dishes, 8.9 cm in diameter and 1.3 cm deep.
This test method is under the jurisdiction of ASTM Committee E35 on
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct 5.6 Waterbath, capable of maintaining 32°C [90°F].
responsibility of Subcommittee E35.12 on Insect Control Agents.
Current edition approved April 1, 2012. Published May 2012. Originally
approved in 1983. Last previous edition approved in 2011 as E938 – 05(2011) ´1. 2
Abbott, W. S., “A Method of Computing the Effectiveness of An Insecticide,”
DOI: 10.1520/E0938-12. Journal of Economic Entomology, Vol 18, 1925, pp. 265–267.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
E938/E938M − 12
5.7 Dark Cotton Corduroy, 4 by 4 cm. 7.3 Place the 9-dram vial in the 100-mL pediculicide beaker,
5.8 Paper Toweling, Stop Watch, Forceps or Spoon, and and keep the lice under the pediculicide for 10 min.
Wash Bottle. 7.4 Remove the test container and blot the bottom of the
5.9 Test Insect—The test insect is the human body louse, container to remove any remaining liquid.
Pediculus humanus humanus. The present strain was estab- 7.5 Place the 9-dram vial into the 1000-mL beaker contain-
lished from the USDA Gainesville strain.3 It is a susceptible ing tap water at 32°C and agitate the container. At the end of 1
strain and, through selection, has adapted to a rabbit host. min, remove container, and gently wash the lice in a stream of
5.10 Host Animal—New Zealand white rabbits. tap water (32°C) from the wash bottle for 1 min.
7.6 Blot excess water with paper toweling.
6. Rearing of Test Insects
7.7 Transfer the lice to a clean 4 by 4-cm patch of dark
6.1 Collect eggs twice a week. This can be done when the
corduroy cloth. Use forceps to remove any lice that remain in
corduroy patches are placed on the rabbit. The adult lice leave
the container. Place corduroy patch in a petri dish, then cover
the patches to feed. The patches are then removed from the
with an additional corduroy patch.
rabbit. Any lice that do remain on the patches should be
removed. 7.8 Place the petri dish with lice in an incubator maintained
at 31.7°C and 60 % RH.
6.2 Place the patch containing eggs in a plastic container (10
by 7 cm) with a screened lid, and note the date on the container. 7.9 Make the first observation 1 h post treatment, and
Place the container in an incubator that is maintained at 31.7°C replace the petri dish in the incubator.
and 60 % RH. 7.10 To make an observation, place the lice on top of one
6.3 The eggs will hatch in approximately 7 days and a blood patch, which is on top of the second patch, in a petri dish. The
meal should be provided to the newly-hatched nymphs on day petri dish is then placed on the slide warmer (37°C) or heating
7. pad. Lice not dead or morbid will move to the lower patch
within 5 min.
6.4 Provide blood meals six days a week. Allow the lice to
feed on the shaved abdomen of a restrained rabbit. The rabbit 7.11 For the controls, repeat all of the above procedures,
is placed on its back on the restraining rack for approximately substituting tap water for the pediculicide.
30 min. Collect the lice after feeding by moving the corduroy
patches back and forth gently over the shaved area of the 8. Analysis of Data
rabbit. Most of the lice will attach to the patch. Pick up any 8.1 Numbers of dead and moribund lice are summed to give
remaining lice with a forceps or a spoon. mortality at 24 h.
6.5 Lice used for testing are usually 17 6 1 day old (as 8.2 All mortality counts are corrected to mean corrected
determined from the date of the first blood meal). percent mortality using Abbott’s formula:
6.6 Keep adult lice, for egg laying purposes, approximately Corrected % killed 5 [ ~ % alive control 2 % alive treated! /
three weeks (from time of hatching) and then discard. % alive control] 3 1003
7. Procedure 8.3 Prior to statistical analysis, percent mortalities will be

transformed using Arcsine Tranformation Tables.4 Then an
7.1 Place 25 adult lice, mixed sexes, in the bottom of the Analysis of Variance (ANOVA) will be performed and the
9-dram test container. Insert the screened plunger to keep the means will be separated by a suitable statistical procedure.
lice from floating to the surface.
7.2 Place the pediculicide to be tested in a 100- to 500-mL 9. Precision and Bias
beaker and introduce the beaker into a waterbath maintained at 9.1 No precision data is available for this test method,
32°C. Allow the test formulation temperature to stabilize prior however, Committee E35 is interested in conducting an inter-
to testing. laboratory test program and encourages interested parties to
contact the staff manager, Committee E35, ASTM
3
The present strain of Pediculus humanus humanus is maintained by ICR, Inc., Headquarters.
Baltimore, MD 21228. If you are aware of alternative suppliers, please provide this
information to ASTM International Headquarters. Your comments will receive
careful consideration at a meeting of the responsible technical committee,1 which
4
you may attend. Box, G., Hunter, W., Hunter, S., Statistics for Experimenters, Wiley, 1978.
2
E938/E938M − 12
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Designation: E938/E938M − 12

Standard Test Method for

1. Scope 3. Summary of Test Method

7. Procedure 8.3 Prior to statistical analysis, percent mortalities will be

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