ASTM E565-95

Test Method for Efficacy of a Single-Dose Acute Rodenticide Under Laboratory Conditions for Commensal Rodents (Withdrawn 2000)

SCOPE
1.1 This test method measures the efficacy of ready-to-use dry bait formulations prepared with single-dose acute rodenticides for the control of commensal rodents, including the Norway rat (Rattus norvegicus), roof rat (R. rattus), and the house mouse (Mus Musculus). This test method may also be applicable to some other species of rodents having similar behavioral patterns, physiology, and feeding preferences.
1.2 This test method is for use in developing efficacy data.
1.3 This test method also provides manufacturers, formulators, and other with a test procedure for monitoring product quality.
1.4 This test method, within limits, enables users of large quantities of commercial rodenticides to evaluate the efficacy of specific lots, or compare the efficacy of different formulations and different toxicants, or both.
1.5 This test method reduces many variables, thereby permitting replication or duplication of tests with reasonable accuracy.
1.6 This test method is not intended to be so restrictive that it will inhibit incentives towards the development of safer or more effective compounds or innovative approaches to bait formulations.
1.6.1 When justified by sound biological data or logical conclusions based on sound data, reasonable variations to some items in the test protocol may be made.
1.7 To ensure the quality and reliablity of data developed using this test method, good laboratory practices should be followed (see 4.1).
1.8 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for infomation only.
1.9 This test method represents the combined, several-year efforts of more than 70 scientists, users, and laboratory specialists from several countries and has not been superseded by better documentation (1995); therefore, it is the view of the committee that it be continued for the reference benefit fo any new concerns in the United States and abroad.
1.10 This standard does not purport to address all of the safety concerns, 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.

General Information

• Status: Withdrawn
• Publication Date: 09-Nov-1995
• Withdrawal Date: 09-Oct-2000
• ICS: 71.100.99 - Other products of the chemical industry
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: E 565 – 95
Standard Test Method for
Efficacy of a Single-Dose Acute Rodenticide Under
Laboratory Conditions for Commensal Rodents
This standard is issued under the fixed designation E 565; 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 1.9.1 Attention is directed to related ASTM documents, Test
Methods E 1163 and E 1372, that deal with similar situations;
1.1 This test method measures the efficacy of ready-to-use
however, were developed for different purposes.
dry bait formulations prepared with single-dose acute rodenti-
1.10 This standard does not purport to address all of the
cides for the control of commensal rodents, including the
safety concerns, if any, associated with its use. It is the
Norway rat (Rattus norvegicus), roof rat (R. rattus), and the
responsibility of the user of this standard to establish appro-
house mouse (Mus musculus). This test method may also be
priate safety and health practices and determine the applica-
applicable to some other species of rodents having similar
bility of regulatory limitations prior to use.
behavioral patterns, physiology, and feeding preferences.
1.2 This test method is for use in developing efficacy data.
2. Referenced Documents
1.3 This test method also provides manufacturers, formula-
2.1 ASTM Standards:
tors, and other with a test procedure for monitoring product
E 1163 Test Method for Estimating Acute Oral Toxicity in
quality.
Rats
1.4 This test method, within limits, enables users of large
E 1372 Test Method for Conducting A90-Day Oral Toxicity
quantities of commercial rodenticides to evaluate the efficacy
Study in Rats
of specific lots, or compare the efficacy of different formula-
tions and different toxicants, or both.
3. Summary of Test Method
1.5 This test method reduces many variables, thereby per-
3.1 This test method is based upon the principle of a
mitting replication or duplication of tests with reasonable
free-feeding choice between a toxic bait formulation and a
accuracy.
nontoxic, challenge diet offered to a selected group of test
1.6 This test method is not intended to be so restrictive that
rodents.
it will inhibit incentives towards the development of safer or
3.2 The test animals are offered the toxic bait formulation
more effective compounds or innovative approaches to bait
and challenge diet formulation for a specified period of time
formulation.
and the efficacy is measured by the percent mortality achieved.
1.6.1 When justified by sound biological data or logical
conclusions based on sound data, reasonable variations to some
4. Animal Facilities
items in the test protocol may be made.
4.1 No precise physical requirements for animal accomoda-
1.7 To ensure the quality and reliability of data developed
tions are set forth. However, the animal facility shall (a) meet
using this test method, good laboratory practices should be
the established guidelines suggested by the Institute of Labo-
followed (see 4.1).
ratory Animal Resources, or (b) be approved by such organi-
1.8 The values stated in SI units are to be regarded as the
zations as the American Association of Accreditation of Labo-
standard. The values given in parentheses are for information
ratory Animal Care (AAALAC).
only.
4.2 Maintain temperatures within the range from 18 to 29°C
1.9 This test method represents the combined, several-year
(65 to 85°F) and record daily using a maximum-minimum
efforts of more than 70 scientist, users, and laboratory special-
thermometer, hygrothermograph, or other suitable equipment.
ists from several countries and has not been superseded by
4.3 Strong air currents from heaters or air conditions shall
better documentation (1995); therefore, it is the view of the
not blow directly onto test or reference animals.
committee that it be continued for the reference benefit of any
4.4 The normal recommended range of humidity for labo-
new concerns in the United States or abroad.
ratory rats and mice is 40 to 70 %.
4.5 Natural lighting (through windows) is adequate. How-
ever, artificial lighting is acceptable and may be preferred when
This test method is under the jurisdiction of ASTM Committee E-35 on
controlled by time clocks. Artificial lighting may be correlated
Pesticides and is the direct responsibility of Subcommittee E35.17 on Verebrate
Control Agents.
Current edition approved Nov. 10, 1995. Published January 1996. Originally
published as E 565 – 76. Last previous edition E 565 – 90. Annual Book of ASTM Standards, Vol 11.05.
Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.
E 565
reasonably well to the natural day and night periods where the cially available strain of albino mice is acceptable.
tests are being conducted, or may be set to provide a light
7. Source of Test Animals
regime anywhere from 12 to 16 h is recommended (see 1.6.1).
7.1 For each test, laboratory rats or mice may be reared by
4.6 Artificial lighting may be of the incandescent or fluo-
the testing laboratory or purchased from the same commercial
rescent types.
supplier at the same time (as one lot).
4.7 The total reversing of the natural photoperiods of the
7.2 In all instances, the test rodents shall be selected on a
test animals by timed lighting is not recommended (see 1.6.1).
random basis within the scope of the other limitations placed
4.8 Light intensity is not specified. However, light intensity
on the condition of the test animals described herein (see
shall be relatively uniform for all test cages, whether natural or
Section 8).
artificial lighting is used. In some instances, natural lighting
7.3 Wild stock of Norway rats, roof rats, or house mice may
may have to be supplemented with artificial lighting to achieve
be live-trapped from the wild, reared in outdoor colonies, or
this. Light levels should not exceed 323 13 (30 ft-candles)
reared under laboratory conditions that will permit them to
about 1.0 m (3.3 ft) above the floor. This will reduce retinal
retain much of their natural physiological and behavioral
lesions in albino rats and should be sufficient for performance
characteristics. Breeding stock used for rearing wild rodents
of routine animal care.
shall not be selected for docile qualities or other characteristics
4.9 Animals on test shall not be subjected to undue or
that significantly alter their natural wild tendencies.
unnecessary stress from noise or human activities (that is,
movement). Human activity within the animal test room shall
8. Condition of Animals
be restricted to performing test protocols (see 13.7.1).
8.1 All animals shall be in apparent good health with no
obvious external open wounds, scars, or evidence of rhinitis.
5. Housing
8.2 Females should not be pregnant (see 9.3).
5.1 The test and reference rats or mice shall be maintained
8.3 Test and reference animals shall not knowingly have
in all metal-cages designed to hold laboratory rats or mice.
been previously exposed to any pesticide (such as rodenticide)
5.1.1 Cages shall have a minimum floor space of 387
except as follows:
2 2 2 2
cm (60 in. ) for rats and 258 cm (40 in. ) for mice. Maximum
8.3.1 Animals previously anesthetized or medicated either
cage sizes shall be no more than three times the minimum floor
orally or by injection shall not be used for test purposes within
space.
the 7-day period following the last treatment.
5.2 Preferably, these cages should be suspended in racks.
8.3.2 Ectoparasite control with appropriate concentrations
5.3 Cages shall have a wire mesh or screen bottom to permit
of sevin, malathion, or pyrethrin dusts is not considered
feces and urine to pass through.
medication and shall be applied externally to both test and
5.4 A tray or other arrangement under the cages shall be
reference wild animals on Day 1 of confinement.
used to permit recover of spilled foods. Each spilled food must
9. Pretest Conditioning
be separately weighed and these weights shall be added to the
unconsumed bait or challenge diet to determine consumption.
9.1 Test and reference animals shall be caged individually in
5.4.1 Damp spilled food shall be dried to approximately its
cages of the same type or kind that are to be used in the test.
original moisture content before weighing.
9.2 Laboratory strains of rodents obtained from a commer-
5.5 Water shall be available ad libitum. Glass water bottles
cial supplier or from other sources shall be conditioned
equipped with stainless steel or glass sipper tubes are recom-
(acclimated) to the test laboratory for a minimum of 3 days
mended. Gravity-fed automatic or open-cut waterers are not
before the start of a test regime.
recommended.
9.3 Wild-captured rodents or wild species from mixed sex
colonies may be separated by sex and held caged in groups or
6. Test Animals
individually for a period of time exceeding the gestation period
6.1 Ready-to-use bait formulations containing single-dose
to ensure that no females are pregnant.
acute toxicants shall be tested on all species for which they are
9.4 As a minimum, the last 7 days of the holding period for
to be used.
wild rodents shall be under laboratory conditions (that is,
6.2 Laboratory (domesticated) strains of albino rats and
temperature, humidity, and lighting) comparable to those of the
mice may be valuable for preliminary screening or for finding
testing animal room, if not actually in the testing room.
the range of appropriate toxicant concentrations, bait materials,
9.5 All rodents shall receive a nutritionally balanced com-
and other useful additives, but they cannot be substituted for
mercial rodent diet and water ad libitum during the pretest
the wild strains in the final test.
period.
6.3 Wild rats and mice shall be used in the final testing of a
9.6 Food consumption shall be measured and recorded
bait (see 7.3).
during this period. Water consumption could be monitored, if
6.3.1 Some correlation established between the laboratory
desired. Rodents failing to feed or drink normally should be
and the wild rodents of the same species may prove helpful for
removed from the test or reference groups, or both.
future tests with a particular bait formulation or rodenticide.
10. Number of Test Animals
6.4 Where laboratory rats are used, Wistar or Sprague-
Dawley derived albino or Long Evans (hooded) rats have been 10.1 Twenty test rodents (10 male, 10 female) shall be the
selected as the strains of choice for the sake of uniformity. minimum number to be used for conducting a single acceptable
6.5 Where laboratory mice are used an recognized commer- laboratory efficacy test.
E 565
TABLE 1 Weight of Test Animals
10.2 Although optional, additional laboratory tests may be
conducted to provide a more statistically sound base of Weight (g)
efficacy.
Maximum Acceptable
Test Animal
Min Max Difference in Average Weights
10.3 Ten rodents may prove adequate for manufacturers or
Between Sexes (12.4)
formulators monitoring for quality control or for range-finding
Laboratory mice 15 35 5
in screening experimental compounds.
Wild house mice 10 25 3
10.4 A reference group of twenty animals is recommended
Laboratory rats 150 300 50
for each test group (see 11.2). Reference groups of only 10 Wild Norway rats 150 400 65
Wild roof rats 100 200 33
animals may be acceptable for certain tests (see 10.3).
10.5 The reference group(s) may be used as a test group in
subsequent tests, provided they have not been exposed to the
the females and males for each test has been provided in Table
challenge diet and they meet initial specified criteria. 1.
13. Procedure
11. Reference Animals
13.1 Using conditioned (acclimated) animals (see 9.2 and
11.1 The term “reference animals,” is used to denote a
9.4) house 20 animals (10 males, 10 females) in individual
group of animals maintained very similarly to the test animals
cages. Record the weight to at least the nearest 1 g for each
for the purpose of determining mortality due to illness, injury
animal (see 12.1).
in handling or shipment, or other factors unrelated to the
13.2 Number the animal cages.
toxicant. Reference animals are not maintained in sufficiently
13.3 Equip each test cage with two identical food contain-
comparable conditions to the test animals to be considered
ers, one for the toxic bait and the other for the challenge diet,
control animals in the true sense of the definition.
to provide each rodent with a free-choice feeding situation.
11.2 A reference group of twenty animals is recommended
13.3.1 Procedures for “Group Choice Test Method.”
for each test group. Reference groups of only 10 animals may
13.3.1.1 Include here test design that uses group caging of
be acceptable for certain tests and shall be maintained to
animals. Also, add reference group in 11.3 to be consistent.
monitor mortality that might occur in a group not receiving the
13.4 Position the food containers on the right and left sides
toxicant (see 10.4).
of the cage or in back to front positions. Other food container
11.3 The reference animals shall be housed individually in
positions, such as side by side, are also permissible if they do
cages of the same or similar type and size as those used for the
not bias the test results.
actual test group.
13.5 To reduce feeding position biases, take the following
11.4 The reference animals may be placed on the same or a
precautions:
separate cage rack from the test animals and be housed in the
13.5.1 Once the animals have been placed in the cages, do
same animal room or they may be maintained in another
not change the position of the cages in the rack or the position
animal room under the same environmental conditions (that is,
of the rack in the room during the test period.
temperature, humidity, and lighting) as the test animals.
13.5.2 During the acclimation period, offer a commercial
11.5 The reference animal group shall be maintained con-
rodent diet in equal amounts, in excess of daily requirements,
currently with the test animal group. One group of reference
in both food containers. Food consumption should be measured
rodents may be used for one or more test groups in laboratories
during this period to establish baseline consumption for each
equipped to conduct several tests at one time, provided all tests
animal. A minimum of two days is recommended.
are with the same species and the rodents are obtained from the
13.5.3 On the day of the test, remove the food containers
same source at the same time (as one lot).
and replace them with clean containers, one contai
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