Appendixes

Appendix A
Testing Guidelines
Testi ng Gui del i nes
Testing guidelines are developed for a variety of rea-
sons: to allow results of various test substances or spe-
cies to be easily compared, to encourage the use of cer-
tain protocols so that testing need not be repeated, and
to facilitate the work of those who design and carry
out tests. Many organizations have developed testing
guidelines. Three such compilations have been selected
for discussion.
FDA Gui del i nes I nvol vi ng
Whol e Ani mal Testi ng
To the extent possible, the Food and Drug Adminis-
tration (FDA) makes its animal testing guidelines con-
sistent throughout the agency and consistent with those
of other agencies and organizations. However, special
uses of products require special testing, and guidance
is available from agency staff to help manufacturers
meet those requirements. In this table, tests that gen-
erally can be considered common or standard toxico-
logical tests usually used throughout the agency are
grouped together. Those that are more specific for
evaluation of the safety of certain products are identi-
fied with the FDA Center responsible for regulating
that product.
I. Agency-wide
A. General Toxicity
1. Acute oralrodent, nonrodent
2. Acute dermalrodent, nonrodent
3. Acute inhalationrodent
4. Subchronic oralrodent, nonrodent
5. Chronic oralrodent, nonrodent
6. Carcinogenicity rodent
7. Combined chronic/carcinogenicity rodent
B. Specific Effects
1. Dermal sensitizationguinea pig
2. Dermal irritationrabbit
3. Eye irritationrabbit
4. Teratogenicityrodent, rabbit
5. Reproductionrodent
6. Absorption, distribution, metabolism,
eliminationrodent, nonrodent
7. Neural-behavioralrodent, rabbit
H. Center-oriented
A. Human Drugs
1. Subchronic inhalationrodent, nonrodent
2. Subchronic dermalrodent, nonrodent
3. Vaginal and rectal administrationrodent,
nonrodent
4. Immunotoxicityrodent
B. Food Additives/Color Additives
I. Immunotoxicityrodent
2. Protein qualityrodent
3. Vitamin D assayrodent
C. Biologics
1. All biologics administered by injection
a. Safetyguinea pigs, mice
b. pyrogenicityrabbits
2. Vaccines
a. Safetymice, suckling mice,
chimpanzees, monkeys, guinea pigs,
rabbits
b. Potencyguinea pigs, mice, monkeys
c. Hypersensitivity-guinea pigs
d. Toxicitymice
3. Antitoxins
a. Potencyguinea pigs, mice
4. Toxins
a. Potencymice
5. Toxoids
a. Potencymice
6. Immune globulins
a. Potencyguinea pigs
7. Tuberculin
a. Safetyguinea pigs
b. Potencymice
D. Devices
1. Corneal metabolismrabbit
2. Biomaterial implantrabbit, primate, cat
3. U.S.P. intracutaneousrabbit
E. Cosmetics
1. Primary skin irritation and corrosivity -
rabbit
2. Phototoxicitynude mouse, rabbit, guinea
pig
F. New Veterinary Drugs
1.
2.
3.
4,
5.
6.
7.
8.
Safety, efficacytarget species
Drug tolerancetarget species
Reproduction studiestarget species
Tissue irritationtarget species
Combination drugtarget species
Drug dispositiontarget species
Route of administrationtarget species
Intramammary infusion-dairy cows, goats
383
384 G Alternatives to Animal Use in Research, Testing, and Education
OECD Gui del i nes I nvol vi ng
Whol e Ani mal Testi ng
The Organization for Economic Cooperation and De-
velopment (OECD) guidelines have wide acceptance in
the United States and abroad because of the Mutual
Acceptance of Data Decision (l). Under the terms of
this decision, member countries of OECD must accept
data generated in other countries if done so according
to these guidelines. Animal tests contained in the guide-
lines are listed below.
1. Effects on Biotic Systems
202 Daphnia, acute immobilization test and re-
production test
203 Fish, acute toxicity test
204 Fish, prolonged toxicity test: 14 day study
205 Avian dietary toxicity test
206 Avian reproduction test
2. Degradation and Accumulation
305A Bioaccumulation:
Test
305B Bioaccumulation:
305C Bioaccumulation:
Bioconcentration
305D Bioaccumulation:
305E Bioaccumulation:
3. Health Effects
Sequential Static Fish
Semi-static Fish Test
Test for the Degree of
in Fish
Static Fish Test
Flow-through Fish Test
Short-Term Toxicology
401 Acute oral toxicity
402 Acute dermal toxicity
403 Acute inhalation toxicity
404 Acute dermal irritation/corrosion
405 Acute eye irritation/corrosion
406 Skin sensitization
407 Repeated dose oral toxicityrodent: 14/28
day
408 Subchronic oral toxicityrodent: 90 day
409 Subchronic oral toxicitynonrodent: 90 day
410 Repeated dose dermal toxicity: 14/28 day
411 Subchronic dermal toxicity: 90 day
412 Repeated dose inhalation toxicity: 14/28 day
413 Subchronic inhalation toxicity: 90 day
414 Teratogenicity
415 One-generation reproduction toxicity study
416 Two-generation reproduction toxicity study
417 Toxicokinetics
418 Acute delayed neurotoxicity of
organophosphorous substances
419 Subchronic delayed neurotoxicity of
organophosphorous substances: 90 day
Long-Term Toxicology
451 Carcinogenicity studies
452 Chronic toxicity studies
453 Combined chronic toxicity/carcinogenicity
studies
Genetic Toxicology
474 Genetic toxicity: micronucleus test
475 In vivo mammalian bone marrow
cytogenetic testchromosomal analysis
478 Rodent dominant lethal test
Pesti ci de Assessment Gui del i nes
I nvol vi ng Whol e-Ani mal Testi ng
The Office of Pesticide Programs of the Environmen-
tal Protection Agency (EPA) has developed guidelines
for testing required under the Federal Insecticide, Fun-
gicide, and Rodenticide Act. These Pesticide Assessment
Guidelines contain standards for conducting acceptable
tests, guidelines for the evaluation and reporting of data,
guidelines as to when additional testing might be re-
quired, and examples of acceptable protocols (2). Simi-
lar guidelines have been developed by EPAs Office of
Toxic Substances (OTS) for testing required under the
Toxic Substances Control Act (3).
Subdivision E: Hazard Evaluation: Wildlife and
Aqu at ic Or gan is ms
Series 70: General Information and Requirements
Series 71: Avian and Mammalian Testing
71-1 Avian single-dose oral LD
5O
test
71-2 Avian dietary LC
5O
test
71-3 Wild mammal toxicity test
71-4 Avian reproduction test
71-5 Simulated and actual field tests for mammals
and birds
Series 72: Aquatic Organism Testing
72-1 Acute toxicity test for freshwater fish
72-2 Acute toxicity test for freshwater aquatic
invertebrates
72-3 Acute toxicity test for estuarine and marine
organisms
72-4 Fish early life-stage and aquatic invertebrate
life-cycle studies
72-5 Life-cycle tests of fish
72-6 Aquatic organism accumulation tests
72-7 Simulated or actual field testing for aquatic
organisms
Su bdivis ion F: Hazar d Evalu at ion : Hu man s
an d Domes t ic An imals
Series 80: Overview, Definition, and Genera]
Requirements
Series 81: Acute Toxicity and Irritation Studies
81-1 Acute oral toxicity study
App. ATesting Guidelines G 385
81-2 Acute dermal toxicity study
81-3 Acute inhalation toxicity study
81-4 Primary eye irritation study
81-5 Primary dermal irritation study
81-6 Dermal sensitization study
81-7 Acute delayed neurotoxicity of
Series
82-1
82-2
82-3
82-4
82-5
Series
83-1
83-2
83-3
83-4
83-5
Series
84-1
84-2
Series
85-1
85-2
organophosphorous substances
82: Subchronic Testing
Subchronic oral toxicity (rodent and
nonrodent): 90 day study
Repeated dose dermal toxicity: 21 day study
Subchronic dermal toxicity: 90 day study
Subchronic inhalation toxicity: 90 day study
Subchronic neurotoxicity: 90 day study
83: Chronic and Long-Term Studies
Chronic toxicity studies
Oncogenicity studies
Teratogenicity study
Reproductive and fertility effects
Combined chronic toxicity/oncogenicity
studies
84: Mutagenicity
Purpose and general recommendations for
mutagenicity testing
Mutagenicity tests (described in very general
terms, with reference to the OTS guidelines)
85: Special Studies
Metabolism study
Domestic animal safety testing
Su bdivis ion G: Pr odu ct Per for man ce
Series 95: Efficacy of Invertebrate Control Agents
95-1 Genera] considerations
95-8 Livestock, poultry, fur and wool-bearing ani-
mal treatments
95-9 Treatments to control pests of humans and
pets
Series 96: Efficacy of Vertebrate Control Agents
96-1
96-2
96-3
96-4
96-5
96-6
96-7
96-8
96-9
General considerations
Fish control agents
Aquatic amphibian control agents
Terrestrial amphibian and reptilian control
agents
Avian toxicants
Avian repellents
Avian frightening agents
Mole toxicants
Bat toxicants and repellents
96-10 Commensal rodenticides
96-11 Rodenticides in orchards
96-12 Rodenticides on farm and rangelands
96-13 Rodent fumigants
96-14 Rodent repellents on tree seeds
96-15 Rodent repellents on cables
96-16 Rodent reproductive inhibitors
96-17 Mammalian predacides
96-18 Domestic dog and cat repellents
96-19 Browsing animal repellents
96-30 Methods and protocols
Su bdivis ion M: Bior at ion al Pes t icides
(This subdivision duplicates many of the provisions
of other subdivisions, and is therefore not described
in detail. )
Series 150: Overview, Definitions, and General
Provisions
Series 152: Toxicology Guidelines
Subseries 152A: Toxicology Guidelines
Subseries 152B: Toxicology Guidelines for
Microbial Pest Control Agents
Series 154: Nontarget Organism Hazard Guidelines
Subseries 154A: Nontarget Organism Hazard
Guidelines for Biochemical
Agents
Subseries 154B: Nontarget Organism Hazard
Guidelines for Microbial Agents
Series 157: Experimental Use Permit Guidelines
Su bdivis ion N: En vir on men t al Fat e
Series 165: Accumulation Studies
165-4 Laboratory Studies of Pesticide
Accumulation in Fish
165-5 Field Accumulation Studies of Aquatic
Nontarget Organisms
Appendi x A References
1. Organization for Economic Cooperation and Development,
Guidelines for Testing of Chemicals, and addenda (Paris:
1981).
2. US. Environmental Protection Agency, Pesticide Assess-
ment Guidelines (Springfield, VA: National Technical In-
formation Service, 1984).
3. U.S. Environmental Protection Agency, Office of Toxic Sub-
stances Health and Environmental Effects Test Guidelines
(Washington, DC: update October 1984).