Appendixes

Appendix A

Testing Guidelines

Testing Guidelines

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, andto facilitate the work of those who design and carryout tests. Many organizations have developed testingguidelines. Three such compilations have been selectedfor discussion.

FDA Guidelines InvolvingWhole Animal Testing

To the extent possible, the Food and Drug Adminis-tration (FDA) makes its animal testing guidelines con-sistent throughout the agency and consistent with thoseof other agencies and organizations. However, specialuses of products require special testing, and guidanceis available from agency staff to help manufacturersmeet those requirements. In this table, tests that gen-erally can be considered common or standard toxico-logical tests usually used throughout the agency aregrouped together. Those that are more specific forevaluation of the safety of certain products are identi-fied with the FDA Center responsible for regulatingthat product.

I. Agency-wideA. General Toxicity

1. Acute oral—rodent, nonrodent2. Acute dermal—rodent, nonrodent3. Acute inhalation—rodent4. Subchronic oral—rodent, nonrodent5. Chronic oral—rodent, nonrodent6. Carcinogenicity —rodent7. Combined chronic/carcinogenicity —rodent

B. Specific Effects1. Dermal sensitization—guinea pig2. Dermal irritation—rabbit3. Eye irritation—rabbit4. Teratogenicity—rodent, rabbit5. Reproduction—rodent6. Absorption, distribution, metabolism,

elimination—rodent, nonrodent7. Neural-behavioral—rodent, rabbit

H. Center-orientedA. Human Drugs

1. Subchronic inhalation—rodent, nonrodent2. Subchronic dermal—rodent, nonrodent

3. Vaginal and rectal administration—rodent,nonrodent

4. Immunotoxicity—rodentB. Food Additives/Color Additives

I. Immunotoxicity—rodent2. Protein quality—rodent3. Vitamin D assay—rodent

C. Biologics1. All biologics administered by injection

a. Safety—guinea pigs, miceb. pyrogenicity–rabbits

2. Vaccinesa. Safety—mice, suckling mice,

chimpanzees, monkeys, guinea pigs,rabbits

b. Potency–guinea pigs, mice, monkeysc. Hypersensitivity-guinea pigsd. Toxicity—mice

3. Antitoxinsa. Potency—guinea pigs, mice

4. Toxinsa. Potency—mice

5. Toxoidsa. Potency—mice

6. Immune globulinsa. Potency—guinea pigs

7. Tuberculina. Safety—guinea pigsb. Potency—mice

D. Devices1. Corneal metabolism—rabbit2. Biomaterial implant—rabbit, primate, cat3. U.S.P. intracutaneous—rabbit

E. Cosmetics1. Primary skin irritation and corrosivity -

rabbit2. Phototoxicity—nude mouse, rabbit, guinea

pigF. New Veterinary Drugs

1.2.3.4,5.6.7.8.

Safety, efficacy—target speciesDrug tolerance—target speciesReproduction studies—target speciesTissue irritation—target speciesCombination drug—target speciesDrug disposition—target speciesRoute of administration—target speciesIntramammary infusion-dairy cows, goats

383

384 ● Alternatives to Animal Use in Research, Testing, and Education

OECD Guidelines InvolvingWhole Animal Testing

The Organization for Economic Cooperation and De-velopment (OECD) guidelines have wide acceptance inthe United States and abroad because of the MutualAcceptance of Data Decision (l). Under the terms ofthis decision, member countries of OECD must acceptdata generated in other countries if done so accordingto these guidelines. Animal tests contained in the guide-lines are listed below.

1. Effects on Biotic Systems202 Daphnia, acute immobilization test and re-

production test203 Fish, acute toxicity test204 Fish, prolonged toxicity test: 14 day study205 Avian dietary toxicity test206 Avian reproduction test

2. Degradation and Accumulation305A Bioaccumulation:

Test305B Bioaccumulation:305C Bioaccumulation:

Bioconcentration305D Bioaccumulation:305E Bioaccumulation:

3. Health Effects

Sequential Static Fish

Semi-static Fish TestTest for the Degree ofin FishStatic Fish TestFlow-through Fish Test

Short-Term Toxicology401 Acute oral toxicity402 Acute dermal toxicity403 Acute inhalation toxicity404 Acute dermal irritation/corrosion405 Acute eye irritation/corrosion406 Skin sensitization407 Repeated dose oral toxicity—rodent: 14/28

day408 Subchronic oral toxicity–rodent: 90 day409 Subchronic oral toxicity—nonrodent: 90 day410 Repeated dose dermal toxicity: 14/28 day411 Subchronic dermal toxicity: 90 day412 Repeated dose inhalation toxicity: 14/28 day413 Subchronic inhalation toxicity: 90 day414 Teratogenicity415 One-generation reproduction toxicity study416 Two-generation reproduction toxicity study417 Toxicokinetics418 Acute delayed neurotoxicity of

organophosphorous substances419 Subchronic delayed neurotoxicity of

organophosphorous substances: 90 dayLong-Term Toxicology451 Carcinogenicity studies

452 Chronic toxicity studies453 Combined chronic toxicity/carcinogenicity

studiesGenetic Toxicology474 Genetic toxicity: micronucleus test475 In vivo mammalian bone marrow

cytogenetic test—chromosomal analysis478 Rodent dominant lethal test

Pesticide Assessment GuidelinesInvolving Whole-Animal Testing

The Office of Pesticide Programs of the Environmen-tal Protection Agency (EPA) has developed guidelinesfor testing required under the Federal Insecticide, Fun-gicide, and Rodenticide Act. These Pesticide AssessmentGuidelines contain standards for conducting acceptabletests, 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 EPA’s Office ofToxic Substances (OTS) for testing required under theToxic Substances Control Act (3).

Subdivision E: Hazard Evaluation: Wildlife andAquatic Organisms

Series 70: General Information and RequirementsSeries 71: Avian and Mammalian Testing

71-1 Avian single-dose oral LD5O test71-2 Avian dietary LC5O test71-3 Wild mammal toxicity test71-4 Avian reproduction test71-5 Simulated and actual field tests for mammals

and birdsSeries 72: Aquatic Organism Testing

72-1 Acute toxicity test for freshwater fish72-2 Acute toxicity test for freshwater aquatic

invertebrates72-3 Acute toxicity test for estuarine and marine

organisms72-4 Fish early life-stage and aquatic invertebrate

life-cycle studies72-5 Life-cycle tests of fish72-6 Aquatic organism accumulation tests72-7 Simulated or actual field testing for aquatic

organisms

Subdivision F: Hazard Evaluation: Humansand Domestic Animals

Series 80: Overview, Definition, and Genera]Requirements

Series 81: Acute Toxicity and Irritation Studies81-1 Acute oral toxicity study

App. A—Testing Guidelines ● 385

81-2 Acute dermal toxicity study81-3 Acute inhalation toxicity study81-4 Primary eye irritation study81-5 Primary dermal irritation study81-6 Dermal sensitization study81-7 Acute delayed neurotoxicity of

Series82-1

82-282-382-482-5

Series83-183-283-383-483-5

Series84-1

84-2

Series85-185-2

organophosphorous substances82: Subchronic TestingSubchronic oral toxicity (rodent andnonrodent): 90 day studyRepeated dose dermal toxicity: 21 day studySubchronic dermal toxicity: 90 day studySubchronic inhalation toxicity: 90 day studySubchronic neurotoxicity: 90 day study83: Chronic and Long-Term StudiesChronic toxicity studiesOncogenicity studiesTeratogenicity studyReproductive and fertility effectsCombined chronic toxicity/oncogenicitystudies84: MutagenicityPurpose and general recommendations formutagenicity testingMutagenicity tests (described in very generalterms, with reference to the OTS guidelines)85: Special StudiesMetabolism studyDomestic animal safety testing

Subdivision G: Product Performance

Series 95: Efficacy of Invertebrate Control Agents95-1 Genera] considerations95-8 Livestock, poultry, fur and wool-bearing ani-

mal treatments95-9 Treatments to control pests of humans and

petsSeries 96: Efficacy of Vertebrate Control Agents

96-196-296-396-4

96-596-696-796-896-9

General considerationsFish control agentsAquatic amphibian control agentsTerrestrial amphibian and reptilian controlagentsAvian toxicantsAvian repellentsAvian frightening agentsMole toxicantsBat toxicants and repellents

96-10 Commensal rodenticides96-11 Rodenticides in orchards96-12 Rodenticides on farm and rangelands96-13 Rodent fumigants96-14 Rodent repellents on tree seeds96-15 Rodent repellents on cables96-16 Rodent reproductive inhibitors96-17 Mammalian predacides96-18 Domestic dog and cat repellents96-19 Browsing animal repellents96-30 Methods and protocols

Subdivision M: Biorational Pesticides

(This subdivision duplicates many of the provisionsof other subdivisions, and is therefore not describedin detail. )Series 150: Overview, Definitions, and General

ProvisionsSeries 152: Toxicology Guidelines

Subseries 152A: Toxicology GuidelinesSubseries 152B: Toxicology Guidelines for

Microbial Pest Control AgentsSeries 154: Nontarget Organism Hazard Guidelines

Subseries 154A: Nontarget Organism HazardGuidelines for BiochemicalAgents

Subseries 154B: Nontarget Organism HazardGuidelines for Microbial Agents

Series 157: Experimental Use Permit Guidelines

Subdivision N: Environmental Fate

Series 165: Accumulation Studies165-4 Laboratory Studies of Pesticide

Accumulation in Fish165-5 Field Accumulation Studies of Aquatic

Nontarget Organisms

Appendix 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).