GUIDE TO THE CARE AND USE OF EXPERIMENTAL ANIMALS

Edited by: Ernest D. Olfert, DVM; Brenda M. Cross, DVM; and A. Ann McWilliam

CCAC, Guide Vol. 1 (2nd Ed.) 1993

Volume 1, 1993

Editors:
Dr E.D. Olfert
Director
Animal Resources Centre
University of Saskatchewan
Saskatoon, Saskatchewan S7N 0W0

Dr B.M. Cross
Asssistant Director
Animal Resources Centre
University of Saskatchewan
Saskatoon, Saskatchewan S7N 0W0

Mrs A.A. McWilliam

Information Officer
Canadian Council on Animal Care
1000-151 Slater Street
Ottawa, Ontario K1P 5H3

Canadian Council on Animal Care, 1993

ISBN: 0-919087-18-3

Citing certain devices or manufacturers is not to be perceived as the endorsement of the

Canadian Council on Animal Care (CCAC) of one particular product over another.

In keeping with the CCAC policy of revising statements and

guidelines as needed, users of this Guide are encouraged to forward
any comments to the Secretariat.

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 Table of Contents
DEDICATION
PREFACE
FOREWORD
ACKNOWLEDGEMENTS
CONTRIBUTORS
LIST OF ABBREVIATIONS

I.
RESPONSIBILITY FOR THE CARE AND USE OF
EXPERIMENTAL ANIMALS
A. NATIONAL LEVEL
1. Evolution of the Canadian Council on Animal Care

2. The Contemporary Council

3. CCAC's Assessment Program

4. CCAC Position Statements

5. Legislation Governing Experimental Animals

6. Pre-University Use of Animals

B. LOCAL LEVEL
1. The Institutional Animal Care Committee
2. Terms of Reference for Animal Care Committees

3. The Veterinarian

4. Animal Care Personnel

C. REFERENCES

II.
LABORATORY ANIMAL FACILITIES
A. INTRODUCTION
B. LOCATION
C. MECHANICAL SERVICES
D. DESIGN
E. MAJOR FUNCTIONAL DIVISIONS
1. Animal Reception Area
2. Conditioning Rooms
3. Holding Rooms
4. Quarantine/Isolation Rooms
5. Experimental and Treatment Facilities

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 6. Support Facilities
7. Personnel, Office and Reception Areas
8. Facilities for Personnel

F. SECURITY
G. CONSTRUCTION GUIDELINES FOR ANIMAL ROOMS
1. Floors and Drains
2. Walls and Ceilings
3. Doors
4. Windows
5. Corridors
6. Services

H. CAGING
1. Shoebox Cages
2. Larger Solid Bottom Caging
3. Suspended Cages
4. Other Cages

I. REFERENCES

III.
THE ENVIRONMENT
A. CLIMATE CONTROL
1. Temperature
2. Humidity
3. Ventilation
4. Lighting

B. OTHER ENVIRONMENTAL FACTORS

1. Noise

2. Chemicals

3. Bedding

4. Population Density and Space Limitations

C. MICROBIOLOGICAL CONTROL
1. Conventional Facilities

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 2. Barrier Facilities
3. Biohazard Containment

D. CHEMICAL AND RADIOISOTOPE UNITS

E. REFERENCES

IV.
FARM ANIMAL FACILITIES AND ENVIRONMENT
A. FACILITIES
B. SPECIFIC ENVIRONMENTAL CONSIDERATIONS
1. Cattle
2. Sheep
3. Swine
4. Horses
5. Poultry

C. PEST CONTROL
D. REFERENCES

V.
LABORATORY ANIMAL CARE
A. INTRODUCTION
B. GENERAL PRACTICES
1. Reception

2. Conditioning/Quarantine
3. Holding (Maintenance)
4. Identification and Records

C. CARE OF THE ANIMAL

1. Food
2. Water
3. Exercise

D. CARE OF THE FACILITY

1. Cleaning and Sanitation
2. Waste Disposal
3. Vermin Control
4. Holiday and Emergency Care

E. REFERENCES

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VI.
SOCIAL AND BEHAVIORAL REQUIREMENTS OF
EXPERIMENTAL ANIMALS
A. INTRODUCTION
1. What is Animal Well-being or Welfare?
2. Environmental Enrichment
3. Group Formation
4. Position Statement

B. ANIMALS USED IN AGRICULTURAL RESEARCH

1. Introduction
2. Animal Stress
3. Housing and Husbandry
4. General Principles

C. ANIMALS (LARGE) HELD IN METABOLISM CAGES

1. Conditioning
2. Size of Metabolism Crates
3. Contact with Other Animals
4. Pre-, During and Post-Experiment Checks
5. Observing Changes in Behavior
6. Duration of Confinement
7. Exceptional Circumstances

D. CATS
1. Introduction
2. Behavioral Enrichment
3. Social Peers
4. Enrichment Devices (Artificial Appliances)
5. Food Gathering Activities
6. Control of the Environment
7. Housing
8. Maternal Behavior
9. Random-Source vs. Purposebred Animals

E. DOGS
1. Introduction

2. Breed Differences
3. Criteria for Assessing Well-being

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 4. Housing

5. Socialization to People
6. Enrichment Devices (Artificial Appliances)
7. Exercise

F. NON-HUMAN PRIMATES
1. Introduction
2. Interpretation of the Behavioral and Morphological Postures
3. Distinctive Characteristics
4. Assessing Social and Behavioral Well-being
5. Ways of Promoting Social and Behavioral Well-being
6. Disposition

7. Summary

G. RODENTS AND RABBITS

1. Introduction

2. Behavioral Enrichment and Social Peers

3. Enrichment Devices (Artificial Appliances)

4. Caging and Bedding

5. Food Gathering

6. Control of the Environment

H. WILDLIFE HELD IN THE LABORATORY

I. REFERENCES

VII.
SPECIAL PRACTICES
A. ANIMAL ACQUISITION
1. Procurement

2. Transportation

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 3. Breeding

4. Breeding Transgenic Animals

5. Animal Models with Special Needs

6. Identification of the Sexes

B. RESTRAINT AND MANIPULATIONS

1. Physical Restraint

2. Implantation, Cannulation and Sampling

3. Bleeding

4. Motivation Procedures

C. REFERENCES

VIII.
OCCUPATIONAL HEALTH AND SAFETY
A. REGULATORY REQUIREMENTS
B. BILOGICAL HAZARDS
C. ZOONOSES
D. PROCEDURES FOR WORKING WIHT NON-HUMAN PRIMATES
E. ALLERGIES
F. PHYSICAL INJURIES AND CHEMICAL HAZARDS
G. RADIATION AND ULTRAVIOLET LIGHT
H. REFERENCES

IX.
STANDARDS FOR EXPERIMENTAL ANIMAL SURGERY
A. INTRODUCTION
B. FACILITIES FOR SURVIVAL SURGERY
C. PRE-OPERATIVE PLANNING AND ANIMAL PREPARATION
D. SURGICAL PROCEDURES AND INTRA-OPERATIVE NURSING
CARE
E. POST-OPERATIVE RECOVERY AND SUPPORT
F. REFERENCES

X.

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 CONTROL OF ANIMAL PAIN IN RESEARCH, TEACHING
AND TESTING
A. INTRODUCTION
B. WHAT IS ANIMAL PAIN?
C. GUIDELINES
D. THE ROLE OF THE VETERINARIAN IN REDUCING PAIN
E. SIGNS OF PAIN AND DISTRESS
F. ANALGESIC AGENTS
1. Opioid Agonists
2. Opioid Agonist/Antagonists
3. Opioid Antagonists
4. Non-steroidal Anti-inflammatory Drugs (NSAIDS)
5. Analgesia provided by Local Anesthetics
6. Neuroleptanalgesics

G. AREAS FOR FUTURE STUDY

H. REFERENCES

XI.
ANESTHESIA
A. MANAGEMENT OF ANESTHESIA
1. General
2. Handling the Patient
3. Fasting
4. Anticholinergics

B. TRANQUILLIZERS AND SEDATIVES

C. GENERAL ANESTHETICS
1. Dissociative Anesthetics
2. Barbiturates
3. Chloralose
4. Urethane (Urethan, Ethyl Carbamate)
5. SaffanTM
6. Tribomoethanol (Avertin)
7. Non-specific Injectable Anesthetic Antagonists
8. Inhalant Anesthetics

D. MUSCLE RELAXANTS
1. Glyceryl Guiacolate

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 2. Neuromuscular Blocking Agents

E. LOCAL AND REGIONAL ANESTHETICS

F. ANIMAL HYPNOSIS (TONIC IMMOBILITY)
G. SPECIES CONSIDERATIONS
H. REFERENCES

XII. EUTHANASIA
A. INTRODUCTION
B. CRITERIA FOR A HUMANE DEATH
C. PAIN AND STRESS
D. MECHANISMS FOR CAUSING DEATH
E. METHODS USED FOR EUTHANASIA
1. Physical
2. Non-inhalant Pharmacologic Agents
3. Inhalant Anesthetics
4. Non-anesthetic Gases

F. SPECIFIC SPECIES
1. Amphibians, Fishes and Reptiles
2. Domestic Animals Killed for Food
3. Fur-bearing Animals

G. TISSUE EFFECTS OF EUTHANASIA METHODS

1. Direct Effects
2. Indirect Effects

H. EFFECT ON OBSERVERS
I. EUTHANASIA STATEMENTS--OTHER AGENCIES
J. REFERENCES

XIII.
THE USE OF ANIMALS IN PSYCHOLOGY

A. THE SCIENTIST
B. RESEARCH
C. INSTRUCTION
D. REFERENCES

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XIV.
GUIDELINES FOR THE USE OF ANIMALS IN
NEUROSCIENCE RESEARCH

A. INTRODUCTION
B. FACTORS THAT RELATE TO THE DESIGN OF EXPERIMENTS
C. FACTORS THAT RELATE TO THE CONDUCT OF EXPERIMENTS

APPENDICES

I. Housing and Environment

II. Breeding and Reproduction Data
III. Physiological and Nutritional Parameters
IV. Hematology
V. Clinical Biochemistry Reference Values
VI. Serum Electrolyte Reference Values
VII. Zoonoses--Experimental Animals to Man
A. Bacterial Diseases
B. Rickettsial Diseases

C. Arbovirus Diseases

D. Other Virus Diseases

E. Fungal and Protozoan Diseases

VIII. Common Bleeding Sites

IX. Tranquillizer, Sedative and Anticholinergic Drug Dosages
X. Analgesic Drug Dosages
XI. Injectable Anesthetic Agents--Dosage
XII. Anesthetic and Sedative Drug Dosage--Amphibians and Reptiles
XIII. Anesthetic and Sedative Drug Dosage--Fishes
XIV. Methods for Euthanasia by Species
XV. CCAC Position Statements
A. Ethics of Animal Investigation
B. Categories of Invasiveness in Animal Experiments
C. CCAC Guidelines on Acceptable Immunological Procedures

XVI. Journals Held by CCAC

XVII. Glossary

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 I. RESPONSIBILITY FOR THE CARE AND USE OF
EXPERIMENTAL ANIMALS

All experimental care and use of animals in this country is subject to the
requirements of the Canadian Council on Animal Care (CCAC), a national, peer
review organization founded in Ottawa in 1968. Its mandate is straightforward
and concise:

"to work for the improvement of animal care and use on a Canada-wide basis".

A. NATIONAL LEVEL

1. Evolution of the Canadian Council on Animal Care

The 1950s and 1960s were a period of phenomenal growth in Canadian research and
graduate teaching, particularly in biomedical sciences. Expression of public concern over
the use of animals in research also increased during this period, as did the awareness of
the scientific community that this constituted a sensitive area, and raised ethical
questions, not the least of which was responsibility for animal care and use.

In 1963, the Medical Research Council (MRC) decided that the matter warranted further
study, and the following year it requested that the National Research Council (NRC)
establish a committee to investigate the care and use of experimental animals in Canada.
The report of the Special Committee on the Care of Experimental Animals (1966)
recommended the creation of a voluntary control program exercised by scientists in each
institution, subject to peer judgement and committed to implement the guiding principles
of an independent advisory body.

A feasibility study of these proposals was undertaken (Rowsell, 1967) and, as a result, all
universities and government departments where animals were used agreed to support the
formation of a Canadian council on animal care (Anon., 1967). The CCAC was
established in 1968 as a standing committee of the Association of Universities and
Colleges of Canada (AUCC). Comprising 12 member organizations, including the
Canadian Federation of Humane Societies (CFHS), its terms of reference included the
making of recommendations for improvements in:

a) the procurement and production of experimental animals;

b) the facilities and care of experimental animals;
c) the control over experiments involving animals.

At its inaugural meeting, held January 30, 1968, the CCAC adopted as its objective: "to
develop guiding principles for the care of experimental animals in Canada, and to work
for their effective application." A Resources Panel and an Animal Care Review Panel
were created to assist the secretariat in achieving its objective. The functions of the
original resources panel have, in recent years, been assumed by specific committees

11
responsible for the development and implementation of national policies on laboratory
animal resources.

Prior to CCAC's establishment, the only guidelines which had been defined for the care
and use of experimental animals in Canada had been the Canadian Federation of
Biological Societies' (CFBS) one-page Guiding Principles for the Care of Experimental
Animals (1961). Within its first year, the Council published comprehensive guidelines
entitled Care of Experimental Animals: A Guide for Canada. It demanded use of non-
sentient methods wherever possible, and noted that "when an animal must be used there
is an obligation to:

a) provide humane care and treatment;

b) minimize pain and discomfort;
c) avoid unnecessary use."

2. The Contemporary Council

Co-funded by this country's two major granting agencies, the MRC and the Natural
Sciences and Engineering Research Council (NSERC), the CCAC comprises 20 member
organizations, whose representatives include scientists, educators, and representatives of
industry and the animal welfare movement. They include:

1. Agriculture Canada (AC)

2. Association of Canadian Faculties of Dentistry (ACFD)
3. Association of Canadian Medical Colleges (ACMC)
4. Association of Universities and Colleges of Canada (AUCC)
5. Canadian Association for Laboratory Animal Medicine (CALAM)
6. Canadian Association for Laboratory Animal Science (CALAS)
7. Canadian Federation of Humane Societies (CFHS)
8. Committee of Chairpersons of Departments of Psychology (CCDP)
9. Canadian Society of Zoologists (CSZ)
10. Confederation of Canadian Faculties of Agriculture and Veterinary Medicine
(CCFAVM)
11. Department of National Defence (DND)
12. Environment Canada (EC)
13. Fisheries and Oceans Canada (FOC)
14. Health and Welfare Canada (HWC)
15. Canadian Heart and Stroke Foundation of Canada (HSFC)
16. Medical Research Council (MRC)
17. National Cancer Institute (NCI)
18. National Research Council (NRC)
19. Natural Sciences and Engineering Research Council (NSERC)
20. Pharmaceutical Manufacturers' Association of Canada (PMAC)

CCAC assessment panels evaluate animal care and use in Canadian universities and
community colleges, government laboratories, and commercial laboratories. The effects

12
of the CCAC program have included improvement in both housing and management
practices. As a result, although the research community is growing, the numbers of
experimental animals has, to date, steadily declined; however, they may increase in future
because of increased use of transgenic animals and genetic manipulation. This has
already occurred in the U.K. (Anon., 1992d).

The concerns of the Council in such areas as alternatives, animal biotechnology, facilities
standards, immunological procedures, invertebrates and wild vertebrates are reflected in
its committees which comprise experts from a wide variety of biological disciplines and
the animal welfare community.

3. CCAC's Assessment Program

The CCAC carries out its national responsibility for animal care through education in the
form of workshops, publications, presentations, etc., and its assessment program, which
focuses on animal care and use, and the evaluation of the effectiveness of local Animal
Care Committees (ACC). These institutional committees are responsible for assuring
ethical animal use and compliance with CCAC guidelines at the local level, and must
evaluate the ethical aspects of proposed research before the study may commence.
(CCAC Recommended Terms of Reference and Guidelines for Animal Care Committees
are found later in this chapter.) Assessments are based on Volumes 1 and 2 of this Guide
and CCAC position papers. In-depth assessments are normally scheduled approximately
every three years. In addition, a number of Special or Unannounced Visits are conducted
if a panel and/or the Council feels that the conditions at an institution so warrant, or upon
request by the institution.

a) The Panel

The scientific members of an assessment panel are chosen by CCAC from an inventory
of individuals with experience and special knowledge of various aspects of animal
experimentation and care; they are selected, as much as possible, with reference to the
predominant thrust of the research at the institution to be assessed. As well, each panel
includes a CFHS or other animal welfare appointee, usually drawn from the geographic
area of the institution to be visited. All panellists serve voluntarily and without
remuneration except expenses. The average complement of a panel is three scientists and
one animal welfare representative, with the Director or Associate Director of Assessments
serving as an ex-officio member. It is sometimes necessary in the larger institutions to
increase the size of the panel and divide it into sub-groups. In some of the larger
institutions, assessments may be carried out on a faculty or departmental basis.

b) Preparation for the Site Visit

Prior to each assessment visit, the Council requests and receives from the institution
current information pertaining to:

i) administrative organization;

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ii) animal care personnel;
iii) space allocation and location(s) for animal housing and use;
iv) animal use.

The institution also provides an animal use summary indicating the species and numbers
of animals being used, and an indication of the research or teaching project in which they
are involved. This permits the panel to give notice of specific projects members may wish
to examine in depth, including direct discussion with the principal investigator. Statistics
on animal use retained by CCAC are now updated annually through the use of the Animal
Research Protocol Management System (ARPMS) computer program developed and
provided to each institution by CCAC.

c) The Assessment

Most assessments begin with a meeting with the ACC and senior administrative
personnel of the institution. At this time, the institution's animal care program is
reviewed, and its response to the previous assessment panel's recommendations
discussed. Other topics might include changes to facilities, changes in personnel,
occupational health and safety programs, major changes in animal use, the "state-of-the-
art" of animal care, and the importance of environmental enrichment.

A considerable amount of time is spent with the institutional ACC reviewing terms of
reference, minutes from meetings and discussing the protocol approval process to ensure
the committee is functioning effectively.

All areas which house or hold animals are visited, as are all areas in which procedures on
animals are performed, e.g., surgical suites and laboratory testing areas. During these
visits, many individual investigators are interviewed, and specific procedures or
techniques may be observed. The panel is usually accompanied on the site visit by a
member(s) of the ACC, and pertinent departmental officials.

Following the site visit, a meeting with the ACC and senior officials is reconvened. This
is particularly important if the panel has major or serious concerns about any aspect of
the animal care and use; if so, the panel will request immediate appropriate action if a
particular situation so warrants. A general outline of the panel's observations and Major
or Serious recommendations, which will be forthcoming in an in-depth report, is usually
provided at this time.

Deficiencies which are of a Major nature, i.e., where an animal's health is at risk, must be
addressed immediately. Failure to take action would further jeopardize animal welfare
and could place an institution in a status of Non-compliance.

Institutions are given only short periods of time (e.g., three months) to respond to
recommendations of a Serious nature. Regular recommendations, which most commonly
concern housekeeping or facility maintenance, must be implemented at least by the time
of the next in-depth assessment.

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d) Relevant Reports

A subsequent in-depth report and recommendations, prepared by the panel, are aimed at
helping the institution to improve its animal care practices and/or facilities to a standard
in keeping with this Guide. These reports are circulated to the members of the CCAC
prior to being forwarded to the senior administrative official of the institution. The report
and its contents are considered confidential; however, the institutional official(s) may
distribute copies to whom they wish. If they wish to release the report to the public, the
CCAC should have prior notification. Occasionally, a panel member may have concerns
which do not reflect the view of the majority of the panel. In such cases, this member has
the right to submit a minority report.

In response to the panel's report, institutions are asked to submit to the CCAC within six
months a report describing the methods it proposes employing in order to implement the
report's recommendations. Should this implementation report be considered
unsatisfactory by the Council, the CCAC may instruct its secretariat to determine the
reasons for non-compliance and to take such further actions as deemed necessary. For
example, the CCAC notifies the MRC and NSERC of any institution found to be in non-
compliance with CCAC standards, and which has not responded satisfactorily within the
time given to correct the situation. A statement issued in March, 1985, notes:

"On receipt of a statement on non-compliance, and after reviewing the full evidence,
NSERC and MRC reserve the right, either separately or together, to bring their
concerns to the appropriate authorities in the research institution concerned and, if
they deem it necessary, to implement such financial or other sanctions as may be in
the power of either Council. Such sanctions may be taken whether or not the non-
compliance concerns research funded by either Council and might include the
freezing or withdrawal of research funds for any or all research programs funded by
either or both Research Councils in the institution."

e) Summary

CCAC's programs have been well received, and enjoy the support and co-operation of all
institutions involved, including private sector companies and government departments
that are not dependent on funds from granting agencies. It has also resulted in an
increased level of awareness and sensitivity to the ethics of animal experimentation
amongst scientists and investigators. The Council continues to encourage the use of
alternatives to animal use wherever possible, as exemplified by Russell and Burch's "3R"
tenet of Replacement, Reduction and Refinement (Russell and Burch, 1959; Smythe,
1978).

4. CCAC Position Statements

In addition to the Guide, (Volumes 1 and 2) the CCAC develops and publishes position
statements on a number of matters. Such position statements are subject to periodic

15
revision. These include Ethics of Animal Investigation, CCAC Guidelines on Acceptable
Immunological Procedures, and Categories of Invasiveness in Animal Experiments,
which may be found in the Appendices. The Social and Behavioural Requirements of
Experimental Animals (SBREA) document may be found elsewhere in this Guide. As
noted, these position statements, along with the Guide, form the basis on which CCAC
assessments are made.

5. Legislation Governing Experimental Animals

Animal welfare is governed in Canada under Section 446 of the Criminal Code.
Legislation pertaining specifically to experimental animals exists in two provincial acts:
Animals for Research Act (Ontario) and Universities Act (Alberta). There are numerous
other laws that impact on animals kept for research, teaching and testing.

a) Federal Legislation

The Criminal Code of Canada, Section 446, Cruelty to Animals, forbids "causing
unnecessary suffering." The century-old (1892) Code states that: "Everyone commits an
offence who wilfully causes or, being the owner, wilfully permits to be caused
unnecessary pain, suffering or injury to an animal or bird...."

The Law Reform Commission of Canada (LRCC), after 15 years' preparation (Anon.,
1988a), and considerable public input, proposed widespread amendments to the Criminal
Code in a report entitledReport 31 Recodifying Criminal Law in 1988 (LRCC, 1987;
Anon., 1988b). The proposed changes relating to animals centred on treating animals as
sentient beings, and not merely chattel put here for humans' use, a position the Canadian
Veterinary Medical Association (CVMA), among others, considered merited support
(Olfert, Finley, Laniel et al. 1989). Chapter 20, Title IV, Crimes Against the Natural
Order, Crimes Against Animals, focused on three areas: cruelty to animals, organizing
sporting events and animal neglect. It noted, for example, that "Everyone commits a
crime who unnecessarily causes injury or serious physical pain to an animal." Scientific
research is exempt "unless the risk of injury or serious physical pain is disproportionate
to the benefit expected from the research." Although tabled in Canada's House of
Commons May 19, 1988 (Anon., 1988c), no action has been taken to date to implement
the report.

In 1989, the CFHS proposed federal legislation governing experimental animals, which
included economic sanctions and more power for non-user members of institutional
ACCs (Anon., 1989). The CFHS proposal was later refused by the then Minister of
National Health and Welfare, The Hon. Perrin Beatty, who considered it would be more
expensive than CCAC's program, and "would not improve the already excellent level of
care given to animals in Canadian laboratories."

The federal Health of Animals Act, C-66 (June, 1990, rev. March, 1992); 38-39
Elizabeth II, Chapter 21 (replacing the Animal Disease and Protection Act and Part III
of the Livestock and Livestock Products Act) is aimed at protecting Canadian livestock

16
from contagious diseases such as tuberculosis and brucellosis, and keeping out foreign
diseases. Under Regulations, the Act states that "the Governor in Council may make
regulations for the purpose of protecting human and animal health...including
regulations...

i) for the humane treatment of animals and generally governing the care, handling and
disposition of animals;

ii) governing the manner in which animals are transported within, into or out of
Canada; and

iii) providing for the treatment or disposal of animals that are not cared for, handled or
transported in a humane manner."

Agriculture Canada has published Codes of Practice for pigs, veal calves, poultry, dairy
cattle, beef cattle, ranched fox and mink (Agriculture Canada, 1771/E, 1984; 1821/E,
1988; 1757/E, 1989; 1853/E, 1990; 1870/E, 1992; 1831/E, 1989; 1819/E, 1988). In
addition, a revision of the Recommended Code of Practice for the Care and Handling of
Farm Animals--Pigs (1898/E) is now in press. The Codes represent the current industry
standards and, for the most part, are the minimum CCAC requires for research
institutions undertaking agricultural research. Researchers and others working with
agricultural animals must be fully conversant with these Codes.

The federal Food and Drug Act and Regulations (August 5, 1982), cover use of animals
in the testing of new drugs and vaccines, and for toxins in foods. Health and Welfare
Canada's Bureau of Biologics is responsible for the monitoring of biologics, the virulence
and efficacy of which can usually be tested only in the living animal.

Very little testing of cosmetics and consumer chemicals is conducted in Canada due to the
fact that almost none of these items are developed here (Gilman, 1980). That said,
cosmetics are the responsibility of the Drugs Directorate, through the Bureau of Non-
prescription Drugs. Safety evidence is required of the manufacturer and could involve in
vivo (animal) testing in the case of new chemicals and chemical combinations.

Responsibility for food safety/nutritional quality lies with the Bureau of Microbiological
Hazards, Chemical Safety, and Nutrition. The identification of specific microorganisms
and toxins in food requires some specific animal tests (e.g., tests are conducted in mice
for paralytic shellfish toxins and botulism).

b) Provincial Legislation

i) Saskatchewan

Under the Veterinarians Act of 1987 of the Province of Saskatchewan (Chapter V-5.1) "a
person using an animal in research at a university," and employing procedures in studies
approved by an ACC which includes a veterinarian, is exempt from the Act's provision

17
that only a member of the Saskatchewan Veterinary Medical Association "shall
engage...in the practice of veterinary medicine."

ii) Alberta

Alberta, in 1966, passed its Universities Act, (Section 50, "Dog Control and
Procurement"; Regs. 341-366). This legislation forbade research institutions from
purchasing dogs for research purposes, but made it incumbent upon municipal pounds to
make all unclaimed dogs available to faculties of medicine on request. In 1972, Alberta
Regulation 33-72 was expanded to cover the treatment of animals. Conditions under
which they were to be transported, maintained, used and disposed of were specified. This
included mandatory exercise for caged dogs. It required use of anesthesia, analgesia, and
standards of post-surgical treatment; the demand that those using animals be qualified;
and safeguards to permit owners' retrieval of their animals from an isolation/quarantine
facility (Secord, 1974).

iii) Ontario

The use of experimental animals in Ontario is governed by its Animals for Research Act
(Revised Statutes of Ontario, 1980, Chapter 22 as amended by 1989, Chapter 72,s6 and
Regulations 16,17,18,19. Revised Regulations of Ontario, 1980, March 1990). Before its
passage after much controversy in May, 1971, the then Minister of Agriculture and Food,
The Hon. William Stewart, stated February 19, 1969 in the Provincial Legislature, that:
"By controlling the source of animals and by making animals more readily available to
research facilities, undesirable practices such as the theft of dogs--dognapping--and the
operations of unscrupulous dealers will be curbed..." (Stewart, 1969a). On June 17, 1969,
again in the Provincial Legislature, Mr. Stewart said that: "When this legislation is
enacted, the Government of Ontario will embody in these Regulations the principles
relating to the care of experimental animals as spelled out by the Canadian Council on
Animal Care (CCAC)..." (Stewart, 1969b); however, this was never done.

This Act, which is administered by the Ontario Ministry of Agriculture and Food
(OMAF), requires annual registration of all research facilities in the province. It includes
clauses requiring anesthetics and analgesics in order to prevent animal suffering and
unnecessary pain, local ACCs which include a veterinarian and authority to require that
research projects be modified; regulated minimum standards for housing, procedures and
care, inspection of research premises and a minimum redemption period of 72 hours
(which municipalities may extend). Under the Act, after this period, the pound may sell
unclaimed animals for pets, hunting and for use in registered research establishments.

Under the Animals for Research Act, the Ontario Society for the Prevention of Cruelty
to Animals Act, 1955 "does not apply in respect of animals in the possession of the
operator of a registered research facility (RSO 1980, c.22, s.19)."

In 1985, Bill 21, a Private Member's Bill proposed by MLA Ed Philip, would have
allowed municipalities the choice of whether or not to provide unwanted stray animals to

18
research establishments. In 1986, according to the Ontario Public Trustee's Office, the
Coalition Against Pound Seizure, "a project funded by the Toronto Humane Society"
spent more than $200,000 fighting the Animals for Research Act (McAndrew, 1987). Bill
21 was opposed by veterinary societies (Sanderson, 1986), and the Ontario Humane
Society, which suggested that its members disassociate themselves from the coalition.
Researchers and academics argued that work crucial to saving human lives was
threatened because of pressure from such groups (Wilson, 1986).

Although Bill 21 passed 2nd Reading in 1985, it was not put forth for 3rd (and final)
Reading (Anon., 1987; Comeau, 1987; Sheppard, 1987; Wilson, 1986).

In 1988, MLA C.J. "Bud" Wildman introduced Bill 190, which sought to amend the
Animals for Research Act by prohibiting use of animals in non-medical testing. Hearings
were held to debate it in 1989 (Anon., 1989; Harvey, 1990) and it received 2nd Reading,
but not the necessary 3rd Reading. However, meetings in 1991/92 between Ontario's
Minister of Agriculture and Food, The Hon. Elmer Buchanan, and various groups have
resulted in his decision to "increase public input into the research process as well as
restrict the usage of animals in testing."

iv) Quebec

Following charges that 2,000 stray dogs were picked up off suburban Montreal streets
and sold in the U.S. for use in research (CP, 1985), the Montreal-based Canadian SPCA
conducted an investigation which found that 3,000-5,000 dogs and 2,000 to 3,000 cats
were being shipped each year from Quebec, primarily because research establishments in
the Northeastern U.S. had passed laws against the sale of animals from municipal pounds
for research (Duquette, 1986). The CSPCA subsequently developed and proposed
legislation governing animal use (Duquette, 1986).

More recently, a provincial Table de concertation (Working Group) on Cruelty to Animals

examined animal welfare in the Province of Quebec, and reviewed animal welfare
legislation proposed by the CSPCA (Anon., 1991a). The Table's (1992) report requested
that CCAC ensure strict adherence to its guidelines in Quebec by increasing the number
of assessments, and unannounced visits in particular, ensure effective functioning of
ACCs, pay particular attention to the qualifications of researchers and animal care
attendants, ensure management and monitoring are adequate, strongly encourage
adherence to Russell and Burch's "3R" tenet of Replacement, Reduction and Refinement
(Russell and Burch, 1959) and ensure ACCs include representatives from the community,
including known animal welfare societies.

The recommendations also asked Quebec ministries involved in animal use to ensure
CCAC standards were applied in all institutions receiving provincial funding, and that
CCAC periodically supply the Ministry of Agriculture, Fisheries and Food with a list of
the institutions assessed, so that the Ministry could request copies of assessment reports.

c) Legislation in the United Kingdom

19
The U.K.'s Animals (Scientific Procedures) Act 1986, which replaced the century-old
Cruelty to Animals Act 1876, gives the Home Secretary the responsibility for judging the
scientific merit of the research he authorizes, "for which he will be answerable to
parliament" (Hollands, 1986). Section 5(4) states that "In determining whether and on
what terms to grant a project licence, the Secretary of State shall weigh the likely adverse
effects on the animals concerned against the benefit likely to accrue as a result of the
program to be specified in the licence." This clause was included at the insistence of an
alliance comprising the British Veterinary Association (BVA), the Committee for the
Reform of Animal Experimentation (CRAE), and the Fund for the Replacement of
Animals in Medical Experiments (FRAME) (Smith, 1988).

Commenting on this requirement, Balls (1989) states: "What then will follow is the
crucial weighing of the balance, but how certain will we need to be that a particular
program of work will lead to the relief or avoidance of significant human (or animal)
suffering before we allow undeniably painful procedures to be applied to laboratory
animals? One part of the answer should be to apply a more rigorous and more considerate
and humane analysis in the future than in the past, where (to agree with Les Brown)
(Brown, 1988) supposed benefits couched in generalities such as the 'increase of
knowledge' have all too often been enough. Also, let us in future be more willing to give
the animal the benefit of the doubt--for who can have the foresight to satisfactorily and
convincingly quantify the extent to which the rate of progress in medical research would
really suffer as a consequence?"

The Act also calls for a statutory 21-member Animal Procedures Committee (APC) which
has wide powers to advise the Home Secretary, who controls the overall severity
permitted. It also dictates that all research animals (with some exceptions (e.g., farm
animals and animals taken from the wild) be obtained from registered suppliers (Balls,
1986). The APC was to provide the mechanism for ethical debates. However, member
Judith Hampson, former Animal Experiments Officer with the RSPCA, has stated that the
committee deals only with a few cases referred to it by the Home Office and "has failed
to deal adequately with issues of particular concern to the public...and there is very little
public accountability." She called for formation of institutional ethics committees with
community representation (Hampson, 1992).

The Animals (Scientific Procedures) Act was widely debated and criticized, both before
and after passage (Balls, 1990; Aldhous, 1990; McKie, 1986; Fisher, 1990), and was even
described by animal activists as "the vivisectors' guide to pain" (Churchward, 1986).
Others subsequently questioned whether the Act had failed (Balls, 1990; Anon., 1990)
when experiments carried out in an inadequately anesthetized rabbit by an elderly
neurophysiologist at the National Institute for Medical Research (NIMR) were exposed
by an antivivisectionist group (Anon., 1990) using videotaped evidence. The organization
later claimed that "the Home Secretary failed to weigh adequately the likely benefit of the
research against the adverse effects on the animals involved."

The scientist and his assistant relinquished their licences and a subsequent Medical

20
Research Council (MRC) enquiry reported that the requirements of the Act had, indeed,
been broken (Anon., 1991b). Many felt, however, that the case was an isolated incident,
and the British Veterinary Association (BVA) came to the conclusion that, overall, the Act
was working well, although improvements are needed in administration and operation
(Anon., 1991c).

d) Legislation in the United States

In the U.S., two primary federal laws govern animal use. The Health Research
Extension Act (or NIH Authorization Act) passed in 1985 requires establishment of
Institutional Animal Care Committees (IACCs) and IACC inspections (Traystman, 1990).
The Animal Welfare Act (1985) includes in its requirements provision of adequate
veterinary care with appropriate use of anesthetic, analgesic, tranquillizing drugs, or
euthanasia, consideration by the principal investigator of alternatives to any procedure
likely to produce pain and distress, and establishment of ACCs, which must include a
veterinarian.

The Department of Agriculture, Animal and Plant Health Inspection Service, March 15,
1989 published Animal Welfare; Proposed Rules in the Federal Register (pgs. 33447-
33531) to amend the Animal Welfare Act (7 U.S.C. 2131-2157), which itself was an
amendment to the Farm Bill (1985). The 1985 amendments had required standards for
pre- and post-surgical care, inspection of research facilities, requirement for ACCs,
establishment of an information service at the National Agricultural Library, annual
training sessions for laboratory personnel and increased penalties for facilities violating
the animal welfare standards (Schwindaman, 1990).

After much public input and debate (Meyers, 1990), Final Rules for Part One
(definitions) and Part Two (regulations) were published August 31, 1989 in the Federal
Register (pgs. 36112-36163). Although these Final Rules approved the governing of
small animals, Part Three proposed "additional standards...directed primarily toward the
exercise of dogs, the psychological well-being of primates and standards for research
facilities" (Schwindaman, 1990).

Eventually, the Final Regulations, issued February 15, 1991, and published in the
Federal Register (pgs. 6426-6505) were "performance based," i.e., the health of the
animals would be of greater importance than details of their housing (Anon., 1991c). By
August 14, 1991, institutions had to produce written plans for providing exercise for dogs
and improving the psychological well-being of non-human primates (NHP) (Myers,
1991). However, a U.S. judge has invalidated the regulations because they did not set
minimum standards (Mervis, 1993).

As the result of a lawsuit launched by the Humane Society of the United States (HSUS)
and the Animal Legal Defence Fund (ALDF), experimental mice, rats and fish will now
be covered by the legislation in the U.S. January 8, 1992, the U.S. district court in
Washington, D.C. ruled that the U.S. Department of Agriculture (USDA) had violated the
federal Animal Welfare Act by denying basic protection to America's 15 million mice,

21
rats and birds used annually in research (Anon., 1992a). However, the government has
appealed this decision (Mervis, 1993). Institutional Animal Care and Use Committees
(IACUC) (Orlans, Simmonds and Dodds, 1987), must now review all research proposals
involving these species, as the CCAC has required since its inception.

A compilation of U.S. state laws concerning research animal use has recently been
published (NABR, 1991), as well as animal-related legislation introduced in the 102nd
Congress (Anon., 1992b). August 26, 1992, the U.S. also passed the "Animal Enterprise
Protection Act of 1992" (Public Law 102-346) which made it a Federal offence to enter a
research laboratory without authorization and "steal, destroy or make unauthorized use of
research animals, equipment or data" (Heflin, 1992; Anon., 1992c).

6. Pre-University Use of Animals

Before the establishment of CCAC guidelines, pre-university use of experimental animals

was governed by a one-page document prepared by the CFBS. The Federation required
compliance with its Guiding Principles, and said that "all experiments employing animals
must be carried out under the supervision of a qualified teacher."

At the present time, animal use in the school is subject to the requirements of legislation
such as the Health of Animals Act (Bill C-66), the Criminal Code of Canada, Section 446,
Cruelty to Animals, and provincial legislation, where such exists. Primary responsibility
for animal use at the pre-university level now lies, however, with the Youth Science
Foundation (YSF) (904-151 Slater St., Ottawa, Ontario Canada K1P 5H3), which
requires compliance with the CCAC guidelines in the conduct of biological research.

The YSF, amongst its responsibilities, regulates animal experimentation in Science Fairs.
All research intended for Science Fairs must be screened by a committee cognizant of
current requirements; if none is available, the YSF may be contacted. Science Fair
Regulations permit use of lower forms of life (bacteria, fungi, protozoa, insects, plants
and invertebrate animals). Vertebrate animals (birds, fish, mammals, reptiles, amphibians)
"are not to be used in any active experiments which may be deleterious to the health,
comfort or physical integrity of the animal..." Observation of wild animals, animals in
zoological parks, farm animals and pets is permitted.

It should be noted that, before any such projects are undertaken, adequate arrangements
should be made for the care of the animal while in the classroom, and its subsequent
disposition, which may involve euthanasia.

Some school boards, such as the Peel Board of Education, have produced their own
guidelines (Henshall, Scott and Scott, 1986). The Ontario Egg Producers' Marketing
Board (7195 Millcreek Dr., Mississauga, Ontario Canada L5N 4H1) has also published A
Teacher's Guide to Hatching Eggs in the Classroom (1990). An American leaders' manual
for avian embryology, entitled Beginning of Life, is also available (Publ. #408-029.
Virginia Co-operative Extension Service; Clinton, V. Turner, Administrator, 1890
Extension Program, Virginia State University, Petersburg, VA).

22
Guidelines have also been published in the U.S. (NABT, 1990; Orlans, 1977; McGiffin
and Brownley, 1980; ILAR, 1989), and in the U.K. by the RSPCA (1985) (Causeway,
Horsham, West Sussex RH12 1HG, U.K.).

B. LOCAL LEVEL

1. The Institutional Animal Care Committee

It is essential that the necessity for and the benefits of effective control in the care and use
of experimental animals be recognized. Regardless of whether this control is "voluntary"
or legislated, each institution has a commitment to be cognizant of the nature of all
experiments involving animals in their establishments and to ensure their propriety. This
responsibility is best met by an effective local ACC, reporting to the appropriate senior
administrative officer of the institution. The local ACC should be responsible for
formulating and implementing policy on all matters concerning the general care and use
of animals as outlined below.

Terms of Reference for Animal Care Committees* (Revised in 2006)

1. Membership | 2. Authority | 3. Responsibility | 4. Meetings | 5. General

The Canadian Council on Animal Care (CCAC) requires that institutions conducting
animal based research, teaching or testing establish an animal care committee (ACC), and
that it be functionally active. Each committee's operation must be governed by formal
Terms of Reference that include the following Terms, but need not be limited to them.
The ACC's Terms of Reference must be tailored to reflect and refer to the institution's
animal care and use program, including the members of the program and the institution's
policies, practices and procedures.

Most institutions have a single ACC. A few large institutions choose to have more than
one ACC: this is acceptable, as long as the ACC system is well structured to avoid
potential conflicts of interest, and has an institutional ACC that oversees the work of the
ACCs for the various units and establishes policies and procedures to ensure sound
general standards in order to meet CCAC guidelines throughout the institution. The
elements covered in this policy statement may be divided out between the various ACCs
of an institution, as long as all elements are covered in an appropriate and structured
fashion and are defined in suitable terms of reference for each committee.

ACCs may also choose to form subcommittees to work on specific areas such as protocol
review or development of standard operating procedures (SOPs). Protocol review

23
subcommittees should include at least one scientist, one veterinarian, one community
representative, one institutional member who does not use animals, one technical staff
representative and the ACC coordinator.

Institutional ACCs should be responsible directly to the senior administrator responsible

for animal care and use for the institution (president, vice president, rector, CEO, etc.),
and this link should be specified in writing. ACCs for faculties or other divisions of the
institution should have representation on the institutional committee and report directly to
it, in addition to the reporting line(s) to any other senior administrators. The CCAC policy
statement on: senior administrators of animal care and use programs (in preparation)
should be consulted for details on the roles and responsibilities of the institution and its
senior administrators.

The institution must work with the ACC to ensure that all animal users and caregivers are
informed of and comply with institutional animal care and use policies and procedures.

The institution must be supportive of the committee's work. This includes appointing an
ACC coordinator, who may work part-time for the ACC in the case of smaller
institutions, whereas larger institutions will need one or more employee(s) to accomplish
this work. The ACC coordinator must support the ACC by ensuring that animal use
protocols are well managed, that committee minutes and reports are promptly produced
and distributed, that all exchanges between the ACC and animal users are well
documented and filed in a timely manner, and that animal users and ACC members are
provided with necessary information.

The institution must also ensure that ACC members are provided with training
opportunities to understand their work and role: these must include at least a formal
orientation session, to introduce new ACC members to the institution's animal care and
use program and its members, policies and procedures, as well as to the animal facilities
and to CCAC guidelines and policies. Material on the CCAC website (and other relevant
websites), such as the Modules on the Core Topics of the Laboratory Animal/Teaching
Stream of the CCAC Recommended Syllabus, can be introduced as possible resources.
Ongoing opportunities to better understand animal care and use in science should also be
provided, such as time spent with animal care givers and users, access to relevant journals
and materials, and meetings/workshops related to animal care and use, including the
CCAC National Workshop.

The institution and its senior administrators must also ensure that the ACC is well
respected within the institution, and that all ACC members and the ACC Chair are valued
and recognized.

* May also be referred to as:

institutional animal care and use

committees (IACUC)
animal research ethics boards

24
 (AREB)
Ethics Committees

and by other names, as long as their function

is clear and they operate according to Terms
of
Reference based on this document.

1. Membership

ACC members should be appointed for terms of no less than two years and no more than
four years, renewable only up to a maximum of eight consecutive years of service. This
maximum should not be exceeded, except in the case of very small institutions (i.e. those
that have 3 or fewer animal users). This does not apply to ACC members who must be
part of the ACC because of their role within the institution (ex officio members): the ACC
Coordinator, the veterinarian(s) and the animal facility manager. The complement of the
committees will vary and should be determined by the needs of each institution, but
should include:

a) scientists and/or teachers experienced in animal care and use, who may or may not be
actively using animals during their term on the ACC; there should be a minimum of
two such members, and representation of all the major animal-using divisions of the
institution must be ensured;
b)
a veterinarian, normally experienced in experimental animal care and use;
an institutional member whose normal activities, past or present, do not depend on or
c) involve animal use for research, teaching or testing;
at least one, and preferably two or more, person(s) representing community interests
d) and concerns, who has (have) had no affiliation with the institution, and who has
(have) not been involved in animal use for research, teaching or testing; community
representation must be ensured for all ACC activities throughout the year;
technical staff representation (either an animal care, an animal facility or an animal
e) research technician) if there is (are) (a) technical staff member(s) actively involved in
animal care and/or use within the institution;
student representation (graduate and/or undergraduate), in the case of institutions that
f) have programs where students use animals; and
the ACC coordinator (the institutional employee who provides support to the ACC).
g)

The senior administrator to whom the committee reports must not be a member of the
ACC, but there can be a representative of the senior administration on the committee.

25
The person with overall responsibility for the animal facilities, whether a veterinarian, a
scientist or a technical staff member, must be included on the ACC. In large institutions
with several facilities, consideration can be given to having individual facility managers
included on the ACC on a rotating basis.

ACCs benefit from having occupational health and safety and biosafety representatives
(if this is not the case, other ways must be found of ensuring close links), and ACCs can
also benefit from the presence of biostatisticians, ethicists and those responsible for
public relations.

Every ACC must have a chair who should not be directly involved in the management of
the institutional animal facilities, nor be a clinical veterinarian for the institution, nor be
an animal health or veterinary personnel member charged with ensuring compliance with
CCAC guidelines, nor be involved in the preparation of a significant number of the
protocols to be reviewed by the committee, in order to avoid potential conflicts of
interest. Provision should be made to co-opt other persons to the ACC as the need arises.
A reasonable quorum, such as a majority of the members, should be established for ACC
meetings, and the quorum should include community and veterinary representation.
Meetings should be scheduled at times that are convenient for all members, including
community representatives.

2. Authority

The ACC must have the authority, on behalf of the senior administrator responsible for
animal care and use for the institution, to:

a) Stop any objectionable procedure if it considers that unnecessary distress or pain is

being experienced by an animal;
b) Stop immediately any use of animals which deviates from the approved use, any non
approved procedure, or any procedure causing unforeseen pain or distress to animals;
and
c) Have an animal killed humanely if pain or distress caused to the animal is not part of
the approved protocol and cannot be alleviated.

The Chair of the ACC and the veterinarian(s) must have access at all times to all areas
where animals are or may be held or used.

Each institution must establish procedures for post-approval monitoring of animal use
protocols, and must define the roles and responsibilities of the members of the animal
care and use program in the monitoring process. The institutional ACC is the body
responsible for determining and working to correct breaches of compliance with
approved animal use protocols and SOPs. Breaches of compliance that cannot be
corrected by the ACC working with the concerned animal users and veterinary/animal
care staff must be referred to the senior administration, which must inform all members

26
of the animal care and use program about sanctions that will be taken by the
administration in the event of serious breaches of compliance.

As the ACC is generally not present when animal use protocols are being undertaken, the
committee must work with the members of the veterinary and animal care staff to ensure
compliance with its decisions and with the conditions set out in approved protocols. The
veterinary and animal care staff must work in a collegial manner with animal users and
attempt to correct deficiencies collaboratively. Where there are persistent breaches of
compliance or threats to the health and safety of personnel or animals, these must be
reported back to the Chair of the ACC, and the Chair and ACC must promptly address
these issues, through communications with the animal user(s), meetings and site visits,
and eventually communications with the senior administrator, as necessary.

The ACC must also delegate to the veterinarian(s) the authority to treat, remove from a
study or euthanize, if necessary, an animal according to the veterinarian's professional
judgment. The veterinarian must attempt to contact the animal user whose animal is in
poor condition before beginning any treatment that has not previously been agreed upon,
and must also attempt to contact the ACC Chair, but the veterinarian must have the
authority to proceed with any necessary emergency measures, whether or not the animal
user and ACC Chair are available. A written report should be sent by the veterinarian to
the animal user and to the ACC following any such event.

The veterinarian and ACC may also choose to delegate certain responsibilities to one or
more senior animal care staff member(s).

3. Responsibility

It is the responsibility of the ACC to:

a) Ensure that no research or testing project or teaching program (including field

studies) involving animals be commenced without prior ACC approval of a written
use protocol; further to this, that no animals be acquired or used before such
approval. This includes internally funded projects;
b) Ensure that no animals be held for display or breeding purposes, or for eventual use
in research, teaching or testing projects, without prior ACC approval of a written
animal use protocol, except where current CCAC guidelines provide for exemptions.
The ACC should also be aware of other animal-based activities, such as commercial
or recreational activities, within the institution, and should work with the persons
responsible for these activities to ensure that animal care and use is undertaken
according to appropriate procedures;
c) Require all animal users to complete an animal use protocol form and ensure that the
information therein includes the following points, clearly presented in a form that all
members of the ACC can readily understand (supplemental information can be found
in the CCAC guidelines on: animal use protocol review, 1997). To facilitate the work
of both protocol authors and ACC members, appropriate SOPs should be referred to

27
as much as possible. Approved protocols and SOPs should be readily available in the
areas where animal-based work is taking place.
i) project title and descriptive procedural keywords or brief description of the
procedures to be conducted on animals, as defined in the CCAC Animal Use
Data Form;
ii) principal investigators/teachers, and all personnel (post-doctoral fellows,
research staff, graduate and undergraduate students) who will handle animals,
along with their training and qualifications with respect to animal handling (see
point 3m) iii)); in the case of undergraduate students, who may have very little
training, close supervision is required;
iii) departmental affiliation;
iv) proposed start date, proposed end date (if the study is to take place over more
than one year, the work and numbers of animals for the first year only should
be approved, and further work can then be approved in yearly protocol
renewal(s) or new protocols - see Section 3g) on protocol renewals);
v) for research or testing projects, funding source(s) and status of funding
approval;
vi) for research projects, an indication of whether the project has received peer
review for scientific merit;
vii) for teaching programs, a course number and an indication of whether the course
has been re-viewed with respect to the pedagogical merit of using live animals;
institutional or departmental curriculum committees can be called upon to
provide a review of pedagogical merit to the ACC; a specific appendix or
separate protocol form can be used to better capture information relevant to the
ethical review of teaching programs (see Section 12 of the CCAC guidelines
on: animal use protocol review);
viii) for testing projects, an indication that the testing has been planned according to
the most current regulatory requirements, using guidelines acceptable to the
regulatory agency(ies) and which meet the requirements of the CCAC policy
statement on: ethics of animal investigation; that the planned animal use not
exceed the requirements of the regulatory authorities - if it does, justification
for the additional animal use must be provided;
ix) lay summary;
x) an indication of the use of biohazardous, infectious, biological, chemical or
radioactive agents in animal-based projects; and, if so, an indication of
institutional approval of this use;
xi) category(ies) of invasiveness as defined in the CCAC policy statement on:
categories of invasiveness in animal experiments, and Purpose of Animal Use
(PAU) as defined in the CCAC Animal Use Data Form;

28
xii) information with regard to the Three Rs (replacement, reduction and
refinement alternatives) of animal use, to include:
xii.1 a description of why sentient animals must be used for the project, of
how the applicant arrived at this conclusion (e.g., searches of databases
on alternatives), and of possible replacement alternatives (non-animal
methods, cell/tissue culture, computer simulations, audio-visual teaching
methods, the replacement of sentient animals with animals of lower
sentiency, etc.) and justification if these are not to be employed;
xii.2 justification of the species and numbers of animals to be used over the
course of the year, to emphasize reduction of animal use within an
appropriate experimental design, while ensuring that sufficient numbers
of animals will be used to fulfill requirements for statistical
significance/scientific validity in the case of research projects, or for
acceptance of regulatory tests;
xii.3 a description of all of the refinements to be employed to protect and
enhance animal health and welfare, which may include:
xii.3.1 anesthesia and analgesia, including dosages and methods of use,
for all invasive protocols; strong scientific justification must be
provided for not using anesthesia or analgesia in the case of
invasive protocols;
xii.3.2 other medical treatments as appropriate, as indicated through
veterinary consultations;
xii.3.3 housing and husbandry methods, and environmental enrichment
as a means to refine animal care; any limitations on
environmental enrichment from that normally offered to animals
in the institution, based on CCAC guidance, must be justified to
the ACC;
xii.3.4 refinements to the procedures to be employed on the animals;
xii.3.5 refinements to the length of time that animals will be held/used;
xii.3.6 any other possible refinements;
xiii) a clear description detailing the procedures that are carried out on the
animals (referring to appropriate SOPs as much as possible); the use of
graphic representations is encouraged;
xiv) a description of the endpoint(s) of the experimentation, selected
according to the CCAC guidelines on: choosing an appropriate
endpoint in experiments using animals for research, teaching and
testing, 1998 (refer to institutional SOPs, if available and relevant); the
person(s) responsible for monitoring the animals and applying endpoints
should be identified, and the schedule for monitoring animals and any
relevant checklists of signs and symptoms to be used when evaluating

29
 the animals should be included; all protocols, even non-invasive ones,
must identify endpoints, to ensure that any animals requiring treatment
are treated and that animals are not simply kept indefinitely; relevant
information for identifying and applying endpoints must be readily
available, preferably posted, in the area where the animal-based work is
taking place;
xv) a description of capture, restraint, transportation and/or housing of
animals used in field studies, as well as any other information pertinent
to field studies, such as capture of non target species, ecological impacts
and potential injuries or mortality during capture or transportation, if
relevant; wildlife studies should be addressed in either a separate section
or appendix of the protocol form, or can have their own protocol form,
especially where a significant number of wildlife studies are undertaken
(see the suggested wildlife protocol form in Appendix B of the 2003
CCAC guidelines on: the care and use of wildlife);
xvi) the method of euthanasia, if used; justification for any physical
euthanasia methods, or for any methods that deviate from those
described in the most recent CCAC guidance on euthanasia;
xvii) a description of the fate of the animals if they are not to be euthanized,
including the length of time that they are to be held;
xviii) any other information considered important or necessary and pertinent,
including information or results derived from any relevant previous
protocols; the description and use of previous relevant results is
particularly important to ensure that methodologies are not simply re-
used without learning from any animal welfare problems that were
encountered in the past, that the protocol continues to have relevant
goals and methodology, and that appropriate refinements to protect and
enhance animal welfare are sought and implemented;
d) Ensure that each research project has been found to have scientific merit through
independent peer review before approving the project; if the review is not carried out
by an external, peer review agency, the institution should require that it be obtained
according to the CCAC policy statement on: the importance of independent peer
review of the scientific merit of animal based research projects, 2000. The institution
must implement a mechanism through which non-peer-reviewed projects are
reviewed for their scientific merit either by calling upon the expertise of individual
independent peers or by making use of scientific committees or advisory boards;
e) Review and assess all animal use protocols, with particular emphasis on the CCAC
policy statement on: ethics of animal investigation and CCAC guidelines on: animal
use protocol review as well as on all other relevant CCAC guidelines and policy
statements and, where necessary, require further supportive information from the
investigator/teacher or meet with the investigator/teacher to ensure that all members
of the committee understand the procedures to be used on the animal. Information
exchanges and ACC discussions with protocol authors can be very useful, but

30
 protocol authors and members of their teams must always clearly remove themselves
from ACC decision-making on their own protocols.

The committee must also ensure that all procedures comply with CCAC guidelines,
and, if at variance with those guidelines, require justification for the variance on
scientific grounds. ACCs should both discuss protocols and make decisions on them
during full committee meetings, rather than through individual reviews, and should
attempt to reach decisions by consensus. Electronic tools are widely used for
protocol management purposes and to facilitate and expedite the submission and
review of protocols. This is encouraged as long as ACCs or protocol review
subcommittees continue to meet in person for protocol discussions and final
approvals.

An ACC may delegate the responsibility of interim approvals to an interim approval

subcommittee, which must include at least one scientific member, one veterinarian
and one community representative, one of which should preferably be the chair of
the ACC. However, such interim approvals should only be used infrequently, and the
interim review process, including exchanges between the ACC and protocol authors,
must be documented and must then be subject to discussion and final approval at a
full meeting of the committee. The ACC should define its own protocol review
process, with or without (a) protocol review subcommittee(s), in its Terms of
Reference. This process should include or refer to clear instructions to protocol
authors, to ensure that all animal users in the institution understand how the ACC
works, when it meets, how to fill out and submit a protocol form and what to expect
after submission of the form;
f) Ensure that animal users update their protocols with any modifications they intend to
make, and approve any modifications to a protocol before they are implemented.
Minor modifications (e.g., 1 or 2 animal users added or removed, a small number of
animals added, etc.), as defined by the ACC, can be approved by the Chair of the
ACC or a delegate.

For any major changes to a protocol, require that a new one be submitted. ACCs
should define, in writing, their own criteria as to what constitutes a major change to a
protocol (e.g., a considerable increase of the number of animals required vs. the
number in the original protocol, a change of species, use of more invasive or more
frequent procedures, use of entirely new procedures, or other criteria).

Ensure that animal users report any unanticipated problems or complications, as well
as on the steps they have taken to address the problem(s), to the ACC;
g) Review all protocols annually, i.e., within a year of commencement of the project;
annual renewals should be approved by at least a scientist, a veterinarian and a
community representative and should be brought to the attention of the full ACC for
its information. Institutions may choose to use a shorter protocol renewal form, but
no matter what form is used, all protocol renewals must emphasize:

31
 i) the number of animals used in the preceding year;
ii) the number of animals needed for the year to come, with a justification;
iii) a brief progress report, describing any complications encountered relative to
animal use (unpredicted outcomes, and any animal pain, distress or mortality),
any amendments to the original protocol, and any progress made with respect
to the Three Rs of replacement, reduction and refinement of animal use;
iv) a brief report on the adequacy of the endpoints for the protocol, and on any
complications encountered or refinements made relative to protecting animals
from pain, distress or mortality; and
v)
any other changes from the original protocol.
Require the submission of a new protocol after a maximum of three consecutive
renewals;
h) Document all ACC discussions and decisions in the committee minutes and on
attachments to the protocol forms;
i) Define an institutional appeal mechanism that can be used by the author of a protocol
in the event that animal use is not approved by the ACC. This mechanism should
include appropriate expertise and ensure a separate, fair and impartial process. The
CCAC may be called upon for information purposes; however, appeals cannot be
directed to the CCAC;
j) Ensure that all ACC members and animal users have the opportunity to become
familiar with the CCAC Guide and CCAC policy statement on: ethics of animal
investigation and all other CCAC guidelines and policy statements, federal,
provincial or municipal statutes that may apply, as well as institutional requirements;
k) Ensure appropriate care of animals in all stages of their life and in all experimental
situations. Veterinary care must be available. Formal arrangements must be made to
obtain the services of a veterinarian, at least on a consultative basis, if they are not
readily available within the institution. These formal arrangements must be based on
the elements contained in the CALAM/ACMAL Standards of Veterinary Care of the
Canadian Association for Laboratory Animal Medicine (2004), which define the
roles and responsibilities of veterinarians involved in scientific animal care and use
programs;
l) Establish procedures, commensurate with current veterinary standards, to ensure that:
i) unnecessary pain or distress is avoided, and animal stress and injuries are
avoided, whether during transfers of animals or in their normal quarters;
ii) anesthesia and analgesia are properly and effectively used; the only exception
to this may be when agents must be withheld as a scientifically justified
requirement of the study, and that this has been approved by the ACC. Painful
studies requiring exemption from the use of either anesthetics or analgesia must

32
 be subject to particular scrutiny, not only prior to approval, but also during the
experiment;
iii) appropriate post-operative care is provided;
iv) all due consideration is given to animal welfare, including environmental
enrichment;
m) Ensure that policies to provide for a system of animal care that will meet the needs of
the institution are established and implemented, and include:
i) the requirement that all animal care and animal experimentation are conducted
according to CCAC guidelines and policies, and to any federal, provincial and
institutional regulations that may be in effect;
ii) ensuring adequate animal care and management of the animal facilities, in
particular by verifying that there is a person clearly designated to be in charge
of animal care and management of the animal facilities, who should be a
member of the ACC (see Section 1), and who should keep the other ACC
members updated on the activities within the animal facilities;
iii) the training and qualifications of animal users and animal care personnel;
veterinarians and animal care staff must receive continuing education in their
field, and animal users (scientists/study directors, post-doctoral fellows,
graduate students and research technicians) must receive appropriate training
according to the CCAC guidelines on: institutional animal user training, 1999,
either within the institution or through the programs of other institutions;
iv) an occupational health and safety program for those involved in animal care
and use, in collaboration with the institutional authorities on occupational
health and safety, that will appropriately protect all those who may be affected
by animal-based work, according to CCAC guidelines (see Chapter VIII of
Volume 1 (2nd Edn, 1993) of the CCAC Guide or the most recent CCAC
guidance on occupational health and safety);
v) standards of husbandry, facilities and equipment;
vi) standard operating procedures for all activities and procedures that involve
animals, including animal care and facility management SOPs (typically
produced by the veterinary and animal care staff), and animal use SOPs
(typically produced by animal users, in collaboration with veterinary/animal
care staff as needed); the ACC should receive all SOPs and ensure that all
necessary SOPs are produced and regularly reviewed (see also Section 5a)iii));
vii) procedures for euthanasia;
n) Encourage the use of pilot studies with few animals when new approaches, methods
or products are being tried, before approving new, large scale protocols. Ensure that
animal users report on the results of any pilot studies, no matter whether they wish to
pursue the study immediately or not, in order to preserve important data on various
approaches to animal-based studies, whether they work well or not; and

33
o) In the case of projects involving proprietary or patentable research or testing, ensure
that as much information as possible is provided to the ACC in terms of what effects
to expect on animal health and welfare, and insist on close monitoring of animals in
order to respect the elements outlined in 3l).

4. Meetings

Animal care committees should meet at least twice per year (most institutions in Canada
have programs which will require more frequent meetings) and as often as necessary to
fulfil their Terms of Reference and be satisfied that all animal use within their jurisdiction
is in compliance with institutional, municipal, federal and provincial regulations, and
CCAC guidelines. Minutes detailing ACC discussions, decisions and modifications to
protocols must be produced for each meeting, and must be forwarded to the senior
administrator responsible for animal care and use.

In addition, the ACC should regularly visit all animal care facilities and areas in which
animals are used, in order to better understand the work being conducted within the
institution, to meet with those working in the animal facilities and animal use areas and
discuss their needs, to monitor animal-based work according to approved protocols and
SOPs, to assess any weaknesses in the facilities (ageing facilities, overcrowding,
insufficient staffing and any other concerns) and to forward any recommendations or
commendations to the person(s) responsible for the facilities and for animal use.

Visits of the animal facilities should be conducted at least once a year, and should be
documented through the ACC minutes or written reports. Those responsible for the
animal facilities should respond to any ACC recommendations in writing, and site visit
reports should always be followed up on jointly by the senior administration and the
ACC. For small institutions, the full ACC may tour the facilities as a group; for larger
institutions, visits to animal care facilities and areas in which animals are used may be
divided between the various members of the committee. No matter what the process
employed, each member of the ACC should participate in some of the facility visit(s) on
an annual basis.

More frequent ACC site visits should be made as necessary to follow up on any protocols
that have raised significant concern during the protocol review process, or where
problems have been encountered with a protocol being carried out in practice or with
other aspects of animal facility operations; these visits may be carried out by the Chair of
the ACC or delegate, accompanied or not by other members or animal care staff.

5. General

The animal care committee:

a) Must regularly review (at least every three years):

i) its Terms of Reference to meet new CCAC guidelines or policies and changing

34
 needs within the institution, the scientific community, the animal welfare
community and society as a whole, and expand its Terms of Reference to meet
the requirements of each institution;
ii) the security of the animals and research facilities;
iii) standard operating procedures and institutional animal care and use policies; SOP
review may be delegated to ACC members with the appropriate expertise, but
SOPs should be accessible to all ACC members, and the full ACC should review
all SOPs that involve procedures that may result in deleterious effects to animal
health or welfare; and
iv) policies and procedures for monitoring animal care and experimental procedures
within the institution, including the identification of the persons responsible for
monitoring animal health and welfare, and the procedures carried out by the ACC
to conduct monitoring;
b) Must maintain liaison with the CCAC Secretariat, and inform the Secretariat of any
changes to their program: to the senior administrator responsible for animal care and
use, the chairperson of the ACC, or the veterinary or senior animal care personnel;
c) Must submit complete and accurate animal use information in the CCAC Animal Use
Data Form (AUDF) format for all protocols annually (animal use information for
each calendar year must be submitted by March 31 of the following year) and also in
pre-assessment documentation;
d) Must develop a crisis management program for the animal facilities and for the
animal care and use program, in conjunction with any general institutional crisis
management plan(s). This program must detail plans in the event of power outages
(short and prolonged), work stoppages, fires, natural disasters, large chemical spills
and other similar crises, and must include a communications plan for addressing
public and media inquiries on concerns related to animal use;
e) Should, from time to time, sponsor seminars or workshops on the use of animals in
science and the ethics of animal experimentation, and encourage as many animal
users, animal caregivers, students, ACC members and other interested parties to
attend as possible;
f) Should try to achieve and maintain a high profile within the institution and in the
community in order to demonstrate the institution's efforts in promoting animal
welfare and to allay some of the public concerns regarding animal experimentation;
and
g) Should be open to developing and maintaining communication with animal welfare
organizations.

March 2006

(previous revision: March 2000)

35
This policy statement supersedes all previous CCAC policies/guidelines on terms of
reference for animal care committees.

2. The Veterinarian

The availability of professional assistance from a veterinarian with interest and

experience in laboratory animal science is of prime importance in achieving and
maintaining optimal conditions of laboratory animal care. In addition to the formal
practice of laboratory animal medicine, the veterinarian should be a major contributor to
the development of each institution's animal care policies and procedures.

"Adequate veterinary care," as defined by the Canadian Association for Laboratory

Animal Medicine (CALAM), [based originally on guidelines prepared by the American
College of Laboratory Animal Medicine (ACLAM)], is the basis upon which CCAC
policy is being developed. It includes: the establishment of Standard Operating
Procedures (SOP) for health monitoring and disease control of laboratory animals; the
prevention of zoonoses; and the responsibility for ensuring that proper precautions are
followed for containment and disease control in specialized colonies such as transgenic
animals, Severe Combined Immune Deficiency (SCID) animals, biohazard studies, and
the personnel working with them.

Veterinarians associated with facilities involved in the practice and evaluation of

livestock intensive management production methods face an especially challenging
situation which includes ensuring that the subject of stress associated with restricted
space, as it may relate to animal behaviour and well-being, is treated objectively (Mench,
Mayer and Krulisch, 1992; Spira, 1986) (see also Social and Behavioural Requirements
of Experimental Animals).

a) General Responsibilities

i) Disease prevention

The veterinarian's duties will include responsibility for a disease prevention program for
all animals maintained within the institution, treatment of ill or injured animals,
maintenance of appropriate health records, provision of advice on anesthesia and
analgesia regimes, antibiotics, anxiolytics, and other therapeutic agents for humane
animal care. The veterinarian should ensure that only approved euthanasia procedures are
conducted and that these are properly carried out (see also Euthanasia). The veterinarian
should also be available for consultation and provide assistance on technical and surgical
procedures.

ii) Education

It will often prove valuable in educational institutions if the veterinarian is involved in the
teaching of surgical principles and other aspects of experimental animal care and

36
handling in courses for senior undergraduates, graduate students, technicians, and
investigators.

iii) Quarantine/conditioning

Serological testing is important in determining and monitoring the epidemiological

characteristics of colony infections (Richter, Lehner and Henrickson, 1984). In addition
to determining if an animal is infected with a disease that may pose a threat to the colony,
the quarantine period also serves as a conditioning period.

A veterinarian or a technician with appropriate training and experience should examine

all animals on their arrival for overt signs of disease which may have been exacerbated
by stress caused by travel (Love, 1980; Reinhardt, 1992). Depending on the species,
source, etc., the quarantine/conditioning period should include routine treatment for
external and internal parasites (Owen, 1992), grooming, cleaning, and providing the
animal with clean food and water (see also Laboratory Animal Care; General Practices).

The veterinarian is responsible for ensuring that animals appearing ill on arrival or
suspected of having been exposed to infection are placed in isolation, examined and
treated. If this is not economically feasible, animals should be humanely euthanized.

Small rodents obtained from reliable sources generally require only a physical
examination on arrival. This need not, on a routine basis, necessarily involve the
veterinarian directly; however, s/he should be advised of any health or other problem
involving the well-being of the shipment noted by either the technician or researchers
involved in receipt of the animals.

Information on method of transportation, the supplier's quality control profile, and the
animal's former environment should be available and, if not provided, should be
requested of each supplier.

Dogs, cats, non-human primates, and large domestic animals obtained from random
sources will usually be accompanied by only very limited information on their genetic
and medical history. Examination and quarantine procedures for these animals must be
stringent; specific diagnostic testing and immunization procedures should be established
and carried out following institutional SOPs for each specific species.

Federal government quarantine regulations, where applicable, will vary with the species,
the source, and the condition of the animal. These regulations may be obtained from the
nearest district veterinary officer, Food Production and Inspection Branch, Agriculture
Canada.

3. Animal Care Personnel

We are morally responsible for any living thing that we cause to be dependent upon us,
including animals used in research, teaching and testing. Exemplary standards of humane

37
care and treatment must be exercised by each person associated with captive animals.
Satisfactory buildings and equipment are, of course, necessary; however, of even greater
importance is the common sense and concern of all levels of personnel involved in the
care and use of experimental animals. Only the general responsibilities of such persons
can be outlined here; detail will vary with the institution and programs involved.

In every institution utilizing animals, a competent professional staff member must be

clearly designated with overall responsibility for care of experimental animals.

a) Chief or Director

The Chief or Director of animal care in large institutions is responsible for administration
of the animal care facility and should be directly responsible to a senior administrative
official; and should also be an ex-officio member of the ACC. This individual should be
qualified in an appropriate scientific discipline, possess considerable experience with a
variety of species, understand the requirements of research and be a competent
administrator.

This individual should be responsible for establishing or promoting participation in

educational programs which will, at the technical level, improve the quality and
efficiency of animal care and, at the professional level, assist in the proper training of
prospective investigators in the use of laboratory animals.

The responsibility to ensure that animals used for research, teaching and testing are of a
high quality, appropriate to the requirements of the investigator or teacher should also rest
with this individual.

In small institutions, the duties of "Director of Animal Care" may be assigned on a part-
time basis.

b) The Scientist-Teacher

The scientist-teacher should have knowledge of the characteristics, care and handling of
the species being utilized, and be committed to comply with the guidelines for care and
ethical use of animals as contained in this Guide.

The primary responsibility for the prevention of pain and discomfort in the experimental
situation lies with the investigator.

c) Animal Care Staff

It is the responsibility of the institution, through its ACC, to assure that its technicians
have the opportunity to become as well-qualified as possible. As in any field, opportunity
for continuing education should be provided, and all personnel should be encouraged to
participate.

38
Support staff are in a prime position to ensure both high quality animal care and the
success of an experiment through their diligence and daily observation of their charges. It
is noteworthy that distress in animals is not limited to the experimental situation, but may
result from improper housing and handling. Animals respond in a positive manner to
gentleness and considerate attention from their attendants.

Staff working with experimental animals may be involved either in their daily
maintenance or in the performance of primary experimental procedures or a combination
of these roles.

It is important that these persons be skilled and conscientious, as the well-being of the
animal and the success of the experiment on a day-to-day basis, are in their hands. In
addition to in-house training, formal courses in animal health and animal care technology
are now offered at 15 community colleges across Canada. Organizations exist which
provide training programs for the animal technician, leading to certification in his/her
field; e.g., Canadian Association for Laboratory Animal Science (CALAS). A co-
operative venture between employers and a community college has recently been
undertaken (Benn and McLaughlin, 1992).

C. REFERENCES

AGRICULTURE CANADA. Publication 1771/E. Recommended code of practice for the

care and handling of pigs. Communications Branch, Agriculture Canada, Ottawa, Ont.
K1A 0C7. 1984.

IBID. Publication 1898/E. Recommended code of practice for the care and handling of
farm animals--pigs. In Press.

IBID. Publication 1819/E. Recommended code of practice for the care and handling of
mink. 1988.

IBID. Publication 1821/E. Recommended code of practice for the care and handling of
special fed veal calves. 1988.

IBID. Publication 1757/E. Recommended code of practice for the care and handling of
poultry from hatchery to processing plant. 1989.

IBID. Publication 1831/E. Recommended code of practice for the care and handling of
ranched fox. 1989.

IBID. Publication 1853/E. Recommended code of practice for the care and handling of
dairy cattle. 1990.

IBID. Publication 1870/E. Recommended code of practice for the care and handling of
farm animals--beef cattle. 1992.

39
ALDHOUS, P. Lax enforcement of animal rules alleged. Nature 1990; 345: 190.

ANIMAL WELFARE ACT. 7 U.S.C., 2131-2157. Rev. 1985.

ANON. Animal care council outlined. Toronto Globe and Mail August 1, 1967.

ANON. Pound law to the boneyard. Toronto Star 1987 January 23.

ANON. (editorial) A Criminal Code fit for our time. Toronto Star May 21, 1988a: D2.

ANON. It's time to reform the Criminal Code. Ottawa Citizen 1988b May 21: B2.

ANON. Take Zundel's 'false news' crime off the books, law commission urges. The
Montreal Gazette May 20, 1988c.

ANON. CFHS President announces campaign for lab animal legislation. CFHS
(Canadian Federation of Humane Societies) Animal Welfare in Focus 1989 Fall: 1.

ANON. Special investigation--Professor Wilhelm Feldberg, CBE, FRS. Advocates for

Animals Quarterly, 79th Annual Report, year ending Dec. 31, 1990: 21-32.

ANON. Quebec considers research animal law. CCAC (Canadian Council on Animal
Care) Resource 1991a; 15(2): 3.

ANON. MRC reports on the Feldberg case. Vet. Rec. 1991b; 128(6): 120.

ANON. More monkey business. Nature 1991c; 349: January 3: 5.

ANON. The day the mice roared. Animal Activist Alert 1992a; 10(1): 1.

ANON. Legislation introduced in 102nd Congress. Washington Vet. News 1992b; 16(1):
4-5.

ANON. Animal Enterprise Protection Act is law. NABR (National Association for
Biomedical Research) Update 1992c; 13(16): 1.

ANON. Decline in animal experiments halted. Vet. Rec. 1992d; 131(5): 86.

BALLS, M. Animals (Scientific Procedures) Act 1986: The Animal Procedures

Committee. ATLA (Alternatives To Live Animals) 1986; 14: 6-13.

BALLS, M. (editorial) Animal experimentation: the weighing of benefit and suffering.

ATLA (Alternatives To Live Animals) 1989; 16: 212.

BALLS, M. (editorial) Has the Animals (Scientific Procedures) Act 1986 failed? ATLA

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(Alternatives To Live Animals) 1990; 17(4): 284.

BENN, D.M. and McLAUGHLIN, S.M. Training programs for personnel working with
laboratory animals. In: Animal care committees: Role and responsibilities. Ottawa, Ont.:
Canadian Council on Animal Care, 1992: 43-47.

BROWN, L. Cruelty to animals--the moral debt. Basingstoke: McMillan Publications,

Ltd., 1988.

CANADIAN PRESS. Quebec companies supply stray pets for research in U.S. Toronto
Globe and Mail 1985 July 31.

CHURCHWARD, P. The vivisectors' guide to pain. Liberator June/July 1986: 11.

COMEAU, P. Bill would keep pets from labs. Toronto Sun 1987 January 28.

DUQUETTE, M. Quebec's pound animals. Canadian SPCA Animag 1986 Fall: 10-11.

FISHER, C. Stepping up the pace. Liberator Autumn 1990: 20-21.

GILMAN, J.P.W. Safety testing of toxic substances. A survey. Ottawa, Ont.: Canadian
Federation of Humane Societies, 1980: x.

HAMPSON, J. The secret world of animal experiments. New Scientist April 11, 1992:
24-27.

HARVEY, I. Researchers, activists in battle over animals. Sunday Sun March 25, 1990:
36.

HEALTH RESEARCH EXTENSION ACT OF 1985, Public Law 99-158, November 20,
1985, "Animals in Research."

HEFLIN, H. S.544. Animal Welfare Information Center Newsl. 1992; 3(3): 1, 7-8.

HENSHALL, P., SCOTT, J.M. and SCOTT, J.L., eds. The use and care of animals in the
classroom. Cooksville, Canada: Peel Board of Education, 1986.

HOLLANDS, C. The Animals (Scientific Procedures) Act 1986. Lancet July 5, 1986: 32-
33.

INSTITUTE OF LABORATORY ANIMAL RESOURCES. Principles and guidelines for

the use of animals in precollege education. ILAR News 1989; 31(3): A2-A3.

LAW REFORM COMMISSION OF CANADA. Report 31. Recodifying criminal law.

Ottawa, Ont.: Law Reform Commission, 1987: 97-99.

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LOVE, J. Primate diseases. In: Johnston, C.E., ed. Proc. Animal diseases workshop.
Halifax, November 15, 1980. Halifax: Atlantic Provinces Inter-University Committee on
the Sciences, 1980: 87-94.

McANDREW, B. Humane society spent $200,000 fighting law. Toronto Star 1987 May
21.

McGIFFEN, H. and BROWNLEY, N., eds. Animals in education. The use of animals in
highschool biology classes and science fairs. Washington, DC: Institute for the Study of
Animal Problems, 1980.

McKIE, D. The Animals (Scientific Procedures) Act 1986. Lancet March 1, 1986: 513.

MENCH, J.A., MAYER, S.J. and KRULISCH, L. The well-being of agricultural animals
in biomedical research. Bethesda, MD: SCAW (Scientists Center for Animal Welfare),
1992.

MERVIS, J. U.S. judge throws out laboratory rules for dogs, primates. Nature 1993,
362(6415): 5.

MYERS, C. U.S. eases proposed regulations on care of laboratory animals; Researchers

are relieved, but welfare groups are critical. Chronicle of Higher Education 1990
September 5: A20.

MYERS, C. Universities must draw up written plans for their treatment of research
animals. Chronicle of Higher Education 1991 April 17: A21, A24.

NATIONAL ASSOCIATION FOR BIOMEDICAL RESEARCH. State laws concerning

the use of animals in research. Washington, DC: NABR, 1991.

NATIONAL ASSOCIATION OF BIOLOGY TEACHERS. The responsible use of

animals in biology classrooms. Including alternatives to dissection. NABT, 19-11250
Roger Bacon Drive, Reston, VA, 22090; 1990.

OLFERT, E.D., FINLEY, G.G., LANIEL, A.-M., LONGAIR, J.A. and ROWSELL, H.C.
The proposed new cruelty to animals law-Implications for veterinarians. Can. Vet. J.
1989; 30: 115-116.

ORLANS, F.B. Animal care from protozoa to small mammals. Menlo Park, CA; Don
Mills, Ont.: Addison-Wesley Publishing Co., 1977.

ORLANS, F.B., SIMMONDS, R.C. and DODDS, W.J., eds. Effective animal care and
use committees. Bethesda, MD: SCAW (Scientists Center for Animal Welfare), 1987.

OWEN, D.G. Parasites of laboratory animals. Laboratory handbooks No. 12. (Published
for Laboratory Animals Ltd.) London: Royal Society of Medicine Services Ltd., 1992.

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REINHARDT, V. Transport-cage training of caged rhesus macaques. Anim. Tech. 1992;
43(1): 57-61.

RICHTER, C.B., LEHNER, N.D.M. and HENRICKSON, R.V. Primates. In: Fox, J.G.,
Cohen, B.J. and Loew, F.M., eds. Laboratory Animal Medicine. Orlando, San Diego, San
Francisco, New York, London, Toronto, Montreal, Sydney, Tokyo, Sao Paulo: Academic
Press, 1984: 298-321.

ROWSELL, H.C. Report of a feasibility survey on the establishment of a Canadian

Council on Animal Care. Ottawa, Ont.: Association of Universities and Colleges of
Canada, 1967.

ROYAL SOCIETY FOR THE PREVENTION OF CRUELTY TO ANIMALS. Animals

in schools. 2nd Ed. West Sussex: RSPCA, 1985.

RUSSELL, W.M.S. and BURCH, R.L. The principles of humane experimental technique.
Springfield: Charles C. Thomas, 1959.

SANDERSON, T.A.H. (letters) CVMA issues statement on use of pound animals in

medical research. Can. Vet. J. 1986; 27(5): A7-A8.

SCHWINDAMAN, D.F. Regulatory requirements for exercise of dogs. In: Mench, J.A.
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SECORD, D.C. The Alberta research animal legislation. Can. Vet. J. 1974; 15(3); 92-94.

SHEPPARD, R. Little hope to save pound pups. Toronto Globe and Mail 1987 January
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SMITH, M. (editorial) The weighing of benefit and suffering. FRAME (Fund for the
Replacement of Animals in Medical Experiments) News 1988; 20: 1-2.

SMYTHE, D.H. Alternatives to animal experiments. London: Scolar Press-Royal

Defence Society, 1978.

SPECIAL COMMITTEE ON THE CARE OF EXPERIMENTAL ANIMALS. Report.

Ottawa, Ont.: National Research Council of Canada, 1966.

SPIRA, H. Here's what you can do to make a difference. Factory farming. Animal Rights
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STEWART, W.A. Debates in the Province of Ontario Legislature, June 17, 1969b: 5722.

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WILSON, D. Researchers irked by move on strays. Toronto Globe and Mail July 9, 1986.

Chapter II - Laboratory Animal Facilities

II. LABORATORY ANIMAL FACILITIES

A. INTRODUCTION

A laboratory animal facility must facilitate research by minimizing undesirable

experimental variables while providing for the physiological, social and behavioural
requirements of the animal. Different research projects and/or different species of animals
often require differing facilities and environments. To accommodate such needs, an
animal facility must have separate areas for carrying out different functions, specialized
rooms and equipment, and closely controlled environments.

Animal facilities providing the appropriate environment are expensive to build. It is,
therefore, imperative that every effort be made to ensure that any proposed new facility is
programmed, designed, and built to meet the size and scope of current animal use, and
yet to be versatile enough to allow flexibility in the years to come.

A number of alternative design approaches to achieve any given functional need are
available. For example, the Handbook of Facilities Planning, Volume 2: Laboratory
Animal Facilities (Ruys, 1991), is a useful reference for the planning phase. Other
references and assistance may be obtained from the Canadian Council on Animal Care
(CCAC). It is strongly recommended that the CCAC be involved at an early stage in the
planning phase and that plans be evaluated by the Council before the start of
construction.

B. LOCATION

Animal facilities should be located so as to minimize public access or through-traffic, as

well as the movement of animals, cages, waste, etc., through public corridors and
elevators. The facilities should be readily accessible by animal users yet easily secured.
Direct access to the outside for deliveries and disposal is desirable. Facilities located on
higher floors should be accessed by a minimum of two elevators, one for clean and one
for dirty materials, unless appropriate measures are taken to clean and sanitize a single
elevator following the transport of dirty materials. For very small and/or satellite

44
facilities, alternative precautions to minimize contamination may be acceptable.

C. MECHANICAL SERVICES

Heating, ventilation and air conditioning systems for animal facilities are usually quite
sophisticated and costly (see also The Environment). Placement of these systems should
allow servicing to proceed with a minimum of disturbance to the animals and the work
patterns in the facilities. This may be accomplished by placing mechanical services on a
separate service floor immediately above the animal facilities so that maintenance does
not require entry into the animal facilities. It is, however, more common to locate the
mechanical systems in the ceiling space between floors. In this location all access to the
mechanical systems should be from the corridors, and not from the animal rooms or
restricted zones such as biohazard areas.

D. DESIGN

The size of animal rooms should be based on the species to be maintained, and a multiple
of the size of pens, cages or cage racks to be contained, allowing for adequate ventilation
and servicing. Animal rooms should be designed for ease of sanitation and hence should
have a minimum of built-in equipment. In many cases, a single, small sink for hand
washing may be all that is required. The placement of the animal rooms and ancillary
rooms will depend on the species, experimental use and microbial quality. The design
should facilitate traffic flow from cleaner to dirtier areas. Rooms requiring frequent
access by investigators should be located near the entry to the facilities to minimize
traffic.

E. MAJOR FUNCTIONAL DIVISIONS

The design of an experimental animal facility should take into consideration the needs of
the experimental animal and the requirements and convenience of the scientists and
technical staff. Good animal care facilities must provide for several separate functions
and sometimes highly specialized areas (Clough, 1986; Home Office, 1986). Animal
holding rooms should be separate from experimental rooms. Important aspects of good
design are provision for efficient and effective sanitation, efficient work traffic patterns
and orderly expansion. The following identifies the major functional areas in an ideal
animal facility.

1. Animal Reception Area

The reception area should be situated so that animals entering it do not pass through
holding or experimental areas. Similarly, waste material should not pass through the
receiving area. It should provide sufficient space for the uncrating and initial examination
of animals as well as for holding them under appropriate environmental conditions until

45
they are relocated either in the conditioning area or one of the animal rooms.

2. Conditioning Rooms
Conditioning rooms are ones in which animals may be maintained for detailed health
examination, observation and conditioning in preparation for experimentation. The
availability of a proper conditioning room(s) is particularly important where random-
source animals are being acquired (e.g., some dogs, cats, non-human primates, and
animals from the wild). Under certain circumstances, where space permits, it may be
possible and even desirable to immediately house animals in an experimental room,
provided the animals are derived from a single source and contact with other animals can
be avoided.

3. Holding Rooms

Separate animal holding rooms should be available for each species, from each source,
and for each investigator's project. Consequently, it is usually better to have many small
rooms rather than a few large ones. Exceptions can be made where investigators are using
the same species from the same source for different projects (e.g., antibody production in
rabbits). Mixing should be limited to compatible social and health status groups within a
single species. Where mixing of species is necessary, some degree of isolation may be
achieved by specialized room design, equipment and/or cage selection. Cross-
contamination can be minimized when controlled airflow cubicles, portable laminar
airflow units, and various forms of isolator cages are used. The use of radioisotopes,
infectious agents and highly toxic substances requires special holding rooms. Rooms
suitable for special purposes may also be required (e.g., breeding colonies, controlled
environmental studies and for the housing of both farm and wild animals).

It is important, when designing holding rooms to consider possible future uses of these
facilities. Where animal use has been consistent over the years, it may be acceptable to
design all animal rooms for specific species use. However, in many facilities, animal use
fluctuates considerably, making flexibility extremely important. A flexible holding room
is one which meets the acceptable requirements for housing different species.

4. Quarantine/Isolation Rooms

Within the facility, but apart from the conditioning area, quarantine/isolation rooms may
be required to separate sick animals or those animals returned to the facility after use in
an investigator's laboratory.

5. Experimental and Treatment Facilities

Experimental manipulations should not be carried out in animal holding rooms, unless
mandated by experimental design or containment needs and approved by the Animal Care
Committee (ACC). Separate facilities should be available to allow for surgery,
euthanasia, etc.; however, not all need to be located within the animal facilities. The
animal holding rooms should therefore be located as conveniently as possible to the

46
research and teaching laboratories.

Animal facilities may include rooms for some or all of the following: pre-surgical
preparation, surgery, post-operative recovery (see also Standards for Experimental
Animal Surgery), radiology, necropsy, diagnostic services, special diet preparation,
dispensary, etc. The design and organization of special facilities will depend on their
intended scope and use; however, even very modest facilities will usually need to provide
a special area or procedures room for minor surgery and/or treatments, and a separate
necropsy room.

Separate diagnostic areas for laboratory animal diseases may not be feasible for smaller
institutions. In such cases, arrangements for provision of such services should be made.

6. Support Facilities

a) Facilities for Washing and Sterilizing Equipment

Facilities for washing and sterilizing equipment and supplies should be designed for this
purpose and be located so as to minimize disturbance to animals, staff, and neighbours.
Ventilation should be sufficient to prevent odours, excess heat and steam from affecting
the rest of the facility. Sinks for hand-washing and for cleaning specialized pieces of
equipment are useful. Large, deep sinks are useful. Autoclaves and other special
equipment may be located in this area. Ideally, the wash-up area should provide for the
separation of clean and dirty equipment. If spray washing of either cages or racks is to be
used, provision of a walled-off bay with hot and cold water and disinfectant dispenser is
recommended.

b) Waste Disposal

The waste disposal area should provide for proper storage of animal material, excrement,
soiled bedding, etc. Waste awaiting collection should be placed in a dedicated refrigerated
container or cold room. Waste stored outside the facility should be in secure covered
containers. Facilities must comply with local bylaws governing waste storage and
disposal. Toxic, infectious or radioactive waste handling must comply with institutional,
federal (HWC/MRC, 1990) and other regulations (see also Occupational Health and
Safety).

c) Food and Bedding Storage

Small quantities of food and bedding may be stored in an animal room in suitable,
covered containers. Separate cool (<15C), dry, vermin-proof facilities should be available
for the storage of food to minimize spoilage and contamination. Food for farm animals,
such as hay, may contain vermin and should be isolated from the food and bedding of
other laboratory animals.

d) Equipment Storage

47
Lack of sufficient storage space is one of the most frequent and more serious deficiencies
encountered in facility design. Equipment storage should not be permitted in halls,
corridors, or in rooms housing animals. Even clean equipment designated for use in an
animal room should not be brought into the room until required. Areas used for storing
clean equipment should be separate from those where soiled equipment is received. For
the average facility, 11% storage space (of net space) has been estimated as adequate.
This proportion will need to be increased up to 20% or more in facilities handling
multiple species under differing barrier conditions.

7. Personnel, Office and Reception Areas

These functional areas may or may not be combined. It is preferable to have these
adjoining, but not within the animal facilities. Enough office space is required to
accommodate all administrative staff, occasionally technicians, and the extensive records
that must be maintained.

8. Facilities for Personnel

Personnel facilities should encourage high standards of personal hygiene by providing

staff with easily accessible changing rooms with lockers, showers, sinks and toilets.
Depending on the design of the facility, these may have to be replicated in different
zones. Suitable protective clothing should be supplied (see also Occupational Health and
Safety).

Facilities should be provided for staff rest periods, lunch, and for meetings. It is
preferable that these areas are adjoining, but not within the animal holding areas. An
information centre for staff (which could include books, journals, newsletters, catalogues,
and other related materials) would be helpful.

F. SECURITY

Access to experimental animal facilities must be restricted in order to assure consistent

environmental control and to minimize interferences which might alter experimental
results. Entry and exit should be limited and the facilities maintained secure at all times.
Access should be allowed only to those who have a recognized need to enter. Where a
large number of investigators are using the same facilities, it is often advisable to have
individual room locks. Electronic access control systems are available.

G. CONSTRUCTION GUIDELINES FOR ANIMAL ROOMS

1. Floors and Drains

Floors should be seamless, durable, non-slippery, impervious to water and easy to

48
disinfect. They should be coved to the walls to eliminate any sharp corners. They should
slope towards any floor drain(s) and the proper level of this slope should be verified in all
new construction. Recommended minimum pitch of sloped floors is 2.1 cm/m (0.25"/ft).
Special attention should be given to ensuring that this critical component of floor
construction is properly carried out.

It is recommended that drains be provided with a flush mechanism so that a clean water
seal (i.e., clean water in the trap) can be maintained. However, care should be taken to
assure that the location of the flush mechanism does not interfere with cage or pen
placement. Drains should be provided with an adequate cover and pitch basket trap.
Drain and sewage lines should be at least 10.5 cm (4") in diameter. Where dog excreta is
washed down the drain, the diameter should be 15.0 cm (6"). Floor drains used for waste
disposal should be placed at the end of the main drainage line. Drains should be checked
regularly to ensure proper functioning, an effective water seal, and absence of insects.
When not in use, they should be capped and sealed.

Floor drains may not be required in rooms designed solely to house small species. Instead
wet vacuum devices which permit sweeping and mopping with disinfectants or a cleaning
compound may be used.

2. Walls and Ceilings

Walls should be of an impervious material, free of cracks, damage resistant, and easily
cleaned and disinfected. With these kinds of surfaces, noise reduction is difficult. Walls
need not be as resistant as the floor, provided that they are protected by a cove or bumper
guard. Pipe and service sleeves should be adequately closed off and sealed so as to
exclude vermin.

Ceilings within all rooms should be seamless and free of cracks, with ceiling-wall joints
well sealed. In some corridors, it may be necessary to use ceiling tiles in order to allow
access to the mechanical systems. These tiles should be of a type which can be easily
sanitized and which prevent the entry of vermin into the ceiling space.

3. Doors

Animal room doors should be designed and built to exclude vermin. Self-closing, metal
or metal-covered doors with obscurable viewing windows and kick plates are preferred. A
replaceable sweeper pad should be installed along the bottom if clearance exceeds 0.32
cm (1/8"). Recommended minimum door sizes are 107 cm (42") wide and 213 cm (84")
high to allow free passage of equipment.

4. Windows

Exterior windows interfere with temperature control due to radiation and conduction
which may jeopardize animal health and research results. They also interfere with
photoperiod control. If windows are already present these should be designed or altered

49
to minimize the above effects and to maximize cleanliness.

5. Corridors

Corridors should be strategically located to facilitate traffic flow for the desired work
patterns. It may be more efficient to divide the animal facilities into zones with single
corridors than to use a double corridor (clean/dirty; supply/return) system.

Design standards for corridor floors, walls, drains, coving, bumper guards, etc., should
duplicate those for animal rooms. Traffic corridors should be at least 1.82 m (6') wide.
Other corridors should be wide enough to allow free movement of personnel and
equipment. Walls should be free of any projecting fixtures up to a height of 213 cm (84");
alternatively, these fixtures should be adequately protected with guards. Exposed corners
should be protected with steel plates or similar durable material. All guards and fixtures
should be sealed to exclude vermin. Corridors leading to noisy areas should have double
doors or similar noise-trapping devices.

6. Services

Service lines should be located on the floor above the animal facilities or in the ceiling
space above the corridors so as to eliminate in-room maintenance. Separate hot and cold
water lines should be supplied to each animal room for hand washing, cleaning, and
automatic watering. Every animal room should have at least one electrical outlet; these
should be water, insect and explosion-proof. Switches and thermostats should be
similarly designed. An emergency power source should be available in case of a power
failure.

H. CAGING

The size of caging chosen to house each species should be appropriate for that species
(see Appendix I).

Cages and pens must not only confine the animals securely, but also ensure their comfort
and safety by permitting normal postural and behavioural adjustments, and provide for
environmental enrichment. Animals which are social by nature should not be singly
housed unless this is a necessary requirement of the research protocol, and approved by
the ACC (see also Social and Behavioural Requirements of Experimental Animals).

Cages must provide for adequate ventilation, satisfactory viewing and easy access to the
animal. Food and water delivery systems should be designed and located so as to allow
the animal ready access, but prevent contamination with excrement. Cage design should
facilitate cleaning and disinfection.

The intensity of light perceived by the animal, the level of noise to which it is exposed,
the ventilation and temperature of its microenvironment are affected by cage design and

50
material. Considerable care should be used when choosing the appropriate caging for a
particular species and usage. Caging for animals other than the conventional laboratory
species requires special consideration.

Unless contra-indicated by the nature of the research (e.g., nutritional studies) solid
bottom cages should be chosen (over suspended wire caging) for rodents and guinea pigs
in that they permit creation of microenvironments and facilitate provision of
environmental enrichment (see also Social and Behavioural Requirements of
Experimental Animals).

1. Shoebox Cages

The shoebox cages used mainly for small rodents are particularly suited for breeding
purposes. They are usually made of plastics such as polycarbonate, polystyrene, and
polypropylene. Polycarbonate is clear, autoclavable, and resistant to most disinfectants.
Polystyrene and polypropylene do not withstand high temperatures well. Polypropylene
cages are translucent and offer animals more privacy, which may be beneficial for some
breeds or wild species. However, opaque cages should not be placed on shelves above
eye level since the animals within cannot be readily observed.

A contact bedding (e.g., woodchip, ground corncob, etc.) is used in the bottom of
shoebox cages, allowing an animal to form its own microenvironment. These cages are
considered comfortable for the animal, and the cage of choice for breeding. However,
animals in these cages are in contact with their own excreta and airflow is restricted.
Therefore, it is important to clean the cages frequently. Filter caps restrict the airflow
even more if cages are not individually ventilated. Faster buildup of ammonia, carbon
dioxide and moisture necessitates more frequent cleaning (up to three times per week
may be required). Shoebox cages can be fitted with wire grid floors for certain projects
which require that there be no contact with excreta.

2. Larger Solid Bottom Caging

Large plastic tubs have been used quite successfully for group housing guinea pigs and
rabbits. These tubs must be strong enough to support the weight of the animals contained,
have rounded corners to facilitate cleaning and be resistant to disinfectants. These are
used with a contact bedding.

3. Suspended Cages

Suspended cages may be top or front opening. Most top opening suspended cages use the
rack shelves as the top for the cage. The top opening cages are used primarily for smaller
rodents, whereas the front opening cages are better suited to guinea pigs, cats, dogs,
rabbits and non-human primates (NHP).

Most suspended cages have a floor of wire mesh, steel rod, perforated metal or plastic,
above a collection tray or solid floor. It is extremely important that the size of the floor

51
perforations be appropriate for the species housed. They should be large enough to permit
excreta to freely pass through, but small enough to prevent foot and leg injuries. The
gauge of wire should support the animal's weight without sagging. Floors should be
designed so the animal's feet can grip during movement, so as to minimize slipping. Wire
mesh floors are not suitable for guinea pigs nor for use in rodent littering cages.

In suspended cages, animals are not in contact with their own excreta and the cages are
usually well-ventilated. The pans or trays can be cleaned more frequently than the cage,
resulting in less disturbance of animals. The animals, however, do not have the
opportunity to form their own microenvironment, and so control of the room environment
becomes more critical.

It is recommended that these cages be fabricated from stainless steel or other woven
metal alloys, corrosive resistant plastic and/or in the case of some front opening cages,
fibreglass. Fibreglass is strong, warm-feeling, and more sound-resistant than other
materials, making it especially suitable for post-operative care. NHP and cats should be
supplied with one or more resting boards or perches at different levels. A squeeze device
built into the cage facilitates restraint of NHP.

4. Other Cages

Many cages are designed to meet specific requirements. Examples include metabolism
cages, mechanical exercise cages, gang cages, transfer cages, restraint cages and walk-in
cages (used for housing groups of animals).

Additional information on housing large domestic animals and fowl may be found in
Agriculture Canada's Canadian Farm Building Handbook (Agriculture Canada, 1988)
(see also Farm Animal Facilities and Environment) as well as Social and Behavioural
Requirements of Experimental Animals in this volume. Guidelines for the use of
agricultural animals have also been published in the U.S. (Curtis, 1988).

For information on cages for wild animals, contact the Secretary-Treasurer of the
Canadian Association of Zoological Parks and Aquariums, c/o Metro Toronto Zoological
Society, P.O. Box 280, West Hill, Ontario M1E 4R5.

Information on shipping crates and transport cages for a wide range of domestic, wild and
laboratory animals may be obtained from the most recent volume of Live Animals
Regulations (1992) of the International Air Transport Association (IATA), 2000 Peel
Street, Montreal, Quebec H3A 2R4.

All cage types must take into consideration the well-being of the animal(s) during its
confinement.

I. REFERENCES

52
AGRICULTURE CANADA. Publication 1822/E. Canadian farm building handbook.
Communications Branch, Agriculture Canada, Ottawa, Ont., K1A 0C7. 1988.

CLOUGH, G. The animal house: Design, equipment and environmental control. In:
Poole, T.B., ed. UFAW (Universities Federation for Animal Welfare) handbook on the
care and management of laboratory animals. 6th Ed. New York, NY: Churchill
Livingstone Inc. 1986: 108-158.

CURTIS, S.E., ed. Guide for the care and use of agricultural animals in agricultural
research and teaching, 1988. Consortium for Developing a Guide for the Care and Use of
Agricultural Animals in Agricultural Research and Teaching, 309 West Clark Street,
Champaign, IL 61820.

HEALTH AND WELFARE CANADA/MEDICAL RESEARCH COUNCIL. Laboratory

biosafety guidelines. Ottawa, Ont.: Office of Biosafety, Laboratory Centre for Disease
Control, Health and Welfare Canada, 1990.

HOME OFFICE. Code of practice for the housing and care of animals used in scientific
procedures. Act Eliz. II 1986 c.14 Section 21, Animals (Scientific Procedures) Act.
London: Her Majesty's Stationary Office 1986: 4-8.

INTERNATIONAL AIR TRANSPORT ASSOCIATION. Live Animals Regulations. 19th

Ed. Montreal, Quebec: IATA, 1992.

RUYS, T., ed. Handbook of facilities planning. Laboratory animal facilities. New York,
NY: Van Nostrand Reinhold, 1991: 2.

Chapter III - The Environment

III. THE ENVIRONMENT

There are many physical, chemical, and biological factors which may influence
experimental animals and thus modify the results of the investigations (Melby, 1983;
Small, 1983). The experimental results obtained are, in principle, only valid for the
conditions under which they were obtained and only useful for comparison if all the
relevant information concerning experimental conditions is made available.

Among the environmental factors which should be recorded for possible inclusion in
scientific reports are: temperature (C and range), relative humidity (% and range) and
whether or not these are regulated; air exchanges/hour, proportion of fresh and
recirculated air, and gas or particle concentrations in the air; lighting (natural and/or
artificial, photoperiod, and intensity); water type, quality, and pretreatment; bedding type,
quality, and pretreatment; housing density; housing equipment; and physical measures to
protect microbiological status. The microbiological status of the animal should be
reported [conventional, Specific Pathogen Free (SPF) for stated pathogens, or gnotobiotic

53
with microorganisms specified] (WCBCLA, 1985).

A. CLIMATE CONTROL

Environmental requirements vary with the species and the experimental protocol.
Environmental parameters are usually measured at the level of the room. More important,
however, is the microenvironment established at the cage level, since the conditions
between the two may differ dramatically (Woods, 1980; Corning and Lipman, 1992). A
summary of some environmental parameters for individual species is given in Appendix
I.

The design of the animal facility should permit adjustment of environmental controls to
meet the needs of the species and the experimental protocol. Ideally, each animal room
would be controlled independently. In facilities not originally constructed with this
capability, this ideal could be approached through proper management and the installation
of ancillary automatic light timers, rheostats, thermostatically controlled exhaust fans,
humidifiers, and air conditioning units.

1. Temperature

Published data on optimal temperatures for housing laboratory animals are variable
(CCAC, 1984; Clough, 1984; NRC, 1985). All these were considered when the guidelines
of the Council of Europe were formulated (European Convention, 1986).

It is essential that emergency equipment be available to maintain environmental

temperatures, particularly in rooms housing small laboratory animals, fish, and non-
human primates (NHP).

In special cases, for example, when housing very young or hairless animals, higher room
temperatures than those indicated may in Appendix I be required.

Animal room temperatures should be monitored daily, preferably by continuous

recording. A less-costly alternative is the use of a maximum/minimum thermometer
which is examined and re-set daily; however, this does not indicate how long the room
was held at a particular temperature, knowledge of which is extremely important
(McSheehy, 1983). If the experimental protocol or management practices require that an
animal be housed at temperatures outside the recommended range, an adequate time
should be given for adaptation to occur (Baker, Lindsey and Weisbroth, 1979). The
temperature of the microenvironment should also be monitored. Factors affecting
temperature in the cage include the type of cage and bedding or nesting material, the use
of filter covers, age, sex, strain, species, and housing density (Woods, 1980; Corning,
1992).

Environmental temperatures and variability can affect animal research and testing,
influencing an animal's response to drugs, susceptibility to infectious disease, fertility,

54
production, feed and water intake, growth curves, and hematologic parameters (Baker,
Lindsey and Weisbroth, 1979; Lindsey, 1978; Yamauchi, 1981). Occasionally, optimal
temperature for the research animal is not the most comfortable for personnel; however,
human preferences should not compromise the experimental requirements or the health
and comfort of the animal.

2. Humidity

Most laboratory animals prefer a relative humidity around 50%, but can tolerate a range
of 40-70% as long as it remains relatively constant and the temperature range is
appropriate (Clough, 1987). Discomfort results when humidity levels adversely affect the
animal's ability to maintain thermal homeostasis. In facilities where humidity is difficult
to control within an acceptable range, dehumidification or humidification devices may
need to be installed.

Humidity levels can affect experimental results by influencing temperature regulation,

animal performance, and disease susceptibility.

3. Ventilation

Ventilation influences temperature, humidity, and gaseous and particulate contaminants in

the animal cage and holding room. The design of the building ventilation system should
permit the maintenance of these parameters within acceptable limits. The actual
ventilation rate required varies with age, sex, species, stocking density, frequency of
cleaning, quality of incoming air, ambient temperature and humidity, and construction of
primary and secondary enclosures, among other factors. Draft-free air exchanges in the
range of 15-20 per hour are commonly recommended for rooms containing small
laboratory animals under conventional housing conditions (Clough, 1984). Achieving
these rates does not guarantee adequate ventilation at the cage level, particularly if filter-
tops are used (Keller, White, Sneller et al. 1989). Laminar flow units and rooms provide
good ventilation with an unidirectional airflow having few eddy currents. These systems
may effectively isolate cages within them, controlling the spread of odours and airborne
pathogens (Phillips and Runkle, 1973; McGarrity and Coriell, 1976).

Differential pressures can be used to inhibit the passage of pathogenic material between
rooms. Higher pressures are used in clean areas relative to dirty or biohazardous ones, in
order to minimize contamination (Hessler and Moreland, 1984). In facilities where
containment or exclusion of airborne microorganisms depends in part on differentials in
air pressure, inclined manometers or magnehelic gauges can be used to measure the
difference between the high and low pressure areas in millimetres of water. Generally,
2.5-5.0 mm (0.1-0.2 in.) differential is maintained (Small, 1983).

The design of the ventilation system should take energy conservation into account
(Besch, 1980). Although total air exchange systems are preferable, they are not always
economical, especially in regions experiencing temperature extremes. Recirculating air
systems must be equipped with effective filters (and scrubbers, if necessary) to avoid the

55
spread of disease and to remove particulate and gaseous contaminants (e.g., NH3)
(Hessler, 1984).

4. Lighting

The three characteristics of light which can influence laboratory animals are intensity,
quality, and photoperiod. The lighting should provide good visibility and uniform, glare-
free illumination. Previous recommendations of 807-1345 lux (75-125 fc) at 76 cm from
the floor have been shown to cause retinal degeneration in albino rats (Belhorn, 1980;
NRC, 1985; Semple-Rowland and Dawson, 1987). The recommended level of 323 lux
(30 fc) approximately 1.0 m above the floor has proved sufficient for the performance of
routine animal care duties and does not cause rodent phototoxic retinopathy (Belhorn,
1980). A level of approximately 200 lux does not appear to cause retinal damage and has
been shown to be adequate for reproduction and normal social behaviour among most
rodents (Weihe, 1976). At this level, an additional light source on a separate switch is
needed to enhance illumination during caretaking activities.

The intensity experienced by animals housed close to the source may differ markedly
from that experienced by those farther away, because light intensity is inversely
proportional to the square of the distance from its source. Additionally, light intensity
within a cage is dependent upon cage type and construction, position of the cage on the
rack, and type of rack, and may vary markedly from front to back (McSheehy, 1983).
Light intensity can influence aggressiveness and the incidence of cannibalism in rodents
(Weihe, 1976; Fall, 1974). Gradual changes between dark and light periods allow time for
behavioural adjustment and the expression of crepuscular behaviours. Fish and
amphibians may take thirty minutes to make intra-ocular adjustments to changing light
intensities (Allen, 1980).

There are few studies on the effect of light quality, or spectrum, on laboratory animals. It
has been stated that animal room illumination should duplicate the characteristics of
sunlight as closely as possible. There is some disagreement over the necessity for this in
every case (Belhorn, 1980; Small, 1983). Among laboratory rodents, a light spectrum that
differs markedly from sunlight may reduce breeding efficiency, cause behavioural
abnormalities, and enhance spontaneous tumour development (Weihe, 1976). High levels
of ultraviolet (UV) light can induce cataracts in laboratory mice (Belhorn, 1980). The
wavelength to which guppies are exposed influences fecundity and affects development
and sex ratios in offspring (Mulder, 1971). Exposure to UV light may cause epithelial
damage in some species exposed to photosensitizing agents. Electromagnetic waves
outside the visible spectrum may influence behaviour and activity of laboratory rats
(Mulder, 1971). Light tubes which imitate the spectrum of sunlight are commercially
available.

Photoperiod is probably the most influential of light characteristics on laboratory animals.

Photoperiod influences the circadian rhythms seen in biochemical, physiological, and
behavioural aspects of the animal - patterns stimulated and synchronized through the
neuroendocrine pathway. The circadian cycle can affect the animal's response to drugs or

56
resistance to inoculated infectious organisms (McSheehy, 1983). The light/dark ratio can
affect reproductive performance and sexual maturity. It is suggested that, if a change
occurs in an animal's photoperiod, no experiments be conducted with that animal for at
least a week (Davis, 1978). If the light phase is interrupted by dark, there are few
significant effects; however, if the reverse occurs, endogenous rhythms can be
significantly skewed (Davis, 1978). This is one reason why automatic timers should
control light cycles in all animal rooms. Timer function should be monitored or hooked
into an alarm system. Additionally, any windows in an animal room should be
occludable.

Differences in light, temperature, and airflow between locations on a cage rack can affect
experimental results and should be minimized by either rotating cages through different
positions on a rack, or by assigning animals to cages based on a table of random numbers.

B. OTHER ENVIRONMENTAL FACTORS

1. Noise

The effects of noise on laboratory animals are related to its intensity, frequency, rapidity
of onset, duration and characteristics of the animal (species, strain, noise exposure
history). Species differ in their auditory sensitivity and susceptibility to noise-induced
hearing loss. Prolonged exposure to high levels of noise can cause auditory lesions in
animals. Although a maximum background noise of 85 db has been recommended
(Baker, 1979), adverse changes have occurred in rats exposed to intermittent noise at 83
db (Gerber, Anderson and Van Dyne, 1966). Exposure to uniform stimulus patterns may
lead more readily to hearing loss, whereas exposure to irregular patterns may be more
likely to cause disorders due to repeated activation of the neuroendocrine system
(Peterson, 1980).

Intense noise can cause alterations in gastrointestinal, immunological, reproductive,

nervous, and cardiovascular systems, as well as changes in development, hormone levels,
adrenal structure, blood cell counts, metabolism, organ weights, food intake, and
behaviour (Agnes, Sartorelli, Abdi et al. 1990; Bailey, Stephens and Delaney, 1986;
Fletcher, 1976; Kraicer, Beraud and Lywood, 1977; Nayfield and Besch, 1981; Pfaff,
1974; Gerber and Anderson, 1967). Sudden intense sound can elicit startle responses and
can precipitate epileptiform seizures in several species and strains of laboratory animals
(Iturrian, 1971; Pfaff, 1974). Ultrasound emissions can cause behavioural disturbances in
a variety of species (Algers, 1984). Although firm criteria for noise tolerance have not
been established for laboratory animals as for humans (Falk, 1973; Welch and Welch,
1970), unnecessary and excessive noise may be assumed to be an important experimental
variable and a possible health hazard.

Noise can be controlled in an animal facility through proper facility design and
construction, thoughtful selection of equipment, and good management practices.

57
Naturally noisy animals should be located where they minimally disturb quiet, noise-
sensitive species. Fire alarms which operate at low frequencies are audible to humans, but
do not disturb mice and rats. Telephones should not be placed in animal rooms. Many
noise sources in an animal facility emit ultrasound (Sales, Wilson, Spencer et al. 1988).
These include running taps and squeaking chairs. Efforts should be made to identify and
correct or shield these sources.

Noise can also disturb or harm animal care staff, researchers, and other nearby personnel.
It may be necessary to provide ear protectors in some areas such as dog, pig, or monkey
rooms, or the cage-washing facility.

2. Chemicals

Chemicals in the environment can adversely affect the laboratory animal in a variety of
ways. Inherently toxic compounds or toxic metabolites can have local and/or systemic
effects on virtually every system. Although most chemicals found in animal facilities
exert their major effect by altering hepatic microsomal enzyme activity, immune function,
or behaviour, allergens, mutagens, teratogens, and carcinogens have also been detected.
Their ultimate effect is modulated by the interplay between chemical factors
(concentration; physicochemical properties; duration, frequency, and route of exposure;
interaction with other agents) and host factors (species, age, sex, strain, nutritional status,
immune function, disease status) (Baker, Lindsey and Weisbroth, 1979).

Chemicals arrive in the microenvironment through air, water, food, bedding, and contact
surfaces. Common air pollutants include dust and bedding particles, ammonia,
disinfectants, pheromones, organic solvents, volatile anesthetics, insecticides, and
perfumes or deodorants.

The most common air contaminant in animal facilities is ammonia (NH3) resulting from
the decomposition of nitrogenous waste. Ammonia causes irritation of the respiratory
epithelium and increases susceptibility of rodents to respiratory mycoplasmosis
(Broderson, Lindsey and Crawford, 1976; Lindsey, Connor and Baker, 1978). Sub-
clinical pathological changes in the respiratory tract due to ammonia complicate
inhalation toxicity studies in laboratory rodents (Gamble, 1976). In humans, 25 ppm is
the level below which there are no harmful effects from an 8 hr/day, 5 day/week exposure
[American Conference of Government and Industrial Hygienists Threshold Limit Value
(TLV)]. The human odour detection threshold for ammonia is 8 ppm. In comparison, the
TLV is 17 mg/m3.

The animal's microenvironment must be checked as well as the room, because conditions
often differ significantly between the two (Corning and Lipman, 1992). Ammonia levels
build up when production components (species, sex, housing density, bedding) exceed
elimination components (cage design, air exchange, frequency of cleaning) (Serrano,
1971). Filter covers, which reduce air exchange at the cage level, can rapidly lead to
detrimental concentrations of NH3. Controlling NH3 within safe levels requires constant
attention to stocking density and to frequency of cage cleaning.

58
Perfume and deodorants should never be used to mask ammonia or other animal odours
in lieu of proper husbandry. These substances may be harmful to the animals (Baker,
Lindsey and Weisbroth, 1979; Pakes, Lu and Meunier, 1984). Volatile anesthetics should
be used only with proper scavenging equipment.

Chemicals can enter the animal's environment through the water. Other than checking for
bacterial contaminants, water quality is rarely monitored except for aquatic animals.
Chlorinated municipal water sources are commonly used. Over 700 organic compounds
have been isolated from such sources - 90% are natural decomposition products. These
may react with chlorine to produce chloroform (Pakes, Lu and Meunier, 1984). Inorganic
solutes, particularly copper (from copper pipe) and chlorine are especially hazardous to
aquatic organisms.

Food may be contaminated with heavy metals (e.g., lead, arsenic, cadmium, nickel,
mercury), naturally occurring toxins (e.g., mycotoxins, ergot alkaloids, pyrrolizidine
alkaloids, estrogenic compounds), agricultural chemicals (e.g., herbicides, pesticides,
fertilizers), and additives (e.g., antibiotics, colouring, preservatives, flavourings,
unintentionally incorporated drugs) (Baker, Lindsey and Weisbroth, 1979; Pakes, Lu and
Meunier, 1984; Silverman and Adams, 1983).

Chemicals found on contact surfaces include cleaning agents such as soaps, wetting
agents, detergents, solvents, and disinfectants (Burek and Schwetz, 1980). Unless
otherwise specified as safe according to the manufacturer's instructions, these substances
should be thoroughly rinsed from surfaces which will contact animals. The efficacy of the
rinse cycle of the cage-washer should be checked periodically.

Bedding materials, particularly wood products, may introduce naturally occurring volatile
oils, herbicides, pesticides, and preservatives into the animal's microenvironment. Other
possible contaminants include PCB's and antibiotics (Silverman and Adams, 1983).
Volatile hydrocarbons in cedar and pine shavings can induce hepatic microsomal
enzymes (Weisbroth, 1979).

3. Bedding

The choice of bedding materials and cage flooring profoundly affects the
microenvironment of small rodents. In most circumstances, contact bedding is
recommended. Most species should be provided with solid flooring and bedding prior to
parturition. Some desirable characteristics of contact bedding are listed below.

Bedding material should always be taken into consideration in designing an experiment

and should be uniform throughout the study because of its influence on behavioural and
physiological responses and on toxicity and carcinogenesis studies.

59
 DESIRABLE CRITERIA FOR RODENT CONTACT BEDDING (Kraft, 1980)
Moisture absorbent Unlikely to be chewed or mouthed

Dust free Non-toxic

Unable to support bacterial growth Non-malodorous

Inedible Nestable

Non-staining Disposable by incineration

Non-traumatic Readily available

Ammonia binding Relatively inexpensive

Sterilizable Fire resistant

Deleterious products not formed as a Remains chemically stable during use

result of sterilization
Manifests batch to batch uniformity
Easily stored
Optimizes normal animal behaviour
Non-desiccating to the animal
Non-deleterious to cage-washers
Uncontaminated
Non-injurious and non-hazardous to
Non-nutritious personnel

Non-palatable

Unsterilized materials are a possible source for the introduction of disease into rodent
colonies. Wild rodents enjoy nesting in packages of bedding, and cats will defecate in
loose bedding (Newman and Kowalski, 1973). Recommended bedding materials for each
species are discussed in Volume 2 of this Guide.

4. Population Density and Space Limitations

Population density and group size influence the physiological and psychological state of
the animal and can profoundly affect experimental responses (Baer, 1971; Clough, 1976).
Productivity, growth, and behaviour of laboratory mice may be seriously altered by
variations in floor space alone. Infant growth and survival, as well as maternal behaviour,

60
may be adversely affected by excessive floor space. Infant mortality in large cages can
occur from failure of females to nurse their young due to inhibition of mammary
development. Nest-building behaviour in rats is adversely affected in densely populated
pens, leading to an increasing tendency to ignore the pups and to infant death. Housing
density can affect efficiency of feed utilization and the incidence of skin lesions (Les,
1968, 1972).

Isolation stress may result in increases in nervousness, aggression, susceptibility to

convulsions and certain drugs, metabolism, and adrenocortical activity (Balazs and
Dairman, 1967; Hatch, Weiberg, Zawidzka et al. 1965; Moore, 1968). As much as
possible, housing type and densities should be kept uniform throughout a study. Further
details on appropriate housing (see also Laboratory Animal Facilities). Individual species
requirements are discussed in Volume 2 of this Guide (see also Social and Behavioural
Requirements of Experimental Animals). Recommended housing densities are listed in
Appendix I.

C. MICROBIOLOGICAL CONTROL

The effects that microbiological agents can have on experimental results and the health of
laboratory animals have been widely documented (Baker, Lindsey and Weisbroth, 1979;
Lindsey, Connor and Baker, 1978; Pakes, Lu and Meunier, 1984). Control of the
microbiological status of the experimental animal and its environment is necessary for
valid scientific results and animal well-being. The sources of microbial contamination
include vermin, experimentally infected and spontaneously ill laboratory animals or their
tissues or tumours, air, food, water, bedding, ancillary equipment, and personnel. Good
facility management practices and constant surveillance are necessary to minimize the
introduction of unwanted microbes. Insect and rodent vermin should be strictly controlled
or excluded from the facility (Small, 1983).

Whenever possible, the health status of all animals should be ascertained before the
animal is brought into the facility. Animals having an unknown health status should be
quarantined and tested before being admitted to the facility (Loew and Fox, 1983).
Additionally, all tumour and cell lines should be tested before being introduced (Small,
1984). Research on contagious diseases must be carried out in appropriate containment
facilities (see 3. below).

The laboratory animal veterinarian should be consulted about regular monitoring of the
health status of animals within a facility, as it is important to verify the microbiological
standing for publication of experimental results and to minimize cross-contamination
between areas (Baker, Lindsey and Weisbroth, 1979). The use of sentinel animals is one
proven, sensitive, and practical component of an animal health surveillance program
(Loew and Fox, 1983). Health monitoring programs should consider the source and
species of animal, husbandry practices, the nature of research carried out in the facility,
and the association of personnel with laboratory animals in other locations. The efficacy
of cage and equipment sanitation should be tested periodically by culturing for

61
microorganisms, as well as by checking physical indicators (Baker, Lindsey and
Weisbroth 1979; Small, 1983). Feed, water, and bedding should also be sampled and
cultured periodically. The frequency and intensity of microbiological monitoring
programs will be dependent upon husbandry practices, the level of confidence desired,
associated risk factors, and economics, in addition to the factors mentioned above (Small,
1984).

Personnel must be instructed in the precautions they must take to avoid introducing
diseases into the facility. The specific precautions will vary between areas and facilities,
depending upon the nature of the facility, the status of the animals, and the type of
research being conducted. The co-operation of all staff working with animals, in both
caretaking and experimental activities, is essential to maintain facility and scientific
standards.

1. Conventional Facilities

A conventional room or facility is one which is not especially designed for isolation
procedures. An isolation unit could operate conventionally if isolation management
practices are not employed. The following practices reduce the probability of
contamination in a conventional facility:

- Personnel should wear clean clothing and outer protective garments in animal rooms.

- Personnel should wash their hands upon entering and leaving a room.

- There should be no movement of personnel and equipment between rooms which

house animals of different
microbial status without proper precautions.

- Animals entering shared facilities, such as laboratories, surgery, irradiators, etc.,

should not be returned to
the holding room unless the shared room and equipment therein have been disinfected
between groups of
animals.

- Cleaning and sanitation practices as outlined in Laboratory Animal Care should be

followed.

2. Barrier Facilities

Gnotobiotes, SPF breeding colonies, aging study colonies, and immunodeficient or

immunosuppressed animals require a higher level of control of the microbial environment
than practised in conventional housing (Hessler and Moreland, 1984). Barrier housing
prevents infectious agents from entering and infecting animals inside the barrier. Barriers
can be established at the room level as in large-scale commercial production of disease-
free rodents; around groups of cages as for gnotobiotes or breeding colony nuclei in free-

62
standing flexible film isolators; or at the level of the individual cage as in microisolation
cages.

Closed barrier systems employ variations of the following principles:

- The room, isolator, or isolation cage is sterilized chemically or physically prior to

entry of animals, supplies,
or equipment.

- Animals enter through ports from isolators or transport containers which prevent
contamination.

- All other materials, supplies, and equipment are sterilized before entering the barrier.

- Effective entry and exit systems include pass-through autoclaves, sterilized double-
door transfer chambers,
or germicidal dunk tanks.

- Exits from large barriers may be through airlocks with powerful exhaust coming from
the inside of the unit.

- Personnel must shower, dress in sterile garments, and don head covers, masks and
Gloves before entering
a large barrier.

- The interior of smaller isolators is accessed through rubber or neoprene gloves sealed
to the isolation unit.

- Incoming air is filtered with high-efficiency particulate air (HEPA) filters and air
pressures are carefully
balanced to consistently prevent backflow into the barrier.

- Water is sterilized through filtration, UV light, acidification, or autoclaving.

- Feed and bedding are autoclaved or irradiated before entering the barrier. Special
enriched diets must be
used if the feed is to be autoclaved (Hessler and Moreland, 1984).

Microisolation cages are generally used to protect animals in otherwise conventional

rooms. With laminar flow cage-changing stations and special management procedures
(sterilization of feed, bedding, water, etc.), highly disease-susceptible animals such as
thymus deficient and Severe Combined Immune Deficiency (SCID) mice may be
successfully maintained in a conventional room. Rigorous microbiological monitoring is
necessary to maintain and verify the health status of animals kept in barrier-sustained
systems.

63
3. Biohazard Containment

Containment is required for animals exposed to known infectious microorganisms.

Required containment and management procedures vary with the biohazard classification
of the microorganism, based on the degree of risk to man and other animals (HWC/MRC,
1990). Personnel may be required to shower before leaving the containment unit. All
cages and materials are sterilized upon leaving the area. Air pressures are balanced so that
the highest pressure is outside the containment area. Air exiting the facility is diluted with
clean air, filtered, or incinerated. Because it is hazardous to staff and animals, UV light is
not generally recommended for routine disinfection of laboratory air. The infectious
disease unit should be segregated as much as possible from the rest of the animal facility.
Specific requirements will differ with the degree of risk. Depending on the hazard,
containment of small groups of animals may be accomplished with flexible film isolators
or microisolation cages. The use of laminar airflow racks to prevent cross-contamination
between cages should be carefully evaluated as the transfer of certain pathogens may be
enhanced in some instances (Clough, 1973). Infectious disease units should be
disinfected immediately following use. Recommendations for control of biohazards can
be found in Laboratory Biosafety Guidelines (HWC/MRC, 1990) and elsewhere (Barkley
and Richardson, 1984). Biological safety cabinets approved for the appropriate biohazard
level must be used for experimental manipulations. These cabinets must be inspected and
tested annually by trained personnel (HWC/MRC, 1990).

Persons working in infectious disease units should be protected with a comprehensive

occupational health and safety program.

D. CHEMICAL AND RADIOISOTOPE UNITS

In Canada, laboratory use of radioisotopes is regulated by the (federal) Atomic Energy

Control Board (AECB), in accordance with the Atomic Energy Control Regulations. The
AECB issues licences to the institution for the possession of radioactive material. When
radioisotopes are used in animals experimentally, Standard Operating Procedures (SOPs)
to ensure that related hazards are minimized should be defined and enforced; these SOPs
are considered by the AECB when it issues the Radiation Licence. As well, the AECB
recommends that the institution's Radiation Safety Officer sit on the Occupational Health
and Safety Committee in an ex-officio capacity.

The Workplace Hazardous Materials Information System (WHMIS) is regulated by

federal and provincial health and safety authorities. It legislates labelling requirements,
availability of Material Safety Data Sheets (MSDS), and training programs required for
personnel to work safely with certain hazardous materials.

The chemical and radiation hazard area should be separated from other animal housing
and work areas. The hazardous area must be clearly posted and entry restricted to
necessary personnel. Contaminated cages should not be transported through corridors.
Safe transport equipment and procedures should be developed if necessary. Laminar flow

64
cage-changing stations are recommended to protect the staff from aerosolized
contaminants (Hessler and Moreland, 1984) (see also Occupational Health and Safety).

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ALGERS, B. A note on behavioural responses of farm animals to ultrasound. Appl. Anim.

Behav. Sci. 1984; 12: 387-391.

ALLEN, D.M. A device providing gradual transitions between light and dark periods in
the animal room. Lab. Anim. Sci. 1980; 30: 252-254.

BAER, H. Long-term isolation stress and its effects on drug response in rodents. Lab.
Anim. Sci. 1971; 21: 341-349.

BAILEY, K.J., STEPHENS, D.B. and DELANEY, C.E. Observations on the effects of
vibration and noise on plasma ACTH and zinc levels, pregnancy and respiration rate in
the guinea pig. Lab. Anim. 1986; 20: 101-108.

BAKER, H.J., LINDSEY, J.R. and WEISBROTH, S.H. Housing to control research
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BALAZS, T. and DAIRMAN, W. Comparison of microsomal drug-metabolizing enzyme

systems in grouped and individually caged rats. Toxicol. Appl. Pharmacol. 1967; 10: 409-
410.

BARKLEY, W.E. and RICHARDSON, J.H. Control of biohazards associated with the
use of experimental animals. In: Fox, J.G., Cohen, B.J. and Loew, F.M., eds. Laboratory
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BELHORN, R.W. Lighting in the animal environment. Lab. Anim. Sci. 1980; 30: 440-
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BESCH, E.L. Environmental quality within animal facilities. Lab. Anim. Sci. 1980; 30:
385-406.

BRODERSON, J.R., LINDSEY, J.R. and CRAWFORD, J.E. The role of environmental
ammonia in respiratory mycoplasmosis of rats. Am. J. Path. 1976; 85: 115-127.

BUREK, J.D. and SCHWETZ, B.A. Considerations in the selection and use of chemicals
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CANADIAN COUNCIL ON ANIMAL CARE. Guide to the care and use of experimental
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CLOUGH, G., HILL, A. and BLACKMORE, D.K. Evaluation of a filter rack for
laboratory rodents. Lab. Anim. 1973; 7: 149-159.

CLOUGH, G. The immediate environment of the laboratory animal. In: McSheehy, T.,
ed. Control of the animal house environment. Laboratory animal handbook 7. Buckden
Huntington, Cambs., U.K.: Laboratory Animals Ltd., 1976: 77-94.

CLOUGH, G. Environmental factors in relation to the comfort and well-being of

laboratory rats and mice. Proc. of LASA-UFAW (Universities Federation for Animal
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CLOUGH, G. The animal house: design, equipment and environmental control. In:
Poole, T., ed. UFAW (Universities Federation for Animal Welfare) handbook on the care
and management of laboratory animals. 6th Ed. Harlow, Essex: Longman Scientific and
Technical, 1987.

CORNING, B.F. and LIPMAN, N.S. A comparison of rodent caging systems based on
microenvironmental parameters. Lab. Anim. Sci. 1992; 41: 498-503.

DAVIS, D.E. Social behavior in a laboratory environment. In: National Research Council
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FALK, S.A. and WOODS, N.F. Hospital noise levels and potential health hazards. New
Eng. J. Med. 1973; 289: 774-781.

FALL, M.W. The use of red light for handling wild rats. Lab. Anim. Sci. 1974; 24: 686-
687.

FLETCHER, J.L. Influence of noise on animals. In: McSheehy, T., ed. Control of the
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Laboratory Animals Ltd., 1976.

GAMBLE, M.R. and COUGH, G. Ammonia buildup in animal boxes and its effect on rat
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HATCH, A.M., WIBERG, G.S., ZAWIDZKA, Z., CANN, M., AIRTH, J.M. and GRICE,
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ITURRIAN, W.B. Effect of noise in the animal house on experimental seizures and
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KELLER, L.S.F., WHITE, W.J., SNIDER, M.T. and LANG, C.M. An evaluation of
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MCSHEEHY, T. Overview of the state-of-the-art in environmental monitoring. In: Melby,

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MELBY, E.C. JR. and BALK, M.W., eds. The importance of laboratory animal genetics,
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Chapter IV- Farm Animal Facilities and Environment

IV. FARM ANIMAL FACILITIES AND ENVIRONMENT

These guidelines are intended for farm animals used in agricultural research and teaching.
Where agricultural animal species serve as models for humans in biomedical research

69
projects and teaching demonstrations, they are to be kept in similar facilities compatible
with each animal's normal requirements and under conditions that will minimize stress,
bearing in mind the conditions required for non-agricultural species used in similar
experiments.

When farm animals are brought to the laboratory, consideration must be given to the
transition from the ambient outdoor conditions (e.g., cold weather, photoperiod), so that
the animals are given as smooth a transition period as possible. Bringing animals in from
the cold will result in physiological changes (e.g., hyperventilation in sheep) which will
also be reflected in changes in their dietary requirements. Husbandry procedures such as
shearing sheep, the trimming of hoofs, may also be of benefit to the animals at this time.
The time required for the animals to adapt to the laboratory's environment will vary.

The transition back to outdoor farm conditions following laboratory confinement also
requires careful planning, not only with respect to the ambient climate, but also with
respect to the regrouping of the animals.

Comprehensive guidelines for environmental enrichment, as well as for housing large

animals in metabolism crates, are found in the chapter on Social and Behavioural
Requirements of Experimental Animals.

The use of metabolism cages or crates necessarily reduces the animal's social and
behavioural activities. This practice should not, therefore, be used merely for the purpose
of convenient restraint, but should be reserved for approved metabolic studies. Animals
so housed should be under close and expert observation throughout the period of the
study (see also Social and Behavioural Requirements of Experimental Animals).

A. FACILITIES

Acceptable baseline information on facilities and housing for farm animals for production
purposes may be found in the National Research Council (NRC) Canadian Farm
Building Code (NRC, 1990). Similarly, the various recommended Codes of Practice for
livestock and poultry published by Agriculture Canada (Agriculture Canada, 1771/E,
1984; 1821/E, 1988; 1757/E, 1989; 1853/E, 1990; 1870/E, 1991) are also useful
references. In addition, a revision of the Recommended Code of Practice for the Care and
Handling of Farm Animals--Pigs (1898/E) is now in press.

Where a new facility or extensive remodelling of existing housing is contemplated, the

plans should be discussed with agricultural engineering experts (provincial departments
of agriculture and regional agricultural colleges). Detailed information is available in the
most recent edition of the Canadian Farm Building Code (NRC, 1990), and the
Canadian Farm Building Handbook (Agriculture Canada, 1988).

The American Guide for the Care and Use of Agricultural Animals in Agricultural
Research and Teaching (Curtis, 1988), contains useful information. The Scientists Center
for Animal Welfare (SCAW) has also published a volume on farm animal well-being

70
(Mench, Mayer and Krulisch, 1992).

A number of articles on Farm Animal Housing were compiled as a special feature by the
Veterinary Record (Wathes, Jones and Webster, 1983; Linklater and Watson, 1983;
Sainsbury, 1983). The British Veterinary Association's (BVA) Animal Welfare Foundation
has published guidelines for the detection and relief of pain in a number of species of
farm animals (Edwards, 1985; Gentle, 1985; Oldham, 1985; Silver, 1985). It has also
published guidelines on transportation of farm species (Gibson and Paterson, 1986).

Facility design and the nature of the primary enclosures used for the housing of farm
animals have a major impact on their welfare. The conditions for production-oriented
agricultural research must often be simulated and sometimes intensified in commercial
applications of intensive husbandry practices in food animal production (Fraser, 1975).
With others, however, attempting to impose close confinement can introduce a severe
stress and "skew" research results.

Probably the most important factor in the provision of appropriate animal care for farm
animals is the attitude and concern for animal well-being of the animal attendants and
herdsmen.

Domestication is a continuing process, and much of today's livestock and poultry

production involves animals of genetic strains that were selected for growth or
reproduction in various environments under varying degrees of control (Siegel, 1984).

Currently, no precise objective measures exist which can be employed to evaluate the
stress level of livestock production systems. Due to problems inherent in biochemical
monitoring, the physiological parameters of stress cannot be completely relied upon
(Freeman, 1971). The suggestion that high productivity does not constitute a reliable
indication of a lack of stress may, in some special instance, be correct. However, wide
acceptance of the negative correlation between stress and productivity has proven most
useful and beneficial in that its acceptance by agricultural producers has given rise to
continuing efforts to upgrade environmental conditions (Mann and Harvey, 1971; Wilson,
1971; Agriculture Canada, 1988). The Report of the Technical Committee to Enquire into
the Welfare of Animals kept under Intensive Livestock Husbandry Systems in Britain
concluded that no one factor can be considered conclusive in assessing well-being, and
the fact that farm animals are producing normally should be taken as no more than a
guide in this regard (Brambell, 1965). The well-being of farm animals probably will be
assessed best by an integrated system of indicators in four categories: 1) reproductive and
productive performance; 2) pathological and immunological traits; 3) physiological and
biochemical characteristics; and 4) behavioural patterns (Curtis, 1988; Duncan, 1981;
Curtis, 1982; Smidt, 1983).

Cages and pens should not only serve to confine the animal, but must also ensure its
comfort and safety by permitting normal postural and behavioural adjustments. Adequate
ventilation, ready access to food and water, and satisfactory viewing of the confined
animal are also mandatory. The Brambell Report in dealing with the implications of

71
modern technology on animal welfare, summarizes this concept by suggesting that
regardless of the system of management, five basic freedoms should be respected for all
farm animals; the freedom to get up, lie down, groom normally, turn around and stretch
its limbs (Brambell, 1965). Criticism that these criteria are not always fully met, and that
intensive livestock systems restrict living space and in some cases drastically reduce
freedom of movement, is often justified. The equivocal point is, to what extent the
potential stress of confinement is counterbalanced by such things as the period of the
imposed stress, injury prevention and improved disease control.

If slatted or partially slatted floors are used, the slat width and spacing will vary with the
species, but should be such as to provide adequate support and minimize the risk of injury
while permitting free drainage of excrement (Smith and Robertson, 1971). Slat material
should be durable. The possibility that toxic gases may develop from the liquid manure
disposal system must always receive consideration, as these may prove dangerous to both
livestock and personnel.

Solid floor surfaces for farm animals should be finished with materials and finishes that
will minimize slippage and thus the probability of injury and bruising. Epoxy resin floors
if properly keyed have been recommended for swine. The use of heavy rubber matting
(rubber cow mats) may prove useful in farrowing crates and for tethered animals, as well
as for stanchion-tied cattle. The arrangements for tethering animals in relation to each
other and to service areas within a facility may have a considerable influence on the well-
being, health and production of the animals. For example, sows in tie stalls will generally
thrive better if they can see each other and are fed simultaneously.

B. SPECIFIC ENVIRONMENTAL CONSIDERATIONS

Consideration of the facilities and environmental requirements of cattle and poultry have
been dealt with in more depth than have those for other classes of livestock. These two
species are used as type species in order to exemplify many of the principles common to
the environmental requirements of other species of farm mammals and birds.

1. Cattle

Conditions of housing for beef and dairy cattle suitable to northern hemisphere conditions
are well described in several Canadian and U.S. publications (NRC, 1990; Agriculture
Canada, 1988; Curtis, 1988; MWPS, 1987). It should be remembered that the
importance of good and intelligent management will increase proportionately with
the intensity of the animal production systems employed.

i) Temperature

Cattle are tolerant of a wide range of ambient temperatures provided they are healthy,
well-fed, and not exposed to extremes of solar radiation, humidity or high wind speeds
(Webster, 1983). Undesirable conditions of sanitation, mud, disease, parasites and various

72
insect pests reduce cattle tolerance to extremes of temperature.

Newborn calves are more vulnerable to extremes and fluctuations of temperature than are
older animals, with fluctuations tending to be more critical than the absolute temperature.
For dairy cows and calves maintained in closed housing systems, the optimum
temperature is one close to 20oC with an acceptable range between 10 and 25oC
(Sainsbury and Sainsbury, 1988).

Cattle maintained in free-stall and other open housing systems frequently choose to stand
in areas where the temperature is near or below 0oC. Cattle maintained in cold
environments require more total feed which can readily meet the extra maintenance
requirement of about 1% per each 1oC reduction in effective environmental temperature.
Under these conditions, productivity is not lowered and cattle do not appear to be
uncomfortable.

When still air temperatures climb above 25oC, feed intake and performance of heavily fed
cattle begin to be affected and they may become physiologically stressed. Tolerance to
heat and to cold vary with genotype. In general, beef cattle appear to be more winter
hardy than dairy cattle. In fact, the lower critical temperature of intensively fed and
housed dairy cattle is probably only about -7oC, while in beef cattle it can be -20oC.
Windbreaks in windy areas and overhead shelter in geographical areas subject to cold
rains, sleet and wet snow are highly desirable regardless of breed or type of cattle.

ii) Ventilation and humidity

The objective of a ventilation system is to provide the air exchange required to maintain
environmental temperature and humidity within the desired ranges and to remove
methane and carbon dioxide expelled from the rumen and lungs of cattle, ammonia from
the decomposition of feces and urine, dust from feed and bedding, and airborne
microorganisms.

In winter, removal of water vapour is a prime need in order to avoid condensation within
the building. Adequate insulation and, in some special cases, supplemental heat (e.g., calf
housing in certain locations) will also aid in maintenance of dry premises. Cold weather
ventilation rates should be sufficient to maintain relative humidity below 80% and above
40% (Curtis, 1983). During cold weather, ventilation in housing for neonatal animals
should maintain acceptable air quality without chilling the animals. In summer,
ventilation aids in keeping ambient temperature below the upper critical level of 25oC.
Ideally, the ventilation rate should be high enough to prevent indoor temperatures from
exceeding outdoor by more than 3oC when the atmospheric temperature is above 25oC
(Curtis, 1988).

A proper ventilation system should move the right amount of air for moisture and
pollutant removal in winter and for heat, moisture and pollutant removal in summer. The
system should provide a relatively uniform temperature, as this is more important than
the absolute temperature. Similarly, the airflow that is established through the building

73
should be even, so that neither drafts nor dead air pockets are created.

Open housing systems should be built so as to permit extra air movement in the summer
and minimum drafts in winter. Air current patterns are also of importance in winter in
relation to snow accumulation. Cattle should be able to feed, rest and exercise with
minimum exposure to cold wind and low temperature precipitation.

The indoor relative humidity range recommended in the Canadian Farm Building Code
is 25 to 75%. Levels of 50 to 55% humidity may be considered ideal and to provide a
minimum of influence on the physiological effects of other environmental parameters
such as temperature and ventilation. The comfort zone of animals is reduced (or
narrower) at both high and low temperatures under conditions of high humidity. High
humidity in closed housing at low temperatures leads to condensation; the resulting
dampness enhances the risk of disease transmission.

iii) Odours

Odours result from rumination by cattle, from feces and urine, from silage, spoiled feeds,
etc. Odours may taint the milk and, if highly repugnant to herdsmen, may also result in
the delivery of poorer animal care. Odours often indicate the presence of gases which can
be harmful to cattle and to man. This is especially true with modern liquid manure
systems in which hydrogen sulphide (H2S), ammonia (NH3) and methane (CH4) are
produced. High concentrations of H2S are lethal, and low to moderate levels of H2S and
NH3 are implicated in reduced animal health and performance. High CH4 concentrations
are explosive and at lower levels CH4 is a simple asphyxiant. CO2 results primarily from
rumen fermentation and exhalation by cattle. Except in extreme cases of poorly ventilated
housing, coupled with liquid manure agitation, CO2 accumulation is not considered to be
injurious to humans or animals. Occupational health standards for gases are shown in
Table 1.

TABLE 1 OCCUPATIONAL HEALTH STANDARDS FOR GASES

Gas TLVa ppm Excursion TWA
b
factor limitc ppm
H2S 10 2 20
NH3 25 1.5 37.5
CO2 5000 1.25 6250

a
TLV (threshold limit value) represents conditions under which it is believed that nearly
all workers may be repeatedly exposed for
an eight-hour day and 40 hour work week without adverse effect.
b
Excursion factor defines the magnitude of the permissible excursion about the TLV.
c
TWA (time-weighted average) limit defines the maximum concentration permitted for a

74
short exposure period.
TLV x Excursion Factor = TWA limit.

iv) Lighting

Light intensity must be adequate to maintain a high level of husbandry. For instance, an
intensity of 538 lux (50 fc) is desirable in the area of the udder in a milking parlour so
that the operator can properly care for the cow's udder. Two hundred and fifteen (215) lux
(20 fc) are quite adequate in most general housing situations with cattle. While
approximately equal hours of light and dark have generally been considered acceptable,
some evidence suggests that longer hours of light increases feed intake and cattle
performance.

v) Bedding

Bedding materials used in stalls and pens are chosen on the basis of availability, cost and
suitability, as well as the need. The housing system, and in particular the manure disposal
system, will largely dictate the bedding material if any, and how much is appropriate.
Straw or other appropriate materials are commonly used with or without rubber matting,
and on concrete, sand or wood bases. Comfort and cleanliness of animals is dependent
not only on amount and type of bedding, but also on animal stocking density, type of
shelter, temperature and humidity levels. In open housing under cold conditions, loose
straw is very helpful in minimizing heat loss from cattle, and a straw-manure base
sufficient to allow for fermentation can provide additional heat. Under conditions in
which cattle consume significant amounts of bedding, freedom from toxic compounds in
the bedding is critical.

vi) Population density

Conditions for housing of beef and dairy cattle are well described in the Canadian Farm
Building Handbook (Agriculture Canada, 1988). Space requirements vary depending on
size and type of animal, type of shelter, whether tied or loose, numbers of animals per
group, and level of management. As more intensive animal agriculture is practised,
quality of management must improve accordingly.

2. Sheep

General information on the facility requirements and environmental conditions suitable to

the raising and maintenance of sheep in Canada is available in a number of books and
monographs (Agriculture Canada, 1988; NRC, 1990; Curtis, 1988; Ensminger and
Parker, 1986).

i) Temperature

The comfort zone for the various classes of sheep has been reported as follows: ewes and

75
rams 7-24oC; feeder lambs 5-21oC; newly born lambs - until dry 24-27oC, which may be
provided by heat lamps (Ensminger and Parker, 1986). While these are stated to be the
comfort zones, sheep will not suffer in temperatures below -18oC if they are in fleece and
the humidity is low.

ii) Ventilation

The requirements vary widely due to geographic location. During the winter, the
recommended ventilating capacity of buildings housing sheep is 0.6-0.7 m3/min for each
ewe and 0.3 m3/min for each lamb. In summer the ventilating system should provide 1.1-
1.4 m3/min per ewe and 0.65 m3/min per lamb (Ensminger and Parker, 1986).

Preferred relative humidity is considered to be around 60%; however, a range from 50%
to 75% is acceptable (Ensminger and Parker, 1986).

iii) Lighting

There are no specific light requirements cited for sheep. Where windows equate to 3 to
5% or more of the floor area, these will provide sufficient natural light. Photoperiod may
need to be regulated for the purpose of controlling the onset of estrus.

Although natural light is normally sufficient for sheep in most situations, supplemental
lighting should be provided during lambing periods.

iv) Bedding

Straw is the most common bedding material used. Some modern units use a liquid
manure system with floors of expanded metal, wire or slats and no bedding. Providing
that space allocation is correct, these systems are acceptable.

v) Population density

Table 2 taken from the 1988 edition of the Canadian Farm Building Handbook
(Agriculture Canada, 1988) gives the accepted detail for sheep accommodation. The
space allocation cited may be considered generous from the viewpoint of practical
commercial sheep raising, but acceptable for research and teaching animals.

Generally the number of animals per pen should not greatly exceed 100 pregnant ewes or
50 ewes with lambs or 500 feeder lambs.

TABLE 2 ACCOMMODATION FOR SHEEP

ACCOMMODATION EWES AND RAMS FEEDER LAMBS
Feed lot (m2/head) 1.4 0.6
hard surfaced 6.5 2.8
soil1

76
 Open-front shed floor area (m2) 1.4 0.6
Pregnant ewe 0.93
Dry ewe 2.7 2.7
Ceiling height minimum (m)
Slotted floors (m2/head)2 0.65 0.4
% floor area slotted 100 100
slot width (mm) 19 16
slat width 50-75 50-75
Lambing pen, not slotted (m, 1.2 x 1.2
minimum) 1.2 x 1.5
Claiming pen only
Lambing and claiming pen
Feed rack, length per head (mm) 400 300
Group feeding 150 100
Self-feeding

Height at throat (mm) 300 250

Small breeds 375 300
Large breeds
Feed storage 1.4 0.9
Hay (kg/day per head) 2.3
Small breeds
Large breeds 0.15 0.23
Grain (kg/day per head) 0.45-1.13
(maintenance)
(finishing)
Bedding storage (kg/day per head) 0.34 0.11
2
Water surface area (m /40 head) 0.1 0.1

1
Soil-surfaced feed lots should be used only where annual precipitation is less than 500
mm. A paved strip next to the feed bunk
should be at least 1.8 m wide, or as wide as the tractor used for cleaning. The strip
should slope 1:25 away from the feed bunk.
2
An alternative to slotted floors, for ewes, rams or lambs is 25 x 50 mm, 4 mm-gauge
expanded and lattened metal mesh.
Expanded metal mesh floors may be covered with a solid panel to retain bedding for
lambing.

3. Swine

Detailed information and guidelines for swine housing may be found in the Canadian
Farm Building Handbook (Agriculture Canada, 1988). The Veterinary Infectious Disease

77
Organization has published three booklets on Farrowing Barn Design and Management,
Swine Nursery Design, and Feeder Barn Design and Management, designed to provide
swine producers with current information on modern building design and operation
(VIDO, 1986, 1987).

The internal surfaces of all swine houses and equipment contained therein should be
constructed of smooth, non-porous materials which can be readily and effectively cleaned
and disinfected. Pen dividers and feeders should be free of sharp edges or projections
which might cause injury to the animals. Passageway and pen floors should be effectively
drained. All floors, whether solid, slatted, or wire mesh, should provide adequate footing
and be non-injurious to the pigs.

It is not feasible to state specific values for such environmental parameters as

temperature, humidity and ventilation that are meaningful for all classes of swine in all
possible research and teaching situations. The precise requirements will vary
considerably with age, type of housing, density of population, etc., and the ranges cited in
most instances refer to the upper and lower limits of generally accepted comfort zones.

i) Temperature

With the possible exception of the neonatal and nursing piglets, swine are extremely
adaptable and comfortable over a wide range of climatic conditions, if they are provided
with the proper facilities to conserve or dissipate body heat. Pole barns or outside huts
can be comfortable even in extremely cold weather, if the unit has a sufficient population
and is provided with adequate and appropriate bedding for the pigs to create a
comfortable microenvironment. Animals with access to outside runs or paddocks in hot
weather should have a shaded, preferably damp, area so they can stretch out on the
ground and dissipate body heat by conduction. Total confinement, on concrete or slats,
may interfere with conductive heat transfer so the environmental support systems must be
adequate to maintain a satisfactory comfort zone through all seasons.

For adults and most growing pigs (>30 kg) the comfort zone range is about 15-25oC
(Curtis, 1988). The farrowing facility presents a special concern because the
environmental requirements for the sow, and the newborn piglet are drastically different.
For the comfort of the sow a temperature of 15-26oC should be maintained, whilst the
creep area should be dry, draft-free and provide a temperature of from 26-32oC at all
times for the newborn piglets (Curtis, 1988).

ii) Ventilation and humidity

Adult and growing pigs will thrive at a relative humidity within the range of 40-80%
(Curtis, 1988). Ventilation rates in winter should be sufficient to control moisture. In
summer the airflow rates required to remove heat produced by the animals are 15-20 X
higher than the rates required for moisture control (VIDO, 1987). Metal bars or wire
mesh partitions between individual pens are preferable to solid structures as they
facilitate air movement at the level of the animal.

78
iii) Lighting

Photoperiod has a definite effect on the age at which sexual maturity is achieved and may
also influence growth rate and feed efficiency (Maybry, Jones and Seerley, 1983),
although Berger, Mahone, Svoboda et al. (1980) suggest that no particular photoperiod is
necessary for growing pigs. From the viewpoint of good animal care, the light intensity
should be such that animals in all areas of the facility can be observed clearly at all times.

iv) Noise and odours

Some noise and odour will inevitably be present in any practical swine unit. Odours may
be minimized by regular efficient cleaning and adequate ventilation. Noise levels can be
held down by ensuring that mechanical equipment operates relatively quietly and by
minimizing procedures which disturb the animals.

v) Bedding

Where swine are held for relatively short periods in small units, straw or other
appropriate materials may be used. In large units, with automated cleaning and manure
handling, it is customary to house pigs without bedding. Where pigs are maintained in
pole barns or outside huts, deep straw bedding should be provided.

vi) Population density

Young pigs up to 10 or 12 weeks of age get along very well, and substantial numbers can
be kept together in a single pen (Curtis, 1988). As the animals grow older, aggressive
behaviour develops and fighting or bullying will occur, particularly in larger groups.
Generally, group size should be kept to ten or less for mature sows and animals in the late
growing phase (Sainsbury and Sainsbury, 1988). With electronic feeding stations, larger
groups may be more appropriate. Whenever groups of sows are first established, some
fighting will take place for the first few days. The belligerent interactions between
individuals within the group should subside as the social hierarchy is established;
however, it is difficult to add new animals to previously established groups. If the
grouped pigs are limit-fed, it is essential that sufficient feeder space be provided so that
all animals can eat at the same time.

When adult pigs are confined to stalls, the stall should always be long enough to allow
the pig to lie fully relaxed without its head or nose touching the feeder or front of stall.
The stall must also be wide enough to allow the animal to lie fully relaxed on its side with
its feet and legs extended. A stall width of 0.65 m will usually satisfy this requirement.

Although not recommended by the Canadian Council on Animal Care (CCAC), if a tie
stall is to be used, extreme care must be taken with regard to the design of the collar or
belt utilized and the tethered pig(s) must be closely monitored when first restrained. If

79
any abrasions occur in the region of the collar or belt the animal must be released
immediately. As a general rule, the tie system should not be used unless the animal
has been acclimatized to it at an early age. Where slatted or partially slatted floors are
used, care must be exercised to insure that slot width is such that no portion of the pig's
hoof or leg will pass through. Particular attention must be paid to the floor structure when
dealing with newborn piglets. Tables 3 and 4 are pen floor space allowances for growing
pigs and replacement gilts and sows which will be included in a forthcoming
Recommended Code of Practice for the Care and Handling of Farm Animals--Pigs
(Agriculture Canada, 1898/E in press).

TABLE 3 RECOMMENDED PEN FLOOR SPACE ALLOWANCES FOR GROWING

PIGS BASED ON BODY WEIGHT.667
Body Weight Fully Slatted Partial Slats Solid Bedded
.667 .667
(0.035*BW )++ (0.039*BW ) (0.045*BW.667)
kg (lbs) m2 (ft2) m2 (ft2) m2 (ft2)
10 22 0.16 (1.7) 0.18 (1.9) 0.21 (2.2)
20 44 0.26 (2.8) 0.29 (3.1) 0.33 (3.5)
50 110 0.48 (5.2) 0.53 (5.7) 0.61 (6.6)
75 165 0.62 (6.7) 0.70 (7.5) 0.80 (8.6)
90 198 0.70 (7.5) 0.78 (8.4) 0.91 (9.7)
100 220 0.76 (8.2) 0.85 (9.1) 0.97 (10.4)
110 242 0.81 (8.7) 0.90 (9.7) 1.03 (11.1)
++For calculations; body weight (BW) is in kg, area in m2.

TABLE 4 RECOMMENDED PEN FLOOR SPACE ALLOWANCES FOR

REPLACEMENT GILTS AND SOWS
Body Weight Partial Slats Solid Bedded
.667
(0.054*BW )++ (0.059*BW.667)
kg (lb) m2 (ft2) m2 (ft2)
100-150 (220-330) 1.5 (16) 1.7 (18)
150-200 (330-440) 1.8 (19) 2.0 (22)
200-250 (440-550) 2.1 (23) 2.3 (25)
>250 (>550) 2.3 (25) 2.6 (28)

++For calculations; body weight (BW) is in kg, area in m2.

4. Horses

The basic conditions for housing and maintaining a proper environment for horses are

80
outlined in the Consortium Guide (Curtis, 1988) and also in the Horse Housing and
Equipment Handbooks (MWPS, 1986). The Canadian Farm Building Handbook
(Agriculture Canada, 1988) deals specifically with housing for riding horses.

A bright, airy stable with access to an exercise paddock is desirable in housing horses in
order to maintain top condition, muscle tone and health. The housing area should allow
for adequate space within its alleyways to permit the safe movement of horses and
attendants.

Stall construction should preferably be of hardwood at least 3.75 cm thick. Doors and
partitions should be metal faced, particularly along their top edges, to discourage
"cribbing." There should be no protuberances which might be injurious to the animals.
Stall walls should be of sufficient height to prevent interference with adjacent animals. It
is important that doors be of adequate width (1.25 m) and height (2.25 m) to permit the
easy movement of horses, without risk of injury. Ceilings and overhead supporting beams
should also be of sufficient height, preferably 3 m, to permit the horse to assume a normal
posture and to guard against possible head injury.

Floors should have durable, non-slip surfaces. Roughened concrete is satisfactory; wood
flooring in the standing stall is often used; packed earth may, in some instances, be
acceptable in box stalls. Under institutional and laboratory conditions, a room or area
separate from the stable area should be provided to perform special procedures (e.g., the
collection of large quantities of blood, etc.).

i) Temperature

Horses can tolerate low temperatures provided there is adequate shelter from extremes of
wind, rain and snow. Similarly, quite high temperatures can be tolerated provided
adequate shade is available to the free-ranging animal and that appropriate ventilation and
humidity are provided in the stable. An abundant supply of fresh, potable water should be
available at all times, and is particularly important in hot weather. When housed in a dry
draft-free environment, horses can also tolerate a wide range of environmental
temperatures (-7 to 29oC). However, the optimum appears to lie between 10 and 15oC
(Ensminger, 1969). The relative humidity in horse quarters should range between 50-80%
(Curtis, 1988).

ii) Ventilation

Ventilation rate capacity should be at least 0.7 m3/min. per 450 kg of horse at
temperatures of -18 to -7oC and 2.8 m3/min. per 450 kg of horse at temperatures of -1 to
10oC (MWPS, 1987). The capacity will need to be increased during hot weather.

iii) Lighting

The level of lighting in a horse barn should allow for adequate examination of animals
and bedding. Total darkness should be avoided. A light source should be present at night.

81
Illumination of at least 200 lux is recommended for alleys, handling and feeding areas
(Currence and McFate, 1984). One 100 W incandescent lamp per 8 m of floor or each
box stall will produce the required light level (MWPS, 1987).

iv) Bedding

Sufficient bedding should be provided in the form of straw, wood shavings or other
suitable material. Adequate floor drainage must be assured in both the box and standing
stall to guard against foot problems and unnecessary soiling of the animal.

Manure and soiled bedding should be removed daily to keep horses clean and dry, and the
environment free of dust and odours (Curtis, 1988). The animal should not have access to
the manure storage area due to the high risk of parasite infestation from this source.

Regular grooming is strongly advocated for members of this species, particularly if

housed in tie stalls with limited freedom of movement.

v) Population density

Ideally juvenile and mature horses should be housed in individual box stalls of at least 3.5
m x 3.5 m, with access to an exercise paddock of 10.0 m x 27.50 m or larger. Horses may
be quite satisfactorily maintained in standing stalls providing there is good separation
between each animal and regular access to an exercise paddock. Subsequent to weaning,
it is desirable that each animal have a separate stall. In the exercise paddock, separation
by age and compatibility should be maintained.

5. Poultry

It is not feasible in the space of a few pages to deal in detail with the housing, feeding and
management of poultry. More detailed information may be found in the literature (Curtis,
1988; Agriculture Canada, 1988; Moreng and Evans, 1985; North and Bell, 1990).

a) Chicks

In experimental studies, chicks are either brooded and reared in floor pens or in batteries.
Buildings that house such facilities should be designed and operated in such a manner as
to provide maximum comfort for the birds and minimum risk of disease transmission.

It is customary for such buildings to have concrete floors equipped with floor drains and
walls of sealed concrete block or sealed plywood to facilitate easy cleaning and
disinfection. Insulation of walls and ceilings is essential as is adequate ventilation.

i) Temperature

Initial brooding temperature should be 35oC as measured on a level with the backs of the
chicks. As the birds age, the brooding temperature should be reduced at the rate of about

82
2.5oC per week. By the time the birds are 5-6 weeks old, the house temperature should be
down to 18 to 21oC (Curtis, 1988). A thermometer alone, however, is a poor tool for
ensuring chick comfort. The chicks themselves should also be the indicators (North and
Bell, 1990).

ii) Ventilation

All poultry buildings must be adequately ventilated either naturally or by forced air. In
most installations, the minimum ventilation rate in summer should be about 12 air
changes per hour. Such ventilation rates are usually adequate to keep ammonia levels in
the buildings down to acceptable levels. Levels of ammonia in poultry houses should not
exceed 25 ppm. Higher levels are likely to prove detrimental to the birds and
uncomfortable for attendants.

iii) Lighting

Lighting systems vary widely; however, artificial light controlled by a time clock should
always be employed. Chicks brooded in batteries are usually subjected to about 35 lux
(3.5 fc) of white light on a continuous basis for the first four days after hatching. Broiler
chicks brooded in floor pens should receive 35 lux of illumination at the floor for the first
48 hours after hatching, with a light cycle lasting 23 hours, the dark cycle one hour. A 23-
hour program is preferable to 24 hours of light, because it acquaints the flock with
periods of darkness. This is very bright illumination; however, it is essential that chicks
learn to drink and eat as soon as possible. After the first two days, light intensity should
be reduced to about 10 lux (1 fc) at floor level (North and Bell, 1990). Replacement stock
are generally lighted like broiler chicks until about six weeks of age when a restricted
lighting schedule is introduced. Such schedules, over a period of time, reduce the hours
of light to about eight hours per day.

iv) Bedding

Many different types of litter have been used successfully in floor pens. Preference in
most parts of Canada is for wheat straw or wood shavings. In general, visual cleanliness
of litter is considered of less importance than dryness; spillage of water must be
minimized by using suitable waterers. A sufficient number of air changes must be
provided in the building to remove the moisture-laden air from the pens; at the same time,
care must be taken not to reduce the moisture level to the point where a dust problem is
created.

v) Food and water

A good supply of fresh, clean water must be provided and maintained at all times.
Feeders of many different types (troughs, hanging feeders and mechanical feeders) have
all been used successfully. Where practical, they should be provided with reels or grills to
prevent wastage and fouling of the feed by the birds. Feeders should be of a type and size
suitable for the age (size) of the birds. Sufficient feeder space must be provided to permit

83
all of the birds to eat at one time.

vi) Population density

Some guidelines for space requirements for chickens will be found in Table 5. The
recommended allowances should be considered as "rules of thumb" rather than as
absolute minimum allowances.

TABLE 5 GUIDELINES FOR MINIMUM SPACE REQUIREMENTS FOR POULTRY

Floor area/bird
cm2
Brooding and Growing Period 64a-1116b
Floor pen housing 0-6 weeks 742a-2786b
>6 weeks

Cage housingc Leghorn-type/Medium- 97/155

size 194/310
0-6 weeks 290/348
6-12 weeks
12-20 weeks
Laying Period 1625
Floor pen housing on litter 1858
Leghorn-type
Medium-size
387
Cage housing 452
Leghorn-type
Medium-size
Breeding flocks - males and females 1393a-2786d
Floor pen housing on litter
Feeder space - Length/bird cm. 10
Nests - per 100 layers 25

Adapted from Curtis (1988) and North and Bell (1990)

a
Mini-Leghorn pullets
b
Meat-type cockerels
c
Cages should allow birds to stand erect
d
Meat-type

b) Laying and Breeding Hens

In general, the type of building required for housing laying and breeding chickens is
similar to that required for floor brooding of chickens; however, the internal design and

84
equipment required are different. Houses for layers and breeders maintained on floors are
usually equipped with dropping pits or dropping boards, nests, appropriate troughs or
hanging feeders, and automatic waterers.

In order to prevent cannibalism, the birds are often beak-trimmed with an electric beak
trimmer. Heavily beak-trimmed birds may suffer a severe setback in growth and in
subsequent performance in the laying house. Excessive trimming should, therefore,
always be avoided both on humane and economic grounds.

i) Ventilation

Ventilation rates required in summer and winter in houses for laying and breeding
chickens are fairly similar to those required in houses for rearing replacement stock. In
winter, the system must remove moisture build up whilst maintaining an optimum house
temperature between 18 and 24oC. In the summer, the ventilation system should maintain
the house temperature below 27oC. At temperatures above 27oC laying pullets begin to
suffer and performance diminishes (North and Bell, 1990).

ii) Lighting

Artificial light, controlled by a time clock, must be provided to layers for optimal
production. It is usual to provide 14 hours of white light per day at a light intensity of 10
lux (1 fc) at the feeders and waterers (North and Bell, 1990).

iii) Bedding

The floors are usually covered with litter of straw or shavings. Occasionally, such houses
have floors of 2.5 cm x 5 cm mesh, heavy gauge, electrically welded wire which obviates
the need for litter.

Houses suitable for maintaining layers in cages usually have single or multiple cages
arranged in rows about 76 cm apart. Stair-step or single-deck type cages, with mechanical
cleaning arrangements underneath are perhaps the most popular cages, since removal of
droppings is made easier with such cages than with multiple-deck cages.

iv) Food and water

Feeds appropriate to the stage and level of production are readily available commercially.
Food may be supplied in mash or pelleted form. Laying or breeding rations usually
contain about 16% protein. Water is generally supplied by automatic waterers. Both
trough and large cup waterers have been found satisfactory for layers in floor pens while
trough, drip and small cup waterers have been found suitable for layers in batteries.

v) Population density

Some guidelines for space requirements for hens will be found in Table 5.

85
c) Commercial vs. Experimental Conditions

The housing, feeding and management of chickens have been treated above as in
commercial practice. The principal difference between the commercial situation and the
experimental situation is that in the latter, many different treatments and replications are
involved which often may necessitate the use of many small floor pens or many small
battery groups. These must be group- or individually-fed with individual or group data
being collected from each. Production-oriented agricultural research, in order to be
relevant, will most often require an approximation of good commercial management and
housing practices.

Where chickens are utilized as a bioassay tool in biomedical and behavioural research,
the environmental conditions given above for chicks and mature birds should be
considered as constituting minimal acceptable standards. When chickens must be
introduced into a laboratory animal facility in which poultry are not usually
accommodated, it is necessary to provide appropriate caging. In these circumstances,
advice should be sought from someone knowledgeable in poultry science and husbandry
as well as laboratory animal science to assure that such considerations as sufficient head
room, feeder space and proper flooring are satisfactorily met. Care should also be taken
to ensure that feeders and waterers (particularly where fountains and open cups are used)
are located so as to avoid becoming fouled with feces or clogged with bedding.

C. PEST CONTROL

Programs should be in place to control infestation by vermin (flies, mosquitoes, fleas,

lice, ticks, rodents, skunks and birds). The most effective control is by preventing entry
by the appropriate screening of openings and sealing cracks, maintaining the integrity of
all surfaces, and eliminating vermin breeding sites. Pesticides should only be used
judiciously and when necessary and where the risk to animals and the experimental
process is minimal.

Cats are sometimes used for rodent and bird control and if so they should receive
appropriate veterinary care including complete inoculation against the common feline
diseases, including rabies.

D. REFERENCES

AGRICULTURE CANADA. Publication 1771/E. Recommended code of practice for the

care and handling of pigs. Communications Branch, Agriculture Canada, Ottawa, Ont.,
K1A 0C7. 1984.

IBID. Publication 1898/E. Recommended code of practice for the care and handling of
farm animals--pigs. In Press.

86
IBID. Publication 1821/E. Recommended code of practice for the care and handling of
special fed veal calves. 1988.

IBID. Publication 1757/E. Recommended code of practice for the care and handling of
poultry from hatchery to processing plant. 1989.

IBID. Publication 1853/E. Recommended code of practice for the care and handling of
dairy cattle. 1990.

IBID. Publication 1870/E. Recommended code of practice for the care and handling of
farm animals--beef cattle. 1991.

AGRICULTURE CANADA. Publication 1822/E. Canadian farm building handbook.

Communications Branch, Agriculture Canada, Ottawa, Ont., K1A 0C7. 1988.

BERGER, T., MAHONE, J.P., SVOBODA, G.S., METZ, K.W. and CLEGG, E.D. Sexual
maturation in boars and growth of swine exposed to extended photoperiod during
decreasing natural photoperiod. J. Anim. Sci. 1980; 51: 672-678.

BRAMBELL, F.W.R. CHR. Report of the Technical Committee to Enquire into the
Welfare of Animals Kept Under Intensive Livestock Husbandry Systems. Her Majesty's
Stationery Office, London, England 1965.

CURTIS, S.E. Measurement of stress in animals. In: Woods, W.R., ed. Proc. Symp.
Manage. Food Producing Anim. Purdue Univ., West Lafayette, IA, 1982; 1: 1-10.

CURTIS, S.E. Environmental management in animal agriculture. Ames, IA: Iowa State
Univ. Press, 1983.

CURTIS, S.E., ed. Guide for the care and use of agricultural animals in agricultural
research and teaching. Consortium for Developing a Guide for the Care and Use of
Agricultural Animals in Agricultural Research and Teaching (309 West Clark Street,
Champaign, IL 61820) 1988.

CURRENCE, H.D. and MCFATE, K.L. Agricultural wiring handbook. 7th Ed. Publ.
8401, Columbia, MO: Natl. Food and Energy Counc., 1984.

DUNCAN, I.J.H. Animal rights - animal welfare: A scientist's assessment. Poult. Sci.
1981; 60: 489-499.

EDWARDS, G.B. Clinical assessment of pain, distress and discomfort in ruminants. In:
Gibson T.E. and Paterson, D.A., eds. The detection and relief of pain in animals. London:
British Veterinary Association Animal Welfare Foundation, 1985: 85-88.

ENSMINGER, M.E. Horses and Horsemanship. 4th Ed. Danville, IL. The Interstate

87
Printers and Publishers Inc., 1969.

ENSMINGER, M.E. and PARKER, R.O. Sheep and goat science. 5th Ed. Danville, IL.
The Interstate Printers and Publishers Inc., 1986.

FRASER, A.F. Behaviour of livestock under intensive conditions of husbandry. Appl.

Anim. Ethol. 1975; 1: 111-112.

FREEMAN, B.M. Stress and the domestic fowl; a physiological appraisal. World Poultry
Sci. J. 1971; 27: 263-275.

GENTLE, M. Measurements of pain, distress and discomfort in poultry and other birds.
In: Gibson T.E. and Paterson, D.A., eds. The detection and relief of pain in animals.
London: British Veterinary Association Animal Welfare Foundation, 1985: 92-95.

GIBSON, T.E. and PATERSON, D.A., eds. The welfare of animals in transit. London:
British Veterinary Association Animal Welfare Foundation, 1986.

LINKLATER, K.A. and WATSON, G.A.L. Sheep housing and health. Vet. Rec. 1983;
113(24): 560-564.

MAYBRY, J.W., JONES, R.D. and SEERLEY, R.W. A comparison of an 8-versus 16-
hour photoperiod during lactation on suckling frequency of the baby pig and maternal
performance of the sow. J. Anim. Sci. 1983; 57: 292-295.

MANN, C.M. and HARVEY, P.N. Cage size and stocking density for laying hens. World
Poultry Sci. J. 1971; 27: 350-356.

MENCH, J.A., MAYER, S.J. and KRULISCH, L. eds. The well-being of agricultural
animals in biomedical research. Bethesda, MD: SCAW (Scientists Center for Animal
Welfare), 1992.

MIDWEST PLAN SERVICE. Horse housing and equipment handbook. 7th Ed. Ames,
IA: MidWest Plan Service, Iowa State University, 1986.

IBID. Structures and environment handbook. 11th Ed. Ames, IA: MidWest Plan Service,
Iowa State University, 1987.

MORENG, R.E. and EVENS, J.S. Poultry science and production. Reston, VA. Reston
Publishing Company, Inc., 1985.

NATIONAL RESEARCH COUNCIL. Canadian farm building code. 7th Ed. Ottawa,
Ont.: National Research Council, 1990.

NORTH, M.O. and BELL, D.D. Commercial chicken production manual. 4th Ed. New
York, NY: Van Nostrand Reinhold, 1990.

88
OLDHAM, J.G. Clinical measurement of pain, distress and discomfort in pigs. In:
Gibson T.E. and Paterson, D.A., eds. The detection and relief of pain in animals. London:
British Veterinary Association Animal Welfare Foundation, 1985: 89-91.

SAINSBURY, D.W.B. Poultry housing and disease. Vet. Rec. 1983; 113(24): 565-568.

SAINSBURY, D.W.B. and SAINSBURY, P. Livestock health and housing. 3rd Ed.
Toronto, Ont.: Baillire Tindall, 1988.

SIEGEL, P.B. The role of behaviour in poultry production: A review of research. Appl.
Anim. Ethol. 1984; 11: 299-316.

SILVER, I.A. Horses. In: Gibson T.E. and Paterson, D.A., eds. The detection and relief of
pain in animals. London: British Veterinary Association Animal Welfare Foundation,
1985: 82-84.

SMIDT, D. Advantages and problems of using integrated systems of indicators as

compared to single traits. In: Smidt, D., ed. Indicators relevant to farm animal welfare.
Boston, MA: Martinus Nijhoff, 1983: 201-207.

SMITH, W.K. and ROBERTSON, A.M. Observations on injuries to sows confined in part
slatted stalls. Vet. Rec. 1971; 89: 531-533.

VETERINARY INFECTIOUS DISEASE ORGANIZATION. Farrowing barn design and

management. VIDO (124 Veterinary Road, Saskatoon, Saskatchewan S7N 0W0), 1986.

IBID. Swine nursery design; feeder barn design and management 1987.

WATHES, C.M., JONES, C.D.R. and WEBSTER, A.J.F. Ventilation, air hygiene and
animal health. Vet. Rec. 1983; 113(24): 554-559.

WEBSTER, A.J.F. Environmental stress and the physiology, performance and health of
ruminants. J. Anim. Sci. 1983; 57: 1584-1593.

WILSON, W.O. Evaluation of stressor agents in domestic animals. J. Anim. Sci. 1971;
32: 578-583.

89
 Chapter V - Laboratory Animal Care

V. LABORATORY ANIMAL CARE

A. INTRODUCTION

a) Animal Care and Handling

All animal facilities should have in place Standard Operating Procedures (SOP) for
animal care. Assistance in this regard may be found in a manual and computer program
developed by Olson, Morck, and Nabrotzky (1992).

All animals must be observed at least once daily.

Animals may have to be handled when being put into new cages or removed for various
experimental purposes. Most domestic and laboratory animals need no restraint for such
routine handling, and will respond to gentleness. Under normal conditions, all standard
laboratory animals, except non-human primates (NHP), can be handled without the use of
gloves or other restraining aids. In all cases, only the minimal amount of force necessary
should be employed. Manipulation of the type and intensity of light used often proves
useful in handling small wild mammals and birds (Fall, 1974).

Successful handling requires the ability to recognize the animal's state of mind, which
may include bewilderment, apprehension and, in some cases, discomfort or pain. Proper
training is important to provide consistency in handling which usually results in more
manageable animals.

Whenever possible, the use of cumbersome protective garments, such as gloves, should
be avoided, as they often prevent the handler from developing the proper sense of touch
and cause the animal discomfort. However, with wild and semi-domesticated species
(mink, monkeys, etc.) use of protective gauntlets and restraining equipment is usually
necessary. The use of specialized cage squeeze mechanisms followed by tranquillization
is often advocated for handling the larger NHP. Transfer devices, pole tether devices and
training through reward may be used to effect routine cage changes. Transfer devices
have also been designed and utilized for a variety of small wild rodent species (Caudill
and Gaddis, 1973).

The handling of individual species is addressed in Volume 2 of the Guide. In addition,

Agriculture Canada has published Codes of Practice for handling many species of farm
animals (Agriculture Canada, 1771/E, 1984; 1821/E, 1988; 1757/E, 1989; 1853/E, 1990;
1870/E, 1991). In addition, a revision of the Recommended Code of Practice for the Care
and Handling of Farm Animals--Pigs (1898/E) is now in press.

90
B. GENERAL PRACTICES

1. Reception

The portal through which animals and containers enter a facility is important in the
overall prevention of disease. The examination of newly arrived animals should have
several aims: evaluating the condition and health of the animals; preventing cross-
contamination of animals from different sources; and ensuring that the order has been
accurately filled. The health status of the animals at their source and the possibility of
cross-contamination during transport are important considerations. Cross-contamination
is always a greater risk if the animals are not shipped in a vehicle dedicated to the
transport of animals, coming from a single source. However, this risk can be decreased by
the use of filtered shipping crates.

Each new shipment of animals should be received, uncrated and examined by trained
personnel and placed in clean cages in a designated reception area separate from the
animal holding room(s). The reception area should be cleaned and disinfected after each
new shipment. Shipping containers should not enter the main facility unless properly
decontaminated and should be properly disposed of, or thoroughly cleaned and
disinfected if they are to be reused. Incoming animals should be identified and their
arrival appropriately recorded.

Animals that appear unhealthy, or which have been in any way debilitated in transit,
should be separated from the remainder and held in an appropriate location for
observation and treatment. Where this is not feasible, such animals should be euthanized
without delay.

2. Conditioning/Quarantine

The level of conditioning required will depend on the differences in microbial status
between the resident animals and the incoming animals. Rodents are commonly
purposebred for the laboratory and can be received from the supplier in a defined state,
with a known health, nutritional and, to a varying extent, genetic background. Similarly
other purposebred species obtained from reputable sources will have complete health
profiles and will have received specified prophylactic treatment. Such animals may not
normally require further quarantine for confirmation of their health status; however, a
holding period of several days will give the animals the opportunity to adjust to their new
surroundings. A minimum adjustment period of two days is required after shipping
for immune function, corticosterone levels and other physiological parameters to
stabilize (Small, 1984; Toth and January, 1990).

Legally procured strays or donated animals, acquired feral animals, such as NHP, and
animals from other random sources should be subjected to a period of conditioning
following their reception. Conditioning requires that the animal be held for a varying
period of time (1-6 weeks) in separate quarters. The length of the conditioning period will

91
depend on the species, the health status of the animals, the reliability of the supplier, and
whether an active process of screening animals for the presence of or exposure to
infectious agents is undertaken. During this period, thorough physical examinations
should be undertaken. Further examination will depend on species and intended use.

The conditioning period should be of sufficient duration to permit the proper evaluation
of the suitability of the animal(s) for their intended use, and testing for contagious,
zoonotic and other diseases that may be of concern. This can include serological
screening for antibodies to viruses and other pathogens, the examination for ecto- and
endo-parasites, mycoplasma, and pathogenic bacteria. If contamination during transport
is a possibility, an appropriate period to allow expression of disease or antibody
production should be allowed. Sufficient time must also be allowed for appropriate
treatment or vaccination against diseases that tend to be endemic in the species being
conditioned.

During the conditioning/quarantine period, the animals should be held in facilities

separated from other animals with no crossover of personnel, equipment, supplies or
ventilation (Frost and Hamm Jr., 1990), unless effective measures are taken to prevent
cross-contamination.

3. Holding (Maintenance)

Only one species should be housed in a conventional animal room unless maintained in
isolator caging, racks or cabinets. Shipments of the same species, acquired from different
suppliers, should also be separated according to health status if space permits, or housed
in isolator caging. Where the mixing of species and/or stocks from different sources is
unavoidable, every effort should be made to place together those that are behaviourally
compatible, have similar environmental requirements, and a low probability of cross-
infection. NHP should not be housed with any other species.

Animals should be held in enclosures adequate for that species as described in the
individual sections of Volume 2 of this Guide.

4. Identification and Records

Cage or group identification may be used for small laboratory animals if individual
identification is not an experimental prerequisite. Individual identification can be by ear
tagging, ear notching, tattooing, tail marking, subcutaneous microchip implant or other
species appropriate method (Hayden, 1974; Ball, Argentieri, Krause et al. 1991; Iwaki,
Matsuo and Kast, 1989; Castor and Zaldivar, 1973). Dye marking on the hair provides for
short-term identification. Larger laboratory animals should always be individually
identified by tattoo, neck band, individual tag, or a subcutaneous identity tag.

The Canadian Council on Animal Care (CCAC) opposes the use of toe clipping as a
method of identification for the short-term learning experience of field studies. Where it
is necessary to provide permanent individual identification between litter members of

92
newborn rodents, toe clipping may be necessary. If, for any reason, this procedure has to
be undertaken on other than neonatal animals, either a local or general anesthetic should
be administered (Stonehouse, 1978).

The importance of keeping complete and thorough records on all experimental animals
cannot be overemphasized. The following information should be recorded for each
animal: arrival date, sex, estimated age and weight, breed and type, colour and
markings and any physical abnormalities or other identifying features (ILAR, 1984).
The name of the project or investigator and protocol number to which it is allocated
should be noted, as well as that of the supplier and eventual disposition. Animal
records should be kept for a period of one year after the final disposition of the animal.
The cages in which animals are housed, prior to or during an experiment, should be
clearly marked indicating the sex and number of contained animals, the investigator
responsible for them and such special instructions as may be pertinent to their care.
Records, especially when used in conjunction with data processing equipment, can
facilitate facility management (Wasserman, Blumrick and Liddell, 1982; Rieger and
Beriault, 1983).

Use of room cards/boards on the doors of animal rooms indicating the species,
investigator(s) whose animals are being held and any special notations that may be
pertinent is a good practice.

People donating animals to research facilities are required to sign a statement that they
are the legal owner. This document should include identification of the animal by the
criteria previously noted, and should specifically transfer ownership and disposal of the
animal to the institution. Animals (such as dogs, for which a system of national registry
exists) should always be checked for the presence of identifying markings.

C. CARE OF THE ANIMAL

1. Food

All experimental animals should receive palatable, wholesome and nutritionally adequate
food according to the requirements of the species, unless the study requires otherwise. In
certain experiments where small quantities of chemical residues may influence results,
certified diets with documented analysis of contaminant pesticides, herbicides, etc., are
available from commercial manufacturers of laboratory animal diets.

a) Food Storage

Whenever possible, pasteurized or sterilized diets obtained from reputable suppliers

should be used. Proper storage of foods is necessary to minimize the possibility of
contamination, deterioration or spoilage. Dry laboratory animal diets should be used
within six months of the milling date when stored in cool, dry, well-ventilated quarters.
Irradiated diets kept under the same conditions have approximately double the shelf life.

93
Primate and guinea pig diets should be used within three months of the milling date,
unless vitamin C is supplemented. To avoid problems from age deterioration, the date of
milling of each shipment should be obtained from the supplier (this is usually marked in
code on the bags). Bags should then be marked, put on plastic or metal pallets or racks to
keep them off the floor, and stored so that the oldest will be used first. Stale shipments
should not be accepted. Shelf life will be appreciably enhanced if the storage area is
maintained at a temperature of <16C (60.8F) (Weihe, 1987). Canned foods can be safely
stored for long periods. Clean green vegetables suitable for human consumption may
enhance the diet; however, vegetable discards may prove sources of infection and should
be avoided.

Food used in microbe-controlled environments is often autoclaved. Autoclaving

decreases the concentrations of some vitamins and antioxidants (Maerki, Rossbach and
Leuenberger, 1989). However, autoclavable diets are available which contain higher
concentrations of heat-labile ingredients to compensate for the losses induced by heat-
sterilization. Shelf life may be decreased, but need not be if the process is handled
properly (Oller, Greenman and Suber, 1985). Gamma irradiation is also used for diet
sterilization (Halls and Tallentire, 1978).

Diet in large quantities should not be stored in animal holding rooms. Small quantities,
sufficient for one or two days may be kept in the room in covered, vermin-proof
containers.

b) Special Considerations

All animals tend to reduce their food intake when sick. Animals with a high metabolic
rate, e.g., small rodents and those requiring fairly frequent feedings of high protein diets
(e.g., the cat), can become debilitated very rapidly. In cases of anorexia in these species,
oral intubation and force feeding as well as intravenous therapy (cat) should be instituted
without undue delay. Restricted feeding for maintenance of adult animals is commonly
practised for some species and strains, such as rabbits. Animals on restricted food or fluid
intake for experimental purposes should be closely monitored for weight loss, signs of
dehydration, signs of stress and deterioration in health (McIntosh and Staley, 1989). It
should be noted that food and water restriction may have a marked effect on the response
of animals to toxic substances and other experimental variables (Damon, Eidson, Hobbs
et al. 1986). For some species, particularly NHP, providing a variety of foods can be
useful as a form of environmental enrichment.

Generally, food should not be scattered over the bottom of the cage, where it may be
contaminated or wasted. Exceptions to this include provision of food to newly hatched
birds and abnormal (handicapped) animals, such as mice with muscular dystrophy.

2. Water

Drinking water should be available to animals at all times, unless contra-indicated by the
experimental protocol. Tap water, even if from municipal water systems, is not sterile and

94
quickly becomes contaminated with even more bacteria after the bottle is placed on the
cage (Tober-Meyer and Bieniek, 1981). Monitoring water quality is an important aspect
of any research program, as water contamination and chemical composition can affect the
health of animals and the results of animal experiments.

Methods available to remove both microbial and chemical contamination include

acidification, chlorination, reverse osmosis, ultrafiltration and ultraviolet (UV) light
(Newell, 1980). Some of these methods may alter immune function (Herman, White and
Lang, 1982; Fidler, 1977) and growth rates in experimental animals (Hall, White and
Lang, 1980; Tober-Meyer, Bieniek and Kupke, 1981). Regardless of whether or not the
water supply is treated, all water dispensing equipment should be thoroughly sanitized
according to institutional SOPs, and periodically monitored for bacterial contaminants.

A watering method unlikely to spread disease or contaminate the water supply should be
chosen. Water bottles should be transparent so as to permit ready observation of
cleanliness and water level; of a material that will withstand sterilization, and of a wide
mouth design to facilitate cleaning. Water bottles should always be replaced with clean,
freshly filled ones, rather than by refilling the ones in use. Animals housed under freezing
conditions may require heated water bowls.

Automatic watering devices are economical to operate, but if not properly designed, are
difficult to disinfect properly and may lead to cross-contamination (Malatesta and
Schwartz, 1985). Recirculating systems eliminate stagnation of water and help prevent
buildup of microorganisms. The correct pressure in the drinking valves prevents
backflow of water into the lines when animals drink from or play with the valve.
Malfunction of automatic watering systems can lead to drowning or drought;
consequently, the system must be routinely and thoroughly checked. Some animals need
to be taught to use automatic watering devices. Automatic watering devices are not
recommended for guinea pigs, unless they are habituated to them.

Most fish have a low tolerance for both copper ions and chlorine. Their water supply,
therefore, should either be dechlorinated or obtained from an untreated source, and
should not be brought into the aquarium through copper piping.

3. Exercise

Experts disagree about the need for exercise in laboratory animals. A judgement in such
cases must thus be made by the laboratory animal veterinarian in consultation with the
investigator. Although many adult animals do not seem to have a motivation to exercise
per se, in the process of satisfying their behavioural needs, they do get exercise (Fox,
1990). Exercise requirements for animals should reflect species, age and environment.
Research information on the requirements of each species for exercise is limited and
varied, but continually increasing. Young animals of most species involve themselves in
much more play and exercise activity than adults. For some species, exercise may not be
required in adult animals for physiological health (Weihe, 1987; Clark, 1990; Campbell,
1990). Several studies suggest that there are no beneficial effects on behaviour, health or

95
in enhancement of voluntary activity in the laboratory-bred beagle from increasing the
cage dimensions beyond the standard 76 cm x 76 cm x 76 cm (30" x 30" x 30") size,
provision of half-hour daily exercise, or from 1.22 m x 3.05 m (4' x 10') floor pen
housing (Newton, 1972; Hite, Hanson, Bohidar et al. 1977). Judgment should be based
on the animal's breed, temperament, physical condition, the conditions under which it has
previously been kept and the length of time it is to be confined. Animal cages must,
however, always be large enough to allow the innate normal behavioural and postural
adjustments (see Appendix I). There are many varied methods and programs of exercise
which are successfully used in dogs (Eckstein, Moran, Gomez et al. 1987; Clark, 1990;
Hughes and Campbell, 1990), including walking programs using outside volunteers.
Caged rats spontaneously exercise by playing with cage mates and during feeding
(Weihe, 1987) (see also Social and Behavioural Requirements of Experimental Animals).

D. CARE OF THE FACILITY

1. Cleaning and Sanitation

Employees must be aware of proper cleaning and disinfecting procedures and their
importance in disease prevention (Small, 1984; Harrison and Mahnke, 1991; Van Houton
and Hayre, 1991). All cages, pens, racks, aquaria, accessory equipment, etc., must be
thoroughly cleaned and disinfected before reuse. Most of these items should be subject to
regular (usually weekly) cleaning during use. As a general rule, laboratory animals
should be moved to freshly cleaned cages at least once a week. Cleaning practices
need to be modified according to the species and housing system for domestic animals,
fowl, reptiles and aquatic animals. The effectiveness of detergents, disinfectants and
facility cleaning programs should be monitored and constant (Thibert, 1980).

The ability to clean and sanitize a facility is greatly influenced by facility design and
construction materials. The objective of a sanitation program is to reduce the microbial
contamination or "bioburden" to a level that reduces the possibility of any cross-
contamination (Harrison and Mahnke, 1991). Proper sanitation will not compensate for
the transfer of infection by personnel. Cleaning and sanitation merely complement proper
procedures which minimize contamination (Thibert, 1980). Activities such as pressure
spraying and dumping bedding can aerosolize microorganisms allowing cross-
contamination if animals are present (Frost and Hamm Jr., 1990). Opening doors can alter
the airflow in a facility, enhancing the possibility of transfer of contaminants (Keene and
Sansone, 1984). Moveable equipment can transmit organisms between areas. Therefore,
such equipment should be dedicated to a particular room or area.

Procedure rooms using animals from different sources are a potential source of cross-
contamination. Proper disinfection of surfaces should be ensured after use.

Bedding in animal cages or pens should be changed as often as necessary to keep the
animal clean, dry, and relatively odour-free and ammonia levels in the cage at appropriate
levels. In rats, this is 25 ppm (Schoeb, Davidson and Lindsey, 1982). Smaller laboratory

96
animals require one to three changes per week, depending on such variables as the sizes
of the animals, population density and type of caging and whether or not litters are being
produced. Larger species such as dogs, cats and NHP usually require at least a daily
change.

Food containers should be easily cleaned and disinfected.

Animal cages are most efficiently cleaned and sanitized with mechanical washing
equipment operating at 83C (180F) or higher, for a minimum of ten minutes. Cages
should be carefully rinsed to remove all traces of washing and disinfecting agents, as
exposure to these may adversely affect both the animal and the experimental results. All
automatic washing equipment should be subjected to regular maintenance to assure
proper performance. Where an automatic cagewasher is not available, use of a spray
washer and disinfectant are preferable to the dip tank and rinsing method. It should be
noted that sodium hypochlorite and iodophores are effective on most animal viruses;
however, disinfectants should be chosen according to the spectrum of viruses and
organisms required to be killed and the possibility of deactivation by the local
environment. There are references available to aid in identification of the appropriate
disinfectants (Block, 1983; Harrison and Mahnke, 1991; Orcutt, 1991). Chlorine dioxide
sterilants/disinfectants have become more recently available and are often used in
facilities maintaining SPF or immunosuppressed animals because of their rapid broad
spectrum activity, even in the presence of an organic load (Frost and Hamm Jr., 1990).

All chemicals should be used properly, according to label directions. Detergents,

disinfectants and pesticides may cause changes in the experimental animal by inducing or
inhibiting cellular enzyme activity (Burek and Schwetz, 1980). This should be a
consideration when conducting experiments which may be adversely affected.

2. Waste Disposal

Dead animals, animal tissues and excreta, bedding, unused food, etc., should be collected
in leak-proof metal or plastic containers with leak-proof, disposable liners and tight lids.
Liners are essential for animal tissues, carcasses, and radioactive or toxic waste.
Infectious waste should, ideally, be incinerated on the site. If the waste is to leave the
facility it should be sterilized (autoclaved) before removal. Gamma irradiation is a
relatively recent method of disinfection of waste products which may come into more
prominent use (Garcia, Brooks, Stewart et al. 1987).

Waste which cannot be rapidly disposed of should be stored in a cold storage area
provided for that purpose. Such areas must be vermin-free, easily cleaned and disinfected
as well as being physically separated from other storage facilities. The waste storage area
should be located so that wastes need not be carried through other rooms of the facility.

Dead animals should be removed from cages as soon as they are noticed. The laboratory
animal veterinarian who should have been immediately informed of sick animals, should
also be informed of dead ones. Dead animals should be properly identified, placed in

97
disposable plastic bags and taken to the postmortem area immediately upon discovery. In
the postmortem area they should be held under refrigeration for necropsy or for disposal
in accordance with the investigator's instructions. National guidelines as well as local and
provincial laws control waste disposal practices that could endanger public health
(HWC/MRC, 1990). Saskatchewan, Alberta and New Brunswick regulate livestock
management and manure disposal, and Ontario has a suggested Code of Practice on the
same subject. (Copies of these may be obtained from provincial departments of
Agriculture.)

Considerable forethought and extensive consultation is advisable before installing an

incineration facility for the disposal of pathological waste.

3. Vermin Control

A properly constructed building should be vermin-proof, but may not be free from
vermin. Vermin enter on food, bedding, people and animals. Insects and arthropods thus
introduced, may act as the intermediate hosts of certain parasites and may also
mechanically transmit bacterial and other pathogens (Hughes, Kassim, Gregory et al.
1989). Wild rodents may transmit a wide variety of bacteria, viruses, and parasites to
caged members of closely related species (Levine and Lage, 1984). New facilities should
be checked critically for vermin before any animals are moved in.

Vermin should also be controlled in already-infested older buildings. A control program

will include the proper training of personnel, good waste disposal, sealing or eliminating
breeding sites, extermination through pesticides or trapping, and the recovery of all
escaped and/or wild animals. It is important that pesticides be applied only under
professional supervision. Many pesticides are dangerous to humans, and may adversely
affect the experimental animal and the research (Bell, Farrell and Padgett, 1975). Any
control program that is initiated must extend throughout all areas of the facility, with
special attention to food and bedding storage. The practice of using a free-roaming cat for
the control of wild and escaped rodents is not acceptable except in farm animal facilities,
and only under close management.

If insect colonies are kept in or near an animal care facility, there must be regular
monitoring of the facility against infestation from escapees. Such insect colonies should
be kept behind a screened enclosure or inside an escape-proof container. The use of
insecticides must also be compatible with these insect colonies.

4. Holiday and Emergency Care

a) Weekend and Holiday Care of Laboratory Animals is Essential

It should be recognized that changes in personnel and feeding and cleaning schedules, as
can occur during these periods, are known to be stressful to routine-oriented animals
(Beaver, 1981).

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b) Animal Care is a Continuous and Daily Responsibility

This point should be emphasized in job descriptions for animal care personnel and in
union contracts. Basic animal care should be categorized as an "essential service" and a
clause to this effect should be included in all collective agreements, and should not be
subject to interruption through strike action. Staff must be provided for weekends and
holidays, and skilled assistance must be available in the event of an emergency.

The names and telephone numbers of staff responsible for the animals should be given to
security personnel. Some institutions may also choose to have contact telephone numbers
posted prominently in the facility. In either case, directions for contacting responsible
animal care staff must be made available in the facility. All the animal care staff should
be informed of their responsibilities in emergency situations.

The CCAC suggests use of the following:

Essential Services Provision

To be inserted near the Strikes and Lockouts clause:

Clause

"Designation of Employees to Care for Research Animals

The parties agree that proper care* of all research animals** will be maintained by the
members of the bargaining unit in the event of a strike or lockout in the course of this
Agreement or its continuance.

At least seven days before the commencement of a strike or lockout, the employer will
designate and identify a number of employees which it deems sufficient to provide for
continuous proper care of the animals during the strike or lockout. A list of the names will
be delivered to the Union and the parties agree to meet with a view to executing a formal
agreement with respect to the employees affected. Should the parties be unable to reach
agreement on the persons to be designated, the matter will be referred to the CCAC, for
final and binding resolution by the Council.

All persons so designated will be paid their regular salary during the period of
designation.

Due regard will be had for previously arranged vacations and other matters and as far as
possible the designated duties will be dispersed among all appropriate employees equally.
No other duties will be assigned to these designated employees.
____________________

* Proper care implies provision of appropriate temperatures, humidity, light cycles,

ventilation, food, water and cleaning as well as

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 exercise and nursing care where appropriate.

** Research animals means any live non-human vertebrate or invertebrate utilized in

research, teaching and testing."

E. REFERENCES

AGRICULTURE CANADA. Publication 1771/E. Recommended code of practice for

care and handling of pigs. Communications Branch, Agriculture Canada, Ottawa, Ont.,
K1A 0C7. 1984.

IBID. Publication 1898/E. Recommended code of practice for the care and handling of
farm animals--pigs. In Press.

IBID. Publication 1821/E. Recommended code of practice for the care and handling of
special fed veal calves. 1988.

IBID. Publication 1757/E. Recommended code of practice for the care and handling of
poultry from hatchery to processing plant. 1989.

IBID. Publication 1853/E. Recommended code of practice for the care and handling of
dairy cattle. 1990.

IBID. Publication 1870/E. Recommended code of practice for the care and handling of
farm animals--beef cattle. 1991.

BALL, D.J., ARGENTIERI, G., KRAUSE, R. et al. Evaluation of a microchip implant

system used for animal identification in rats. Lab. Anim. Sci. 1991; 41(2): 185-186.

BEAVER, B.V. Behavioural considerations for laboratory dogs and cats. Compend. Cont.
Ed. 1981; 2: 212-215.

BELL, T.G., FARRELL, R.K. and PADGETT, G.A. Ataxia, depression and dermatitis
associated with the use of Dichlorvos impregnated collars in the laboratory cat. J. Am.
Vet. Med. Assoc. 1975; 167: 579-586.

BLOCK, S.S., ed. Disinfection, sterilization and preservation. 2nd Ed. Philadelphia, PA:
Lea and Febiger, 1983.

BUREK, J.D. and SCHWETZ, B.A. Considerations in the selection and use of chemicals
within the animal facility. Lab. Anim. Sci. 1980; 30: 414-419.

CAMPBELL, S.A. Effects of exercise programs on serum biochemical stress indicators

in purpose-bred beagle dogs. In: Mench, J.A. and Krulisch, L., eds. Canine research
environment. Bethesda, MD: SCAW (Scientists Center for Animal Welfare), 1990: 77-80.

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CASTOR, G.B. and ZALDIVAR, R.A. Tattooing rabbits' ears for identification. Lab.
Anim. Sci. 1973; 23: 279-281.

CAUDILL, C.J. and GADDIS, S.E. A safe and efficient handling device for wild rodents.
Lab. Anim. Sci. 1973; 23: 685-686.

CLARK, J.D. Research studies in exercise and behaviour of dogs. In: Mench, J.A. and
Krulisch, L., eds. Canine research environment. Bethesda, MD: SCAW (Scientists Center
for Animal Welfare), 1990: 61-64.

DAMON, E.G., EIDSON, A.F., HOBBS, C.H. and HAHN, F.F. Effect of acclimation to
caging on nephrotoxic response of rats to uranium. Lab. Anim. Sci. 1986; 36: 24-27.

ECKSTEIN, E.C., MORAN, D., GOMEZ, E., POMERANZ, M.L., BLOCK, N.L. and
KLINE, J. A method for tethering dogs in a run. Lab. Anim. Sci. 1987; 37: 234-235.

FALL, M.W. Use of red light for handling wild rats. Lab. Anim. Sci. 1974; 24: 686-687.

FIDLER, I.J. Depression of macrophages in mice drinking hyperchlorinated water.

Nature 1977; 270: 735-736.

FOX, M.W. Canine behavior. In: Mench, J.A. and Krulisch, L., eds. Canine research
environment. Bethesda, MD: SCAW (Scientists Center for Animal Welfare), 1990: 21-28.

FROST, W.W. and HAMM, T.E. Jr. Prevention and control of animal disease. In: Rollin,
B.E., ed. The experimental animal in biomedical research. Boca Raton, FL: CRC Press,
1990; 1: 133-152.

GARCIA, M.M., BROOKS, B.W., STEWART, R.B., DION, W., TRUDEL, J.R.J. and
OUWEWKERK, T. Evaluation of gamma radiation levels for reducing pathogenic
bacteria and fungi in animal sewage and laboratory effluents. Can. J. Vet. Res. 1987; 51:
285-289.

HALL, J.E., WHITE, W.J. and LANG, C.M. Acidification of drinking water: its effects
on selected biological phenomena in male mice. Lab. Anim. Sci. 1980; 30: 643-651.

HALLS, N.A. and TALLENTIRE, A. Effects of processing and gamma irradiation on the
microbiological contaminants of a laboratory animal diet. Lab. Anim. 1978; 12: 5-10.

HARRISON, S.K. and MAHNKE, C. Selection and use of disinfectants and sterilants.
AALAS (Am. Assoc. Lab. Anim. Sci.) Bull. 1991; 30(2): 10-14.

HAYDEN, P. (letters to the editor) Identification methods for rodents. Lab. Anim. Sci.

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1974; 24: 428.

HEALTH AND WELFARE CANADA/MEDICAL RESEARCH COUNCIL. Cat. No.

MR 21-1/1990/E. Laboratory biosafety guidelines. Ottawa, Ont.: Office of Biosafety,
Laboratory Centre for Disease Control, Health and Welfare Canada, 1990.

HERMAN, L.M., WHITE, W.J. and LANG, C.M. Prolonged exposure to acid, chlorine
or tetracycline in the drinking water: effects on delayed-type hypersensitivity,
hemagglutination titers and reticuloendothelial clearance rates in mice. Lab. Anim. Sci.
1982; 30: 603-608.

HITE, M., HANSON, H.M., BOHIDAR, N.R., CONTI, P.A. and MATTIS, P.A. Effects
of cage size on patterns of activity and health of beagle dogs. Lab. Anim. Sci. 1977; 27:
60-64.

HUGHES, D.E., KASSIM, O.O., GREGORY, J., STUPART, M., AUSTIN, L. and
DUFFIELD, R. Spectrum of bacterial pathogens transmitted by Pharaoh's ants. Lab.
Anim. Sci. 1989; 39: 167-168.

HUGHES, H.C. and CAMPBELL, S. Effects of primary enclosure size and human
contact. In: Mench, J.A. and Krulisch, L., eds. Canine research environment. Bethesda,
MD: SCAW (Scientists Center for Animal Welfare), 1990: 66-73.

INSTITUTE FOR LABORATORY ANIMAL RESOURCES (Committee on Laboratory

Animal Records, 1984). Washington, DC: ILAR/National Academy of Sciences, 1990.

IWAKI, S., MATSUO, A. and KAST, A. Identification of newborn rats by tattooing. Lab.
Anim. 1989; 23: 361-364.

KEENE, J.H. and SANSONE, E.B. Airborne transfer of contaminants in ventilated

spaces. Lab. Anim. Sci. 1984; 34: 453-457.

LEVINE, J.F. and LAGE, A.L. House mouse mites infesting laboratory rodents. Lab.
Anim. Sci. 1984; 34: 393-394.

MALATESTA, P.F. and SCHWARTZ, L.H. Use of a chemical tracer to evaluate water
movement through two automatic watering rack manifolds during flushing. Lab. Anim.
Sci. 1985; 35: 89-91.

MAERKI, U., ROSSBACH, W. and LEUENBERGER, J. Consistency of laboratory

animal food following incubation prior to autoclaving. Lab. Anim. 1989; 23: 319-323.

MCINTOSH, J. and STALEY, E.C. (moderators) Limitations of food and water

deprivation. In: Guttman, H.N., Mench, J.A. and Simmonds, R.C., eds. Science and
animals: addressing contemporary issues. Bethesda, MD: SCAW (Scientists Center for
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NEWELL, G.W. The quality, treatment and monitoring of water for laboratory rodents.
Lab. Anim. Sci. 1980; 30: 377-383.

NEWTON, W. An evaluation of the effects of various degrees of long-term confinement

on adult beagle dogs. Lab. Anim. Sci. 1972; 22: 860-864.

OLLER, W.L., GREENMAN, D.L. and SUBER, R. Quality changes in animal feed
resulting from extended storage. Lab. Anim. Sci. 1985; 35: 646-650.

OLSON, M.E., MORCK, D.W. and NABROTZKY, V.C.A. Manual of standard operating
procedures for animal facilities, 1992. Available from: CALAS/ACTAL National Office,
Bioscience Animal Service, M524 Biological Sciences Building, University of Alberta,
Edmonton, Alberta, T6G 2E9 CANADA.

ORCUTT, R.P. Evaluation of disinfectants and sterilants. AALAS (Am. Assoc. Lab.
Anim. Sci.) Bull. 1991; 30(2): 15-17.

RIEGER, D. and BERIAULT, R. A microcomputer based program for the storage and
retrieval of experimental animal information. Lab. Anim. Sci. 1983; 33: 390-394.

SCHOEB, T.R., DAVIDSON, M.K. and LINDSEY, J.R. Intracage ammonia promotes
growth of Mycoplasma pulmonis in the respiratory tracts of rats. Infect. Immun. 1982; 38:
212-217.

SMALL, D.J. Rodent and lagomorph health surveillance - quality assurance. In: Fox,
J.G., Cohen, B.J. and Loew, F.M., eds. Laboratory animal medicine. Toronto, Ont.:
Academic Press, 1984: 709-723.

STONEHOUSE, B., ed. Animal markings: Recognition marking of animals in research.

London and Basingstock: McMillan Press Ltd., 1978.

THIBERT, P. Control of microbial contamination in the use of laboratory rodents. Lab.

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TOBER-MEYER, B.K. and BIENIEK, H.J. Studies on the hygiene of drinking water for
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TOBER-MEYER, B.K., BIENIEK, H.J. and KUPKE, I.R. Studies on the hygiene of
drinking water for laboratory animals. 2. Clinical and biochemical studies in rats and
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117.

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VAN HOUTON, J. and HAYRE, M.D. Disinfectants and sterilants: their chemistry, use
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WASSERMAN, R.S., BLUMRICK, K. and LIDDELL, R.W. III. An automated

interactive animal colony management system. Lab. Anim. Sci. 1982; 32: 550-552.

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New York, NY: Churchill Livingstone, Inc. 1987: 309-330.

Chapter VI - Social and Behavioural Requirements of Experimental Animals

VI. SOCIAL AND BEHAVIOURAL REQUIREMENTS OF

EXPERIMENTAL ANIMALS

A. INTRODUCTION

In the past, emphasis has been directed towards providing adequate caging for
experimental animals in order to contain them hygienically, to facilitate husbandry, and
minimize (husbandry) variables. However, increasing importance is now being placed on
reducing the animal's stress, and improving its social and behavioural well-being.
Provision of varied environmental enrichment may or may not result in increased cost of
operation; however, it is considered that there is often immediate benefit to the animal
and ultimately to the researcher and the research.

This chapter contains general principles rather than specifics. It is not infallible. Nor
should it be taken literally at the expense of the animal (for example, it may be found that
an otherwise desirable environment or social grouping is not suitable for an individual
animal). This chapter, including a CCAC-approved policy statement, will receive
continuing review, with changes made as needed. The ability to treat animals in the way
we would wish requires sensitive and conscientious applications of knowledge. Critical,
rigorous, scientific endeavour is required of us all in order to reach this goal.
Management and housing situations that fulfil the animal's behavioural requirements
should be interpreted as providing an ideal toward which we should aim.

1. What is Animal Well-being or Welfare?

Animal welfare is described by Broom (1986) as an animal's "state as regards its attempts
to cope with its environment". Blood and Studdert (1988) define it as "maintaining
appropriate standards of accommodation, feeding and general care, the prevention and
treatment of disease..." The American Veterinary Medical Association (AVMA) enlarges
on this to include "all aspects of animal well-being, including proper housing,
management, nutrition, disease prevention and treatment, responsible care, humane

104
handling, and, when necessary, humane euthanasia" (Anon., 1990).

Fraser (1989) notes that animal well-being encompasses "both the physical and
psychological. These normally coexist. Physical well-being is manifested by a state of
clinical health. Psychological well-being is reflected, in turn, in behavioural well-being.
The latter is evident in the presence of normal behaviour and the absence of substantially
abnormal behaviour."

The World Veterinary Association (WVA) states that animal ethology "puts the emphasis
on knowledge which is scientifically based. Its aim is to clarify: a) needs that can be
filled; and b) harm that can be avoided..." (WVA, 1989).

Hurnik (1988) defines animal well-being as "a state or condition of physical and
psychological harmony between the organism and its surroundings." However, it is
agreed that animal welfare is not a single phenomenon, and that on one definition will
satisfy everyone (Moberg, 1992; Baxter, 1993; Duncan and Dawkins, 1983).

The Royal Society for the Prevention of Cruelty to Animals (RSPCA) has recently
recognized the need to draw attention to stress in experimental animals and the necessity
for alleviating this condition whenever it is associated with suffering (RSPCA, 1992). An
annotated bibliography on animal welfare has recently been prepared (Murphy, Rowan
and Smeby, 1991).

One must always remember, as well, Hollands's sensitive definition written more than a
decade ago: "This then would be my definition of animal welfare: dignity-according to
animals the natural dignity which is due them as living, sentient creatures" (Hollands,
1980).

2. Environmental Enrichment

Environmental enrichment is defined by Beaver (1989) as "additions to an animal's

environment with which it can interact."

As a general rule, most experimental animals are social animals and benefit from the
company of conspecifics or humans. As well, predictability in interactions usually
enhances the animal's well-being, while the opposite results from frequent regrouping
and restabilizing.

It should be remembered that an animal's experiences during the developmental phases

determine social behaviour. Therefore, conditions of animal holding in a breeding facility
will impact on the animal's future well-being.

The social needs of animals used in research, teaching, or testing, should be given equal
consideration with environmental factors such as lighting, heating, ventilation and
containment (caging). Particularly in the case of singly housed animals, daily observation
provides an alternative form of social contact for the animal and commonly facilitates

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handling in that the animal becomes accustomed to the human presence.

A more complex environment, use of artificial appliances, and better use of existing
space enhance stimulation. Simply increasing the number of square inches or centimetres
available to an animal does not promote better space utilization (Line, 1987; Fajzi,
Reinhardt and Smith, 1989); however, the amount of space should be appropriate to the
species. In group housed animals, the size of the social group in relation to its available
space should be regularly appraised.

3. Group Formation

When animals are introduced to each other and pairs or groups are established, there is an
initial period during which they work out their social relationships (dominance ranks,
etc.). There may be aggressive interactions; however, when conditions are right, the
social organization will stabilize. Once the hierarchy has been established the interactions
are subtle, and based more on avoidance or ritualized threat than overt aggressive action.
If their daily routine is disrupted, if resources such as food or resting spaces are limited,
or the animals are poorly grouped, the hierarchy becomes disestablished and the number
of aggressive interactions increases. The animal's well-being is threatened when:

a) space is insufficient for maintaining behaviourally adequate distance;

b) feeding or resting space for all individuals is insufficient; or when feeding and resting
cannot be accomplished concurrently;

c) regrouping is performed so frequently that animals must repeatedly undergo the

stabilization process; and

d) group sizes are inappropriate for the species.

The above statement challenges intense confinement practices which prohibit animals
from engaging in their normal social-behavioural activities.

In addition to sufficient primary space for resting, animals also need what could be called
secondary space, for freedom of movement at their own will. An important exception
may occur at the time of parturition, when most individual animals should be given their
own quarters.

Most animals should not be housed singly unless required by medical condition,
aggression, or the dictates of the study. Singly housed animals should have some degree
of social contact with others of their own kind. For most species, at the very least there
should be potential visual contact. Olfactory and auditory contact with other animals is
also usually desirable.

Protocols which involve single housing must describe proposed measures for
meeting the social requirements of the isolated animal (e.g., where appropriate,

106
increased positive human contact). Investigators must justify any deviations from
the CCAC guidelines before an Animal Care Committee (ACC) and receive its
approval, before any study can begin. All protocols must be reviewed at least
annually by the ACC.

4. Position Statement

"SOCIAL AND BEHAVIOURAL REQUIREMENTS OF

EXPERIMENTAL ANIMALS (SBREA)

Well-being in animals has two components: physical and behavioural. Physical well-
being is manifested by a state of clinical health. Behavioural well-being is manifested by
behaviour considered to be normal for that species and strain, together with the absence
of significantly abnormal behaviour. Behavioural well-being is considered to reflect
psychological well-being, and to that extent, the terms are considered to be synonymous
in our usage.

In the interest of well-being, a social environment is desired for each animal which will
allow basic social contacts and positive social relationships. Social behaviour assists
animals to cope with circumstances of confinement. Caging, whether for single animals,
pairs, or groups, should be enriched appropriately for the species.

It is necessary to recognize affiliations which commonly occur within and between

species. Chronic isolation as a method of accommodation, should not normally occur.
However, in exceptional circumstances, and with clear scientific and biological
justification, some animals may be better kept alone. Positive interactions with human-
beings are important in some species, and particularly in conditions of social isolation.
Some individuals seem more readily accepted by animals than others; this concept should
be used to maximize the benefits of these affiliations.

February, 1990
B. ANIMALS USED IN AGRICULTURAL RESEARCH

1. Introduction

The report of the Brambell Commission in 1965, described the farm animal's "Five
Freedoms" as the ability to easily "turn around, groom itself, get up, lie down, and stretch
its limbs" (Brambell, 1965). In 1989, the WVA adopted its own Five Freedoms, which
applied to all species and which were based on those of Britain's Farm Animal Welfare
Council (FAWC) (Webster, 1987). The FAWC has recently revised these Five Freedoms,
which define ideal states and which now include:

a) Freedom from hunger and thirst;

b) Freedom from discomfort;
c) Freedom from pain, injury and disease;

107
d) Freedom to express normal behaviour;
e) Freedom from fear and distress (Seamer, 1993).

Carpenter (1980) suggested enough freedom to perform natural physical movement,

including daily routines of natural activities; facilities for comfort activities such as rest,
sleep, and body care; adequate food and water to maintain full health; social contact with
other animals of their own kind; opportunity for exploration and play, especially in young
animals; and satisfaction of minimal spatial and territorial requirements.

The necessary scientific knowledge upon which to base a comprehensive set of

guidelines which would fully protect the welfare of agricultural animals is not yet
available (Expert Committee on Farm Animal Welfare and Behaviour, 1987).
Nonetheless, the CCAC believes that there is sufficient information to set out some
general principles that can be updated and expanded as

Codes of Practice for Canada for pigs, veal calves, poultry, dairy cattle and beef cattle
(Agriculture Canada, 1771/E, 1984; 1821/E, 1988; 1757/E, 1989; 1853/E, 1990; 1870/E,
1992) represent the basic industry standards and, for the most part, are the minimum
CCAC requires for research institutions undertaking agricultural research. In addition, a
revision of the Recommended Code of Practice for the Care and Handling of Farm
Animals--Pigs (1898/E) is now in press. Researchers and others working with agricultural
animals must be fully conversant with these codes.

The Health of Animals Act (Government of Canada, 1990), which replaced the Animal
Disease and Protection Act, states that the "Governor-in-Council may make regulations:

a) for the humane treatment of animals and generally:

i) governing the care, handling and disposition of animals;

ii) governing the manner in which animals are transported within, into or out of Canada;
and

iii) providing for the treatment or disposal of animals that are not cared for, handled or
transported in a humane manner."

Curtis (1992) suggests that there is a double standard in place regarding current farm
practices and the procedures used with agricultural animals in biomedical research. This
is a dilemma not uncommonly faced by ACCs. Another dilemma occurs because
agricultural animals are used in food and fibre-related research, as well as biomedical
research, and it is often difficult to clearly label such studies as being only agricultural or
only biomedical (Stricklin, Purcell and Mench, 1992).

In attempting to develop guidelines, thought, observation, and concern for the animal
must be uppermost in our minds for, as Hurnik (1988) states: "There is a need to exercise
extreme care to avoid the emotional tendency to rely exclusively on characteristics which

108
may be of concern to humans, but that are not necessarily central to the overall quality of
animal life." Spedding (1988) also warns that "perhaps the biggest danger is that a
dissatisfied public may demand changes that eliminate what is disliked, but result in
either no improvement or even a reduction in the welfare of the animals involved."

In that we must "work to ensure the animal's welfare in life and at the point of death"
(Webster, 1987), we might note a recent example of change resulting from human
perception of what constitutes humaneness, when Great Britain passed Slaughter of
Animals (Humane Conditions) Regulations 1990 (effective July 5, 1992), requiring head
restraint during slaughter of cattle. Unfortunately, after the fact, research showed, by
measuring cortisol levels, that the restraint process was far more stressful than allowing
the animal to stand free when the captive bolt was fired (Ewbank, Parker and Mason,
1992).

2. Animal Stress

Suffering, loosely defined as experiencing a wide range of unpleasant emotional states

such as pain, fear, anxiety, frustration and perhaps boredom, can be a major threat to an
animal's welfare. Poor animal welfare may be apparent in changes in an animal's
behaviour, physiology, health status, reproduction or growth. Many clinical conditions in
animals first become apparent to observers through a set of behavioural indicators
(Fraser, 1984/85). Depressed animals show a depletion of the behavioural repertoire
characteristic of the normal animal (Fraser, 1984/85, 1988).

The three ways in which an animal responds to a stressful situation include changes in
behaviour, activation of the autonomic nervous system, and activation of the
neuroendocrine system. Because of its rapid and specific responses to many stressors, the
autonomic nervous system has been an appealing candidate for the diagnosis of stress by
measuring heart rate, respiration, and the secretion of catecholamines. Many investigators
accept the increased secretion of glucocorticoids as proof of stress occurring. It has been
demonstrated that stress associated with transportation, restraint or handling diminishes
immune function in a number of livestock species (Grandin, 1992; Kelley, Osborne,
Evermann et al. 1981; Coppinger, Minton, Reddy et al. 1990).

However, Moberg (1985) states that monitoring these autonomic nervous system and
endocrine system responses outside the laboratory has not been practical, and thus they
have proven to be of little use in defining stress and well-being in domestic animals.
Duncan (1992) notes that sophisticated biological systems have evolved to help animals
cope with stress, and that, while it is impossible for us to shield agricultural animals from
all stress, the key to protecting their well-being is to minimize the biological costs of
unavoidable stress, and to recognize the need for research directed at stress.

The idea that animals have certain "behavioural needs" has received considerable
attention. For example, Baxter (1983) has argued that all psychological processes which
affect animal welfare must have a desired state or set point (or set band covering a range
of values) at which the animal will strive to remain, or to which it wishes to return.

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Deviations from this set point will cause a reduction in the animal's well-being. However,
Hughes and Duncan (1988) reviewed the literature in this area and showed that, in certain
cases, an argument could be made for animals needing to be able to perform particular
behaviours, even if the goal of the behaviour has already been provided to the animal.

A behavioural need will appear in the cases of behaviour motivated mainly by internal
factors or by a complex interaction between internal and external factors where the
performance of the pattern is constrained. For example, a hen provided with a replica of a
nest is still motivated to perform normal nest building behaviours (Hughes, Duncan and
Brown, 1989).

3. Housing and Husbandry

Three major factors influence welfare in animal agriculture: a) housing; b) stockmanship;

and, c) management (Hurnik, 1988). Hughes and Duncan (1988) suggest that, in order to
fully protect welfare, a housing system would have to allow the performance of certain
behaviour patterns in addition to providing all the environmental needs of the animals.
However, which behaviour patterns are essential has yet to be elucidated, and research is
being undertaken to answer this question (e.g., Dawkins, 1990).

In the U.S., the American Association for the Accreditation of Laboratory Animal Care
(AAALAC)(1991), takes the position that, in accredited facilities, housing and care for
farm animals should meet the standards that prevail on a high quality, well-managed
farm.

In maintaining farm animals in the laboratory, it is important to be aware of their

behaviour in a farm situation. A prominent feature in farm animal behaviour is the active
way in which individuals associate with each other and form social groups (Mench,
Stricklin and Purcell, 1992; Fraser and Broom, 1990; Grafen, 1990). Isolation from
conspecifics is a profound source of stress (Grandin, 1992; Gross and Siegel, 1981), as
social animals obtain both physical and physiologic comfort from each other (Friend and
Dellmeier, 1988; Van Putten, 1988).

Proper handling and restraint of farm animals in a laboratory environment are key
elements to their success as research animals. Prior familiarizing of experimental animals
to handlers and experimental procedures can be of considerable benefit in the laboratory,
says Panepinto (1992), the developer of the Panepinto sling used to restrain miniature
swine and sheep. Stroking the pig, as opposed to stressing it, has been shown to have a
beneficial effect on reproduction (Panepinto, 1992; Anon., 1992).

Animals raised in a barren, non-stimulating environment may display stereotypies such as

bar biting, cribbing or tongue rolling in horses (Van Putten, 1988; Fraser and Broom,
1990; Fraser, 1992). Environmental enrichment will often reduce unwanted excitability
and help prevent abnormal behaviour (Grandin, 1992). For example, in housing domestic
poultry used in experimentation, nest boxes and perches (high and low) should be
provided. Aquatic fowl should have access to a swimming facility or some form of

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wetting in clean water.

Animals should be housed singly only when the experimental procedure requires this;
e.g., in metabolism studies, some infectious disease studies or nutritional research.

When animals are introduced to each other and a group is established, there is an initial
period during which the animals often fight in an effort to work out their social
relationships (McGlone and Curtis, 1985; Fraser and Rushen, 1987; Fraser and Broom,
1990; Mench, Stricklin and Purcell, 1992). Subsequently, dominance-subordinate
relationships continue; however, interactions are subtle, based more on avoidance or
ritualized threat. Animals should be regrouped as infrequently as possible, so that they
need not repeatedly endure the stabilization process (Fraser and Broom, 1990; Kenny and
Tarrant, 1982).

In housing domestic species, Mench, Stricklin and Purcell (1992) suggest that there
should be enough space for maintaining some minimum separation from each other,
equal access to feed and water, and the ability to engage in significant behaviours and
make normal postural adjustments. However, it has been demonstrated that when animals
are group housed, the area available to a particular animal does not consist just of its
individual space, but instead the entire area in which the animal is enclosed. Therefore,
the space requirements per animal are greater when one houses one or few individuals
(Stricklin, Purcell and Mench, 1992).

The animal should also receive, on a regular and substantial basis, attention from an
attendant trained to deal with the species (Kilgour and Dalton, 1984). It is noted that the
domestication of various species has depended on their capacity for social affinity with
humans (Gross and Siegel, 1982; Gonyou, 1991). Substantial periods of observation must
be maintained by the personnel responsible for the well-being of these animals (e.g.,
animal attendants, veterinarians) and adequate provision made in the work-time schedule
to permit regular appraisal of the balance between social group and space.

A stockperson's quality will depend on his or her ethical sensitivity, familiarity with the
animals, skill in interpreting behavioural symptoms indicating deprivation, suffering and
morbidity, and the care shown in handling animals. The stockperson's skill in carrying out
particular tasks such as castration, injecting animals, clipping teeth, etc., is also very
important. Objective studies are now being carried out to identify the characteristics of
good stockmen (Seabrook, 1984, 1987; Hemsworth, Barnett, Coleman et al. 1989). The
quality of management refers to such things as decisions about operation of ventilation
systems, provision of food and water, provision for emergencies, provision of sanitation
and prophylactic measures, and choice of techniques and procedures for castration,
dehorning, giving injections, etc.

The quality of a housing and husbandry system can affect welfare in many different
ways. It can, of course, act in a direct physical way by causing injury, by reducing health,
or by providing climatic conditions which are far from optimal. It may also reduce
welfare by affecting the behaviour, or the physiological and immune systems of the

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animals.

Duncan (1981, 1983) has proposed the following classification for physical and social
effects of husbandry systems.

a) Physical Effects

Aspects of the physical environment provided may reduce welfare by altering the
animal's behaviour by: 1) blocking or frustrating the performance of a particular activity;
2) failing to provide the specific releasing stimuli necessary for eliciting certain
behavioural patterns; and 3) providing too high a level of general stimulation. For
example, if the environment is too complex or keeps changing in a way that the animals
cannot predict, the animals may then become fearful and anxious. Alternatively, if the
environment is too barren and monotonous, the level of general stimulation may be too
low, leading to boredom. While it is not easy to measure fear and boredom in a scientific
way, we must not assume that animals do not experience these emotions.

b) Social Effects

Husbandry systems can also influence the behaviour and welfare of animals by altering
and controlling the animals' social environment. All of the common agricultural species
are gregarious, which means that an inadequate social environment can be expected to
reduce welfare. Compared to what might be considered as "normal" or "natural," many
husbandry systems often deviate in the following ways: 1) the parent-offspring bond may
be disrupted or prevented from forming; 2) young animals may be weaned too early; 3)
animals may be kept in groups that are too large or small; 4) animals may be kept at too
high a density; 5) animals may be kept in single-age or single-sex groups; 6) group
membership may be disrupted; and 7) animals may be isolated to some extent.

4. General Principles

In order to improve and enhance the animal's environment, the CCAC encourages
research institutions:

a) to experiment with group housing for such animals as lactating sows (after a few days
in farrowing crates), calves, dairy cows and sheep. It is recognized that group housing
may lead to increased aggression or bullying among animals, increased chance of disease
transmission, and increased difficulty in detecting health problems of individual animals.
However, unless the welfare or safety of an animal is in danger, these facts should not be
used to reject group housing. The level of animal management will probably have to
increase and the training required for stockpersons will have to change. Evidence is
accumulating that new piggeries should not have individual stalls for gestating gilts or
sows unless required for experimental purposes (Barnett, Winfield, Cronin et al. 1985;
Barnett and Hemsworth, 1991; Becker, Ford, Christenson et al. 1985; Cronin, Van
Tartwijk, Van Der Hel et al. 1986; Schouten, Rushen and De Passill, 1991; Von Borell
and Ladewig, 1989).

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b) to practise environmental enrichment such as providing toys for pigs, "teats" for veal
calves, small orifice nipples and frequent feedings for artificially raised lambs, and
greater opportunities for animals to perform normal food searching and foraging
behaviours.

c) to provide the means for increased social contact and to allow the animals to perform a
wider range of behaviours.

d) to increase the age at weaning. For example, piglets weaned at three weeks have an
increased incidence of belly nosing (an abnormal behaviour); the recommended
minimum age for weaning is therefore four weeks.

e) to shorten periods of isolation and restraint and to use them only when it is absolutely
necessary, and not merely for the convenience of the experimenter. Individually penned
animals should be allowed to maintain visual contact with at least one other animal while
standing or lying in the pen, unless the isolation is required for experimental purposes
and has been approved by the ACC. For pigs, in particular, maintaining olfactory contact
may be as important as visual contact. In sheep, the head region is the primary focal point
used to recognize each other and should therefore be the least obstructed region from the
perspective of neighbouring sheep trying to maintain visual contact.

The Expert Committee on Farm Animal Welfare and Behaviour (1987) suggests that
government agencies and universities should re-order their priorities so that research on
farm animal welfare and behaviour will have a level of staffing and support typical of that
supplied to other disciplines such as nutrition, reproductive physiology, genetics, and
food products.

The CCAC recognizes the importance of education in improving animal well-being and
welfare. Ideally, all students of animal production and veterinary medicine should receive
instruction in farm animal behaviour, animal welfare, and the ethics of livestock
production.

C. ANIMALS (LARGE) HELD IN METABOLISM CAGES

The use of metabolism cages or crates necessarily reduces the animal's social and
behavioural activities. This procedure should not, therefore, be used merely for the
purpose of convenient restraint, but should be reserved for approved metabolic studies.
Animals so housed should be under close and expert observation throughout the period of
the study.

1. Conditioning

A seven to 10 day conditioning period in a floor pen, to "acclimatize" the animal to a new
diet (if this is necessary), and before its placement in the associated metabolism crate, is

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required, followed by three to four days' adjustment to the crate.

2. Size of Metabolism Crates

Enough space must be provided for the animal to rise and lie down normally. Some
animals (e.g., calves and sheep) swing their weight forward when rising; therefore, the
required length of crates should be greater than the simple length of the animal. Width of
the crates must be sufficient to allow sternal recumbency.

Other postures for example shown by sheep, as well as serving to make the animal
comfortable, also have a thermoregulatory function. If the dimensions of a metabolism
crate do not permit such postures (e.g., lateral recumbency), then proper temperature and
other environmental control becomes a responsibility of the research.

3. Contact with Other Animals

Many animals are highly social. An isolated animal is often not normal behaviourally, nor
possibly metabolically. To minimize stress, crates should be designed and positioned so
that there is good visual, auditory, and olfactory contact with conspecifics.

4. Pre-, During and Post-Experiment Checks

A physical and behavioural assessment of the animal should be done before, during, and
after an experiment. Animal care personnel should observe the animal before and after
eating in order to ascertain, for example, if intake has decreased.

5. Observing Changes in Behaviour

Strict attention should be paid to observing changes in behaviour, which can indicate a
degree of stress or anxiety, or fear stereotypies (e.g., increased drinking in sheep). Noting
such changes is important to good science as well as good animal care.

6. Duration of Confinement

Enforced immobility has a negative effect on bones, joints, and muscles. For this reason,
animals should be released for periodic exercise or released from the metabolism crates
for at least three hours per seven days.

Any period of study exceeding 21 days in a metabolism crate must be justified to the
ACC by the investigator, on grounds of experimental design and scientific merit.
However, the total period in the crate should not exceed 30 days.

7. Exceptional Circumstances

In exceptional circumstances (e.g., catheterization studies), it may not be possible to

implement the recommended weekly exercise requirement. In such cases, variances to

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these guidelines require justification by the investigator, and review and approval of the
institutional ACC.

D. CATS

1. Introduction

Various authors have proposed schemes to assist those who are attempting to enhance the
welfare of research animals. Beaver (1989) proposes five basic methods which can be
used to alter an animal's environment so as to permit the animal to live and produce to its
full potential. These include behavioural enrichment, social peers, artificial appliances,
food gathering activities, and control of the environment.

Spinelli (1989) in commenting on Beaver's five methods, disagrees with her definition of
enrichment. However, he notes that there are a variety of strategies that will be useful
singly or in combination to promote the psychological well-being of laboratory animals.
Spinelli contends that environmental enrichment and an animal's psychological well-
being "may be one of the most important areas of study in laboratory animal science over
the next few years."

Beaver's five areas will be interpreted in light of recognized behaviour systems for cats
(Felis catus). Such systems represent species-typical behaviours which are co-ordinated
to serve a specific function that has adaptive value (Catcott, 1975). As such, they must all
be integrated to a greater or lesser extent in any model which seeks to optimally meet the
needs of, and avoid harm to, the animals in our care. These systems include:

Social Behaviour Sexual Behaviour

Feeding Behaviour Parental - Offspring Behaviour
Eliminative Behaviour Comfort Behaviour
Play Behaviour Resting - Locomotory Behaviour
Exploratory Behaviour Agonistic Behaviour

Species-typical behaviours which occur in the domestic environment are co-ordinated to

serve a specific function that has an adaptive value and therefore should not be a
redirected response to some external stressor. For example, spraying is a normal
behaviour in the wild environment, but is a sign of conflict behaviour in domestic
animals kept in close quarters. An awareness of normal behaviour patterns in the species
is essential in order for care-givers to identify and address abnormal behaviour. Both are
extensively addressed by Hart and Pedersen (1991).

In the absence of scientific data to indicate a better management scheme, there is an

underlying presumption that mimicking the wild habitat should be highly desirable for
captive animals in general. Generally, this is accomplished in a modified fashion for most
species, eliminating, for example, such features as predators which the species might
encounter in the wild (Beaver, 1989). However, even in this latter regard, Markowitz and

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Laforse (1987) have discussed artificial prey as behavioural enrichment.

2. Behavioural Enrichment

Behavioural enrichment should, in general terms, foster and promote a full and extensive
repertoire of normal behaviour (ethogram) for the domestic animals, whilst preventing
the development of abnormal behaviour. The provision of physical stimuli and target
objects that will promote the expression of species-typical behaviours should be
incorporated into any plan for behavioural enrichment.

It may be possible to assess program success by the extent to which it prevents

development of behavioural abnormalities and promotes normal behaviour, or minimizes
the expression of or eliminates pre-existing abnormalities that an individual or group
displayed prior to enrichment attempts.

The home range of domestic cats varies tremendously based on population density, need
(hunger), desire (hunting, mating), and such natural and man-made barriers as rivers,
fences, etc. While domestic cats living in rural areas may range over many tens of acres
on a daily basis, as urban crowding intensifies, the territory commonly shrinks to a home
range of one-fifth of an acre each or less (Morris, 1986).

Although cats have been portrayed as asocial loners (Leyhausen, 1990; Beaver, 1981),
some authors now question this contention, as well as how much the social nature is
being changed by selective breeding (Morris, 1986; Liberg and Sandell, 1990; Hurni and
Rossbach, 1989). Even now, most cats are not highly social because they still need
individual space and privacy. However, compatible individuals may share their first order
home (house, room or even chair) as well as their home range (backyard, neighbourhood,
or acres of farmland) (Morris, 1986; Leyhausen, 1990; Macdonald and Moehlman, 1982).

3. Social Peers

Insofar as "solitary confinement" is considered an unnatural condition for most species

(Beaver, 1989), it is necessary to examine the role of conspecifics in promoting well-
being and behavioural enrichment in domestic cats. A wealth of anecdotal information
suggests that pairs and stable groups are successful alternatives to single cage housing for
cats or other species. However, finding scientific data to demonstrate this and other social
functions important to behavioural well-being in cats is a challenge.

Beaver (1981) reports that, although the socialization process is not well understood in
cats, the critical window for their socialization may end as early as nine weeks of age.
Cats weaned at an early age and raised in isolation later displayed excessive undirected
activity, disorganized behaviour and fear of novel situations (Seitz, 1959).

Continued socialization with conspecifics is essential to the well-being of developing

kittens. Blackshaw (1985a) notes that: "Kittens reared in the absence of other cats from

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the 7th week on - and so deprived of the possibility of social play - later showed poor
control of attack and escape behaviours, sexual and parental encounters."

Similar findings regarding social deprivation have been reported for other species
including calves (Broom and Leaver, 1978), rodents (Rosenzweig and Bennett, 1977),
and dogs (Scott and Fuller, 1965).

Cats would be best adapted to the research environment when raised and socialized to
research facilities and human handlers before seven weeks of age. Continued, regular
human contact is also important (Beaver, 1989; Karsh and Turner, 1990) in order to
maintain the human socialization. Beaver (1981) notes that excessive handling can be
stressful to the unsocialized or asocial animal.

Animal caretaker styles can also affect the behaviour of animals (Beaver, 1981, 1989;
Hurni and Rossbach, 1989; Fox, 1986). It is considered that calm, gentle, consistent
keepers reduce the stress in a population.

Visual stimuli can also improve behavioural well-being.

In most animals, changes in routine should be avoided or minimized as much as possible.

For example, even the introduction of a new technician can change liver enzymes in
chimpanzees (Moor-Jankowski and Mahoney, 1989). Hemsworth and Barnett (1987)
report that inconsistent behaviour has the effect of increasing pigs' fear of humans.

4. Enrichment Devices (Artificial Appliances)

Having recognized the need for adequately sized, appropriate caging that is clean, safe,
secure, and suitably bedded, consideration is now being given to provision of species-
appropriate activities through introduction of complexity within the cage space.
Enrichment devices would include, for example, toys, scratch posts, climbing apparatus,
PVC culverts for privacy and play, etc. Activity can be encouraged by hanging an object
that can be swatted or watched, or by providing an object that will roll when batted
(Beaver, 1981).

The concept of novelty is an important one when considering play-articles for cats. It is
widely reported anecdotally that continued exposure to an item reduces the play value to
the extent that the cat quickly becomes indifferent to the article, only to show renewed
interest after a brief period of removal.

One must also consider the age of the cats. Kittens require a variety of articles with which
to play. Play behaviour in kittens occupies almost 10% of total time and is considered to
assist in the acquisition of information and skills includes an opportunity to learn the
communicative value (message and meaning) of displays, particularly "graded displays"
(Blackshaw, 1985a).

5. Food Gathering Activities

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Food gathering activities can be manipulated to foster environmental and behavioural
enrichment. Unfortunately, although much has been written about the food gathering
activities of non-human primates (NHP), there is a relative dearth of literature for
domestic cats. Until recently, it was assumed that if an animal had adequate supplies of
nutritious food and clean water, its "feeding" needs were met. Clearly, this approach
denies the complex range of behaviour exhibited by cats as part of a predator's feeding
behaviour system, which includes, but is not limited to searching, chasing, catching,
killing and consuming prey (food). Four of these five behaviours are redundant for
animals provided with ample nutritious food, and they are deprived of the opportunity to
express such behaviours.

The diet of feral cats includes small rodents, birds, etc., which have partially digested
vegetable material in their intestinal tracts. The cravings many cats have to consume
small amounts of grass, house plants, etc., may reflect a craving for plant material in a
more natural form than as commercial pet food (Leyhausen, 1990; Beaver, 1981;
Blackshaw, 1985b; Beaver, 1980). This need could be met in a variety of ways (e.g., by
providing small amounts of fresh grass or other safe plant, or cooked vegetables which
can be ingested without causing gastric irritation).

A cat's appetite may be affected by lighting and noise level, by the presence or absence of
people, by the type and cleanliness of the container, and by the presence or absence of
other cats (Scott, 1975).

Preference tests have shown that cats prefer their food at 30C (86F) (McKeown and
Luescher, in press). Although it may not be possible or necessary to incorporate this
documented preference in all feeding regimes, such knowledge forms the basis for
enhanced care for individuals undergoing unusual stresses, (e.g., partial anorexia in the
immediate post-operative period), or when introducing new individuals to a group, etc.

Most cats generally do not like to eat from narrow, deep bowls; however, some will only
drink by dipping a paw in such a bowl and licking the foot. If an animal will not eat from
a container on the floor, the bowl may be placed on the perch.

Some cats prefer fresh, clean water that has been allowed to stand for a time to permit the
dissipation of chemical odours that result from water treatment. Other cats refuse to drink
unless from a running source, such as a dripping faucet. Many cats are loath to eat or
drink from any container which is contaminated with odour or saliva from another cat.
Obviously these factors and countless others are significant in providing the optimal
housing for cats under a variety of caging conditions.

Idiosyncrasies of behaviour which reflect the fastidious nature of felines should be

considered in any effort to provide for their social and behavioural needs. For example,
texture is important to cats: the texture of their food can affect appetite; the texture of
resting and sleeping locations can determine preferences in this regard.

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Many cats will not use a litter box that has been soiled by another cat. Indeed, many cats
even have preferences for certain textures or types of litter and will not use others. Even
the location of the food, water, and litter containers relative to each other, resting areas,
doors, etc., can have a significant impact on the well-being of cats.

6. Control of the Environment

Assumptions are widely held that an animal's well-being is enhanced by affording some
degree of control over its environment (Line, 1987). Whether this is expressed in the
negative language of "reducing stress" or the positive language of "environmental
enrichment" we continue to seek ways to allow animals to express their individual needs
or desires. Radio broadcasts played during working hours make cats less frightened by
sudden noises and more easily accustomed to strange human voices (Hurni and
Rossbach, 1989).

The existence of a range of temperatures within the enclosures allows for an individual to
seek preferred resting spots. Provision of safe indoor/outdoor enclosures permits further
freedom of choice and therefore some degree of control. Choice of texture, height,
temperature, and degree of enclosure are examples of a few methods of environmental
enrichment for animals. Cats are noted for their enjoyment of a warm, sunny sleeping
location.

Height (as provided by perches, for example) is also a desirable factor for many cats
when seeking a resting spot (Beaver, 1989; Blackshaw, 1985a). In group housing, access
to the individual's preferred vertical niche, with sufficient elevated resting places for all
cats, will enhance the enrichment program.

Some prefer a dark, secluded spot for rest (Beaver, 1981); others may choose to sleep
closely by another member of the group. Anecdotally, it is noted that some cats prefer to
sleep on (sanitizeable) fleeces and soft blankets.

Many anecdotal observations regarding cats have been well documented in other species.
For example, Chamove and Anderson (1989) report that such arboreal species of
monkeys as the callitrichids rarely come to ground in the wild. Furthermore, they state
that in captivity these monkeys almost never visit a bare floor (1% of the time); however,
the time spent on the ground increases ten-fold if contact bedding such as a leaf-like
substrate is used. Obviously, since the floor may encompass as much as 40% of the total
usable surface area and more than 60% of the horizontal surface area, addressing the
issue of texture preference can provide a potent tool for environmental enrichment for
many species.

Both predictability and controllability are important variables to reduce stress. Therefore,
when controllability cannot be provided, allowing the animal some degree of
predictability is one coping strategy which should enhance well-being (Beaver, 1989).

The concept of predictability must be interpreted in light of species-typical behaviours.

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For example, while some species such as higher primates may respond positively to
changes in time and content of their daily feedings (Line, 1987), other species can
experience unnecessary distress from alterations to their routine. Changes in routine
which may, for example, occur during weekend feeding and cleaning schedules, are
known to be stressful in routine-oriented animals (Beaver, 1981).

7. Housing

The least stressful housing environment for research cats will usually be gang housing,
especially if there are numerous perches on which individuals can rest (Beaver, 1989).
When establishing pairs or small groups, compatibility can first be determined by
observing which animals sit near each other.

Group pens or cages should be provided with adequate, usable, vertical space (e.g.,
shelves, or a tree-like structure with platforms). If kittens are present, access to higher
levels can be provided through the use of a slanting board or similar object. Group
housed cats prefer warmed floor areas on which to sleep (McKeown, Pers. Comm.,
1990).

The housing should provide an area for eating and for elimination. In the latter regard,
provision of a number of litter boxes can reduce the possibility of refusal by individual
animals to use a particular box.

Aggression can occur between adjacent, individually housed animals. Provision of a dark,
secluded hide-out (e.g., box) can allow a retreat from this type of stressful environment
(Beaver, 1981). [This also proved beneficial in NHP which were provided with a privacy
panel (Reinhardt, 1990).]

If animals must be singly housed for an experiment, then where possible and appropriate,
they should be group housed with their original conspecifics between studies. Females
are considered more appropriate for long-term holding, as they are generally more
amiable towards one another and can be kept in groups after several days of limited
exposure (Hurni and Rossbach, 1989).

If possible, all animals not in large pens should be exercised daily, unless contra-indicated
for health or experimental protocol reasons.

Some authors (e.g., Hurni and Rossbach [1989]), suggest that intact males be housed
separately by four to six months unless they remain with litter mates and no stranger is
introduced. However, Taylor (Pers. Comm., 1990) reports long-term success with several
colonies of intact males housed in groups of 6-14. New individuals, carefully introduced,
caused minimal disruption and in more than two years of observation only three to four
episodes of aggression greater than ritual display were noted. None resulted in serious
injury; however, in each of those cases it was only necessary to remove one individual to
restore harmony.

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Asocial individuals should also be singly housed, as the biggest stress to these cats is
other cats. In addition, intact male cats not kept in a breeding harem, cats recuperating
from surgery, and research animals being conditioned to a cage, are usually singly caged;
however, cats can form bonds. This can be illustrated sometimes by the manifestation of
separation anxiety (McKeown, Pers. Comm., 1990).

8. Maternal Behaviour

The duration of gestation in the cat ranges from 60 to 68 days, with an average of 65 to
66 days. During approximately the last third of pregnancy, obvious behavioural changes
occur, although some queens have already been showing increased docility. Along with a
rapid weight gain, due primarily to fetal growth, there is an accompanying increase in
appetite, decrease in activity, and decrease in agility. Distention of the mammae may also
occur.

In the week immediately preceding parturition, the queen will seek a dark, dry area where
she can remain relatively undisturbed. A nesting box fills this need. During this period
before delivery, the queen usually spends an increasing amount of time in self-grooming,
particularly of her mammary and perineal areas. She may also become more irritable or
defensive, possibly as a result of the extreme stress associated with this time of
pregnancy.

As parturition becomes imminent, the female becomes increasingly restless, digs at the
floor or nesting material, and assumes a defecation posture without defecating. There
may be calling vocalizations, especially by Siamese cats, and a few queens become
excessively anxious and even hysterical (Fox, 1974).

Each of the four phases of parturition is highly variable, although their order holds true
for the majority of births. The initiation of each new phase is usually marked by an abrupt
behavioural change, from contractions causing genitoabdominal licking to placental
delivery resulting in the consumption of the placenta (Beaver, 1980).

Hurni and Rossbach (1989) suggest that queens in group housing be provided with a cage
to which they be confined overnight and during mid-day; this would be made available
from just before parturition until four to six weeks after. By letting them out for a few
hours in the forenoon and afternoon into the stock pen, the female remains socialized to
the community and the stress of changes in the population hierarchy is reduced.

9. Random-Source vs. Purposebred Animals

Since it is considered advantageous to house research animals in a social environment,

the housing of animals which are genotypically sociable is a distinct advantage. Cats'
tendency to socialize well is a trait which is carried genetically by the male (McKeown,
Pers. Comm., 1990). Proper selection for genotypically social cats is therefore possible in
purposebred populations. As well, kittens growing up with social conspecifics become
more social than those raised with non-social conspecifics (Schar, 1983). By culling

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animals which exhibit undesirable traits even after adequate socialization (Ringler and
Peter, 1984), the population can be further selected to include more animals which are
behaviourally adapted to the research environment.

For certain types of studies, use of purpose-bred cats provides advantages which enhance
the quality and validity of the research. These include a known health status, and control
over the animal's age, genetic factors, and environment. This allows the production and
use of a much more uniform population of known status. Losses are less, results are more
valid, and therefore fewer animals need be used. In addition, there are many advantages
to the cats kept in the research environment, in terms of their social and behavioural well-
being.

E. DOGS

1. Introduction

Dogs (Canis familiaris) have been human-beings' companions for over 12,000 years
(MacArthur, 1987). In the laboratory, this potential for developing a close relationship
with people may be realized through appropriate socialization of the animal at an early
age. As well, most breeds of dogs used in research, teaching and testing are naturally
gregarious and seek the companionship of other dogs (MacArthur, 1987; Beaver, 1981).
This tendency is also seen in packs of feral or wild dogs travelling together (Dunbar,
1979). Therefore, unless contra-indicated by the protocol, medical condition, or the
animal's aggressiveness, dogs should be paired or group housed with conspecifics in
cages or runs, with space adequate for active normal behaviour. If this is not possible,
dogs should be released at regular intervals into space adequate to permit this normal
species-typical behaviour.

Social rearing of puppies is the most effective means of ensuring compatible conspecific
behaviour as adults (Fox, 1972). Moreover, dogs that have been handled as puppies show
greater resistance to stress and greater disease tolerance than those which are not handled
(Fox, 1975).

The appropriate maintenance of dogs will be discussed under breed differences, criteria
for assessing well-being, housing, socialization to people, and enrichments.

2. Breed Differences

The differences in size between Newfoundlands and Chihuahuas represent some of the
extremes seen among the many breeds of dogs. These differences include not only
morphology, but also temperament (e.g., terriers vs. labrador retrievers), conformation
(e.g., beagles vs. greyhounds), urea metabolism (dalmatians), development of behaviour
patterns (MacArthur, 1987) and other important considerations. Although every dog
belongs to the single species (Canis familiaris), each breed has specific behavioural and
social needs.

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Interbreed morphological differences become important in selection of proper cage size
(even though they have the same body weight, long, lean dogs are likely to require
different cage sizes than short, stocky dogs). The decision to group house will depend to
some degree upon breed differences. Much can be learned appropriate to the well-being
of dogs from a basic knowledge of breed-typical behaviours; however, attention to the
uniqueness of each individual animal is the only way in which well-being can be assured.

In addition to understanding breed differences, an understanding of intrabreed

differences, which are the natural outcome of environmental and genetic factors, is also
of great value. Litter mates, even when reared under the same conditions, may behave
entirely differently.

3. Criteria for Assessing Well-being

Evaluation of animal well-being includes both engineering (environmental) standards

(e.g., minimum cage size, temperature, light cycles, etc.) and performance or outcome
measures (McCarthy, 1989) standards (the dogs' general state of health and their
compatibility in social groups and with people).

The well-being of dogs is dependent on a number of factors which include: training and
dedication of the scientific, animal care, and veterinary staff; a facility in compliance with
this Guide; observations of the animal's physical health (does it appear healthy, alert,
active?); observation of the dog's behaviour; pair or group housing of compatible
animals; and socialization to people.

a) Clinical Observations

i) Eyes

Clarity and expressiveness of the eyes is a good indicator of general health. This should
not be confused with non-eye contact, sometimes demonstrated by dogs raised as
subordinate to people.

ii) Posture

Ill or distressed dogs may appear lethargic or cower in the rear of the cage or kennel.
Abnormal gait or the carrying of a limb is suggestive of a localized trauma or infection.

iii) Hair coat

Ill or chronically distressed dogs will often manifest a rough, unkempt hair coat. Self-
grooming may be absent.

iv) Stool

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Presence of diarrhea, or stool with mucus, blood, or helminths (worm-like endoparasites)
should be questioned.

v) Appetite

Inappetence or too-rapid ingestion of food should be questioned; sudden changes in

weight, drinking or eating behaviour should also be questioned and investigated.

b) Behaviour

i) General

One should look for evidence of how well the dogs are adapted to the environment.
Whether singly or socially housed, they should not normally exhibit highly repetitive or
atypical behaviours. Dogs generally perceive the cage or kennel as home territory and,
when the threat is not too great (e.g., the door remains closed), they may bark in defence
of the territory. Opening the kennel door may elicit very different behaviours; solicitation
from human-bonded animals and fear from unsocialized ones. These differences may not
be seen with the door closed; however, judgement about a fear-response to strange
individuals must be made cautiously, for a certain degree of inquisitiveness or anxiety
over the presence of unknown persons is normal.

ii) Toward cagemates

Compatible cagemates should demonstrate equal desire for attention when the cage is
approached by familiar people. Overly dominant individuals (and dogs overly socialized
to people and undersocialized to dogs) (Beaver, 1981) will prevent subordinate
individuals from being touched by the person, which, at times, may lead to aggression
that continues as long as the person remains at the cage door.

iii) Toward people

Dogs that bark excessively, remain in the rear of the cage, refuse to come to the cage door
even for familiar technicians, or demonstrate aggressive tendencies when approached, are
likely not well-socialized to people. Unsocialized dogs are fearful of people, may become
"fear-biters," are difficult to catch and restrain, and may have physiological variability
incompatible with some scientific studies. These manifestations are de facto evidence of
distress and poor well-being. Such dogs are poor candidates for chronic studies.

iv) Maternal

Toward the end of gestation, the bitch will begin to seek seclusion, a safe, warm, dark and
quiet place, and it is advisable to make such provisions by placing a whelping box in
some corner that she will accept. She should be shown her whelping quarters early on in
her pregnancy and be given amble time to become accustomed to them (Fox, 1972).

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Where possible, during the birth of the pups any kind of handling or interference should
be avoided.

The behaviour of the pregnant bitch changes toward the end of gestation. She is generally
more restless and appears uncomfortable. She may show nesting behaviour and start to
tear up newspapers and to scratch at the floor of the whelping box. Often the bitch will go
off her food, and some occasionally vomit during the few days immediately prior to
parturition. It is not unusual for the bitch to pant a great deal and to regularly look
apprehensively at her hindquarters (Dunbar, 1979).

Nest boxes for whelping bitches, provided with bedding materials and heat sources, keep
homiothermic newborn pups warm and dry.

4. Housing

Housing should facilitate social group formation, human interaction, comfort, and
sanitation. The use of modular cages or runs that can be converted to accommodate either
pairs or groups of dogs is desirable.

Hite, Hanson, Conti et al. (1977) and Hughes, Campbell and Kenney (1989) discuss the
effects of cage size on beagles (the most commonly used purposebred dog). Caging
should permit ready access by personnel and permit visual, olfactory, and auditory
contact with other dogs.

Resting boards made of non-conductive, non-permeable materials should be provided to

permit animals to escape the floor, especially when temperature control and wetting may
be a problem.

i) Social housing

Social housing is desirable for most breeds of dogs. Centuries of interaction with people
and other dogs have developed species-typical behavioural patterns, which must be
understood in order to evaluate and provide for their well-being (Beaver, 1981). Some
breeds of dogs (e.g., hounds) are highly social; others such as terriers are not (Beaver,
1981). Single caging for most social breeds may be stressful.

Movement of an individual(s) away from compatible animals can be disruptive. Dogs

that are removed from a social group (by virtue of health, protocol, or aggression) should
remain in the same room, as close to the same social group as possible, and should be
returned to the group as soon as possible. When the group is stable, positions within the
room should not be changed without cause.

ii) Single housing

For those animals comfortably adapted to solitary living, introducing cagemates may
induce distress. In these circumstances, exceptions to social housing may be appropriate,

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especially where human companionship is provided and they are in visual and auditory
contact with other dogs.

If dogs must be housed singly, they should be in visual, auditory and olfactory contact
with others in the room. It is likely that multiple social groups exist within such a room,
with the most stable groups consisting of individuals immediately adjacent to or across
from each other.

It should be remembered that dominance can be expressed across the aisle, so that an
animal removed from a cagemate because of dominance aggression should not be placed
directly across the aisle from its original cagemate, but moved to a location away from
the overly dominant individual.

5. Socialization to People

Of all the common laboratory species, dogs are the most highly domesticated and adapted
to live in intimate association with people. Socialization creates an attachment and trust
of people, which assists in the development of coping strategies that serve to bridge
periods of adaptation to new procedures and environments, thereby reducing stress and
experimental variability.

Without early exposure to people (i.e., socialization), dogs rapidly become fearful of
humans and manifest fear and distress in a variety of physiological and behavioural ways
(e.g., "fear biting") (Beaver, 1981), all of which are incompatible with their well-being
and can influence the reliability of research data derived from them.

The dog's ability to cope when a person enters the scene, or its environment changes, is a
key criterion to well-being (Dunbar, 1979). Coping connotes the ability of the dog to
adapt to stresses with minimal behavioural or physiological alteration (Archer, 1979).

Therefore, all dogs used in a facility, for whatever purpose, should be socialized to
people, (either in the facility or by the supplier), or serious consideration be given to their
euthanasia or use in acute non-survival studies. Socialization (handling by people) should
take place when pups are between 6-10 weeks of age (Wolfle, 1989a, 1989b; MacArthur,
1987; Fox, 1975). A number of other investigators believe the socialization period should
extend to at least 12 weeks (Pfaffenberger, 1963; Bateson, 1987; Vanderlip, Vanderlip and
Myles, 1985a, 1985b; Scott and Fuller, 1965). Fox (1968, 1990) contends that puppies
deprived of human contact until after 10 weeks of age will be very difficult to handle
later in life.

Adult dogs that demonstrate lack of socialization should not remain in the facility any
longer than it takes to determine that the behaviour is unlikely to respond to remedial
socialization, which, in any event is time and energy consuming and not at all sure of
success (Dunbar, 1979). Such dogs should be either euthanized or used immediately in an
acute, non-survival study. Socialization should be considered a critical part of every
breeding program, and when animals are purchased from a supplier, socialization should

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be written into contract specifications.

Human/dog interactions will ensure continuation of the benefits gained from

socialization. Quantification of contact during the socialization period, in terms of
specific frequencies or durations, is less important than the quality of the interaction.
Puppies are susceptible to attachment to humans or other animals. Thus, repetitive
interaction with people during this period is more important than the exact nature,
frequency, or duration of the interaction.

Wolfle (1990) described the socialization of large numbers of foxhound puppies with
only five minutes per puppy per week. However, it should be noted that this was a
complex, rich, bi-weekly socialization procedure where littermates were treated as a
group and thus each pup benefitted from the interaction of people with the littermates.
Nevertheless, it is clear that the amount of "hands-on" time required to socialize large
numbers of puppies does not seem to be critical, and should be possible to accomplish
with existing staff in most facilities.

Through observation, it should be established whether each dog in a social group is

behaving normally (Beaver, 1981). By having a variety of people participate in the
socialization of each dog and by reinforcing their socialization as adults, the problem of
over-attachment to an individual (person) can be avoided.

Housekeeping routines should include recognition of each dog as the technician works
about the room. Moments taken to speak to and pet the dogs will be repaid through
reduction in the dogs' anxiety and physiological variability (Wolfle, 1990, 1985, 1989a,
1989b). The effects of animal caretaker styles may affect the animal (Fox, 1986) and thus
experimental results.

6. Enrichment Devices (Artificial Appliances)

"Enrichments," often in the form of toys or other appliances, are frequently given to dogs
to produce a desired change in behaviour. For example, abnormal or persistent grooming
may be moderated by giving the dog rawhide or other treats on which to gnaw; however,
this should be done only with the knowledge of the facility manager and investigator.
Beaver (1989) notes that dogs respond well to running through mazes as a means of
environmental enrichment.

Music has long been used to reduce stress in many laboratory animal facilities (Line,
Clarke, Ellman et al. 1987) and dairy barns (Ewbank, 1968), (perhaps because of its
initial stress-reducing effect on the attendant). However, few definitive data exist to
recommend its use for dogs. If used, the volume should be placed at conversational
levels. Levels exceeding 85 db for a sustained period may cause auditory damage. It
should also be remembered that many laboratory animals, including dogs, are able to hear
frequencies above what humans can hear (Dunbar, 1979). If violin music, for example, is
played at high volume, dogs may be in acute discomfort. Conversational (talk show)
radio sound may accustom the animal to the human voice.

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7. Exercise

Exercise for dogs has recently been mandated in American law which requires "that
research facilities shall establish, in consultation with the attending veterinarian, written
procedures and systems for exercise of dogs..." (USDA, 1989).

Dr. Dale Schwindaman, Assistant Deputy Minister for Regulatory Enforcement, Animal
and Plant Health Inspection Services, U.S. Department of Agriculture, has stated that, in
looking at exercise and socialization requirements, it may turn out that social contact with
other dogs or with humans in the case of singly housed animals is more important than
exercise. He reports that it has been proposed that, in addition to housing in compatible
groups, the ability to see and hear other dogs will be required. Singly housed animals
would receive positive physical contact with humans. Any exceptions to the requirement
for exercise and socialization would have to be approved by the Institutional Animal Care
and Use Committee (IACUC). It has also been proposed that animals held in (space that
is) less than what is required for permanent housing as mandated by the Guide for the
Care and Use of Laboratory Animals (USDHHS, 1985) would have to be released for
exercise for at least thirty minutes daily (Schwindaman, 1990).

Scientific data have indicated that cage size had no significant effects on hematologic or
serum biochemical values of purposebred beagles; that the dogs had little inclination to
exercise when released alone into an exercise area, unless humans were present in the
room; and that even a moderate exercise program had no demonstrable effect on
biochemical parameters such as hematology, clinical chemistry or indicators of stress
(Campbell, Hughes, Griffen et al. 1988; Hughes, Campbell, and Kenney, 1989;
Campbell, 1990).

Studies demonstrated that on the average, dogs spend only 0.5 to 1.5 hours daily in any
type of activity, regardless of the housing system. Most of the dog's activity takes place
during the morning hours when there is the greatest amount of human activity in the area.
Providing increased human contact will improve the handling and behavioural
characteristics of the dog, but not its activity, because dogs that do not have enhanced
human contact may move around the cage in an effort to attract attention (Hughes and
Campbell, 1990). These authors contend that they have shown that "dogs are basically
lazy. They do not like to exercise and have no particular inclination to run about an area."
Fox (1986) reports that dogs that are well-fed and content do not exercise routinely.

Although, unlike the U.S., no legal requirements for the exercise of dogs exist in Canada,
the concept of exercise, and perhaps more importantly communal housing and
socialization of the animal, both with conspecifics and humans, is considered of great
importance by the CCAC. Institutions are being asked to furnish documentation of ACC
approval for any dog housed individually. Increasingly, the provision of environmental
enrichment, in its various forms, will be strongly recommended by the CCAC.

In conclusion, it should be remembered that, as Erwin (1985) advised, reactions of

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animals to any type of environmental enrichment should be monitored to determine
whether the desired outcome is achieved.

Beaver (1989) reminds us that studies have not determined the amount of activity that is
actually beneficial to any species. Neither has it been shown whether stereotypic
behaviour is beneficial or harmful (Fox, 1986). Much knowledge of animal behaviour
remains to be garnered and established in order to produce an environment that will
enhance the dog's well-being.

F. NON-HUMAN PRIMATES

1. Introduction

When animals are used, efforts must be made to provide a physical and social
environment conducive to their well-being. As well, social structure makes many
experimental animals sensitive to the ill effects of inappropriate housing conditions. In
Canada, only four species of non-human primates (NHP) are currently used in research,
teaching and testing: rhesus monkeys (Macaca mulatta), cynomolgus macaques (Macaca
fascicularis), African green monkeys (Cercopithecus aethiops) and squirrel monkeys
(Saimiri sciureus). Common and scientific names of a number of species are included as
Addendum 1.

In focusing on NHP, emphasis must be placed on enhancing their social and behavioural
well-being. As Markowitz and Line (1989) point out: "It is clearly possible to find
methods by which environmental enrichment can be combined with a research protocol
to enhance both."

Even though the animal may appear healthy, researchers "cannot be content with
defending the status quo," says Line (1987). He challenges investigators to seek practical
ways to expand opportunities for primates to display normal behaviour, "especially those
housed singly". As Volume 2 (1984) of this Guide noted: "Any primate housed alone will
probably suffer from social deprivation, the stress from which may distort processes, both
physiological and behavioural." It is important, therefore, to provide the company of
compatible conspecifics or other NHP species, and, if this is impossible, increased human
company.

There is a growing body of scientific data on space/cage size appropriate for NHP. While
enclosure size is an important variable, the primary emphasis should be on providing
laboratory animals with the option for species-appropriate activities (Bayne, 1989; Bayne
and McCully, 1989; Line, 1987; Bantin and Saunders, 1989; Fajzi, Reinhardt and Smith,
1989; Chamove, 1989; Markowitz and Spinelli, 1986; Segal, 1989a). Wilson (1982)
found that in captive gorilla and orangutans, enclosure size had no effect on the level of
activity. She suggested that objects within the environment were more important than the
size or complexity of the enclosure. Primates maintained in the absence of external
stimuli tend to display locomotion far more frequently than other categories of behaviour

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such as facial expressions, play, and inquisitive behaviour (Martinic, 1990). Chamove
(1989) notes that many successful enrichment techniques act in a way similar to that of
increasing physical space. Snowdon, Savage and McConnell (1984) note the adverse
effects of too-small caging on reproduction, and the widely accepted fact that small cages
increase the incidence of stereotyped movements and other non-locomotory abnormal
behaviour.

Animals that are not housed properly and treated humanely "yield data that are clearly
confounded with distress" (Markowitz and Spinelli, 1986), i.e., may yield unreliable data
due to the effects of behavioural stress (Levine, 1985) and introduce unwanted variables
(Morton and Griffiths, 1985). It is important, therefore, that those using NHP should first
acquaint themselves with the animal's distinctive characteristics and needs. Differences
within and between species make the task difficult (Snowdon, 1990). Wolfle (1990)
suggests that the researcher consult the psychological literature about animal cognition
and perception. "The best tool of all for providing well-being begins with routine
frequent observation of every animal," he concludes.

2. Interpretation of the Behavioural and Morphological Postures

Primate users sometimes misinterpret the meaning of the behavioural or morphological

signals of NHP, as well as the effect of certain human behaviours on NHP. Inadequate
animal husbandry practices are likely to increase the level of stress during cleaning,
feeding and handling (Fox, 1986; Line, Morgan, Markowitz et al. 1989), and increase the
risk of injuries to both humans and animals. Some of these misinterpreted signals are
described below.

a) The Stare

The stare usually expresses an aggressive mood in NHP (e.g., rhesus). Threats are always
initiated and accompanied by a stare, which usually precedes attack. This behaviour
typically elicits one of the following responses by the recipient: a threat (in increasing
order of intensity are staring back, staring with the mouth open, and grunting), an attack
(lunging, hitting, biting), or a submissive reaction (avoiding to look, leaving, displaying a
fear grimace). Primate users should keep in mind that when looking intensely at a
monkey, they are threatening it and announcing an imminent attack.

b) The Fear Grimace

The fear grimace resembles an exaggerated smile; the mouth corners are fully retracted,
showing all the teeth. This expression may be accompanied by a high-pitched, loud
vocalization (Van Hoof, 1963, 1967). The fear grimace, or bared-teeth display is a
ritualized signal of submissiveness emitted unidirectionally by subordinate to dominant
individuals.

Thus, the fear grimace does not convey a playful mood or an aggressive motivation. The
fear grimace is often inadvertently elicited by primate handlers when they move towards

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a monkey, while looking at it. The best way not to elicit a fear grimace is to avoid staring
at the monkey, and to approach it indirectly.

c) Lip-Smacking or Teeth Chattering

In the many species in which it occurs (e.g., stump-tails), the teeth chattering face
indicates a tendency to flee, the lip-smacking face a stronger sense of social attraction
(Van Hoof, 1963). These are greeting gestures that express an affiliative mood, and
probably include a submissive component, depending on the context.

d) Grooming

Removal of particles of dirt and ectoparasites in NHP is undertaken to establish, maintain

or restore positive social bonds by expressing a state of non-aggression and by reducing
tension. The cleaning function of social grooming is only secondary in importance.
Grooming may pacify another animal, but is also used to maintain social bonds, as in
mothers grooming infants or juveniles, and between members of a mated pair.

Grooming also serves to appease dominant individuals and prevent aggression, to provide
contact-comfort (consolation) to the victims of attacks, to reconcile with an opponent
after a fight, or to reassure subordinates. For example, grooming is directed by males to
females in courtship, and is an important component of co-operative partnerships, such as
coalitions and alliances.

e) Sexual Swelling

In many species, females in estrus manifest a reddening and/or swelling of the perineum.
This signals their sexual receptivity to males. The extent of this swelling is highly
variable among species. Sexual swellings are sometimes misinterpreted as injuries or
symptoms of a pathologic condition. Blaffer-Hrdy and Whitten (1987) present
comparative data on cycle length, duration of menstrual flow, visual signals, and the
behaviour of males as well as females in estrus, for all species.

3. Distinctive Characteristics

a) Locomotion

In contrast with most other experimental animals, which are primarily terrestrial, NHP
are characterized by a number of major morphological and behavioural adaptations to a
three-dimensional arboreal life. These adaptations are stereoscopic vision, manipulative
skills and specific modes of locomotion (climbing, leaping, etc.). Most primates show
vertical flight reactions (Burt and Plant, 1990). For each species there is a defined
behavioural repertoire, and for each species, preferred vertical limits in the wild should
be considered.

b) Social Life

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Most primate species, including the majority of those used in laboratories, are highly
social (Boccia, 1989) and live in complex social groups; however, such social groups are
not necessarily permanent. Species which are primarily solitary include some lemurs and
orangutans (Jolly, 1985). The three major categories of societies are the family, the one
male/multifemale group, and the multimale/multifemale group. Most laboratory primates
belong to the third category.

Many studies have shown that NHP recognize individually every member of their group
and that they establish long-term bonds, extending over years or a lifetime, with many of
their kin and non-kin. Such relationships are bilateral and multidimensional, involving
play, contact-comfort, grooming, sexual activity, protection, support during conflicts, etc.

Because of the social bonding that takes place in most species, social isolation is likely to
affect individual animals. Studies have indicated that the effects of social isolation differ
among rhesus, crab-eating macaques and stump-tailed macaques, with rhesus most
severely affected (Sackett, Ruppenthal, Fahrenbruch et al. 1981).

The major contributions regarding social deprivation were made by Harlow in the 1960s.
Animals raised in total social isolation were characterized as withdrawn, personally
bizarre, and aberrant in social, sexual and exploratory behaviour (Harlow and Harlow,
1965). Goosen (1981) has discussed the isolation of rhesus, noting that individually
housed monkeys have little opportunity to develop coping strategies, and may exhibit
bizarre behaviour patterns (Novak and Suomi, 1988).

c) Cognitive Abilities

Intelligence is reflected in many of the NHP behaviours. For example, lion-tailed

macaques, chimpanzees and capuchin monkeys are known to manufacture probing tools,
and a number of species use "tools" to facilitate food acquisition such as cracking nuts
(Beck, 1980). Research has also indicated that lion-tailed macaques manipulate objects,
and use objects to serve as ladders, to create perches, and to apply leverage (Westergaard,
1988). Macaca nemestrina learn through the observation of others and pass on social
traditions (Cole, 1963). They excel in the use and manipulation of third parties by
forming coalitions and competing for the strongest allies, through the utilization of
affiliative strategies. They are capable of some forms of deception (Smuts, Cheney,
Seyfarth et al. 1987).

d) Emotions

Physical and emotional stress cause the release of various hormones, one of the main
groups of which are the adrenal steroids, particularly cortisol (Moberg, 1985).

If it is accepted that humans and apes are related through evolution, it is considered that
the African apes (the gorilla and two chimpanzee species) are humans' closest kin
(Martin, 1988). NHP exhibit many external manifestations of emotions such as facial

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expressions, vocalizations, postures, gestures, and reactions, similar to those of humans.

Many of the ways in which NHP exhibit emotional responses, for example, separation
and external threat, appear similar to ways in which humans react to comparable
situations. Moreover, many of the abnormal behaviours displayed by captive NHP are
similar to the behaviour patterns of institutionalized humans (Passingham, 1982).

4. Assessing Social and Behavioural Well-being

Psychological well-being can be defined as "a state of harmony, both physical and
psychological of an animal with itself and with its environment" (Coelho and Carey,
1990). Dresser (1988) points out that an animal's well-being "...connotes not only the
absence of pain and distress. It implies as well that an individual's physiological, security
and behavioural needs are fulfilled."

Although we cannot measure psychological well-being in NHP, the following criteria

serve as indications that such a state exists: a) good physical health; b) no signs of pain,
distress or discomfort; and c) no abnormal behaviours.

Moberg (1985) proposes that an animal's well-being is compromised only when it is

stressed by events in its environment, and suggests that researchers should look for
changes in the animal's immune competence, reproductive function or growth and
development: "The existence of pre-pathological states in these systems would indicate
the animal's well-being was threatened."

a) State of Physical Health

Poor health and physical injuries are not compatible with psychological or physical well-
being. Physical health should be routinely assessed by a qualified veterinarian.

Some of the most obvious external signs that can be monitored are the condition of the
coat and skin, the appearance of the eyes, and, if the size of cage permits, the gait pattern.

Examples of abnormalities include unresponsiveness and hypersubmissiveness, hair

pulling-and-eating (Reinhardt, Reinhardt and Houser, 1986), and may include the
crouching posture.

b) Absence of Signs of Pain, Distress and Discomfort

Although NHP express fear through high-pitched screams, when experiencing pain they
are unlikely to emit loud vocalizations. Instead, they display a hunched or crouched
posture, an abnormal or slow gait. They stop self-grooming and avoid conspecifics. They
may moan, refuse to eat and drink, and often attract the attention of conspecifics (Hinde
and Rowell, 1962) (see Control of Animal Pain).

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c) Absence of Abnormal Behaviours

Laboratory primates may exhibit abnormal behavioural disorders in both a barren cage or
in a compatible group (Reinhardt, Reinhardt and Houser, 1986). However, Reinhardt
(1990b) reports that most behave in normal ways even in an impoverished environment.

Every species of NHP is characterized by a specific behavioural repertoire. Ethograms or

descriptive lists of species-typical behaviours, have been published for many species (Van
Hoof, 1967; Bertrand, 1969; Fedigan, 1976; Skinner and Lockard, 1979; O'Keefe and
Lefshitz, 1985; Walsh, Bramblett and Alford, 1982; Erwin and Deni, 1979). A vast body
of information on the social behaviour of NHP living in the wild or in large outdoor
enclosures is also available. For geographical distribution, ecology, diet, reproduction and
social behaviour of a representative sample of taxonomic subgroup of NHP see Smuts,
Cheney, Seyfarth et al. (1987).

Despite the existence of a fair degree of interspecific and intergroup variation, it is

possible to identify general categories of abnormal behaviours observed in captive
species of NHP. Examples are summarized below. More details and a description of
idiosyncratic variants may be found in the literature (Goosen, 1981; Walsh, Bramblett and
Alford, 1982; Erwin and Deni, 1979).

d) Examples of Abnormal Behaviour Patterns

i) Bizarre postures and behaviours

Self-biting, self-clasping, self-grasping, hair pulling-and-eating, feces spreading, face or

eye poking, penis sucking, and "floating limb" accompanied by attack of the limb.

ii) Stereotypical behaviours

Pacing, "saluting," head tossing or weaving, walking or bouncing in place, somersaulting,

rocking, and cage charging.

iii) Appetitive disorders

Coprophagia (ingestion of animal's own feces), urine drinking, hyperphagia (excessive

over-eating), and polydipsia (excessive, long-term thirst).

iv) Abnormal levels of activity

Inactivity, depression.

v) Abnormal social behaviours

Maternal neglect of infants, maternal over-protectiveness of infants, high levels of

fearfulness and over-dependence of infants, inappropriate sexual behaviour,

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hyperaggressiveness, hypersubmissiveness, and avoidance of social interactions.

5. Ways of Promoting Social and Behavioural Well-being

There are a number of ways to alter or improve an animal's environment. Beaver (1989),
for example, suggests five basic means: behavioural enrichment (by creating an
environment similar to the wild habitat), social peers, artificial appliances, food gathering
activities, and control of non-food items. Some of these will be described below.

a) Social Peers

The best psychological enrichment is social enrichment (Crockett, 1990). Providing

opportunities for social interactions is by far the best way to help NHP cope with the two
main categories of problems associated with captivity: boredom (understimulation) and
fear. Social interactions appear to provide the richest source of stimulation and the best
source of emotional security. Segal (1989b), in editing a new publication on NHP noted
that several authors independently reached the conclusion that "the single most important
thing one can do to enrich the life of a captive primate is to provide it with a companion
animal." Moreover, it is believed that more social interaction may take place in an
environmentally enriched milieu (Martinic, 1990).

b) Housing

i) Single housing

The reasons commonly cited for housing primates singly are reviewed, and contested, by
Reinhardt (1990a); these include wounding, disease transmission, dominance hierarchies,
social distress, and undernourishment of a lower-ranking partner. Reinhardt concludes
that chimpanzees, orangutans, rhesus monkeys and stump-tailed macaques have been
resocialized without undue risks or disadvantages: "There is no reason to suspect that
other primate species are less suitable for careful resocialization programs." He warns,
however, that such programs must be tested for each species before social housing is
implemented. Fritz (1989) reported that resocializing singly caged chimpanzees caused
neither wounding nor death.

Single housing is strenuously discouraged, except in those situations when it is necessary

for the experimental protocol, in the case of aggression, or to prevent or contain disease.
When single housing is required for an experimental protocol, the institutional ACC must
be very diligent in assuring this aspect is necessary to achieve the experimental
objectives. ACCs should require the investigator to submit scientific justification for
environmental impoverishment, e.g., as a result of regulations or the demands of the
study protocol.

Singly housed animals have exhibited depressed heart rates and elevated blood pressure
similar to the elevated blood pressure noted in humans diagnosed as being depressed
(Coelho and Carey, 1990).

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If single caging must be used, every effort should be made to enrich it (Reinhardt,
Houser, Eisele et al. 1987; Bayne, Mainzer, Dexter et al. 1991), although strategies for so
doing are apparently still very limited (Chamove, 1989). If possible, NHP should be
given the opportunity to take part in species-typical activities. In the case of singly
housed NHP, the role of the animal care technician takes on added importance (Chamove,
1989; Wolfle, 1990). Familiarity with the handler, surrounding and procedure can
significantly reduce anxiety. Positive reinforcement, using rewards such as food,
encourage animals to accept manipulations without apprehension.

In order to preclude the need for single housing to facilitate sampling of body fluids and
chronic monitoring of physiological parameters, a system of social tethering has been
developed (Coelho and Carey, 1990).

ii) Pair housing

Novak and Suomi (1988) claim that pair-housed primates enjoy a state of physical health
that is usually superior to that of many free-ranging monkeys. Pairing of 700 captive
cynomolgus breeding females (and subsequent offspring) was successfully pioneered in
1983 in the Ottawa colony of the Health Protection Branch of Health and Welfare Canada
(McWilliam, 1989). Although pair housing is not advocated as universally beneficial
(Crockett, 1990; Rupenthal and Walker, 1989), the benefits appear to outweigh the risks
(Crockett, 1990).

Most all age-sex combinations of pair housing are possible. Reinhardt (1987, 1988, 1991)
and Reinhardt, Houser, Eisele et al. (1988) have successfully paired rhesus unrelated
adult females, unrelated adult males, and adults of both sexes with infants.

Pairing provides social stimulation and makes it possible to avoid some of the problems
associated with larger groups (Erwin, 1979; Crockett, 1990). It elicits most species-
typical social interactions for the sexes and ages concerned, except for the multi-animal
interactions.

In an interesting innovation, Reinhardt (1990c) in a recently controlled study provided

isosexually paired (male-male, female-female) adult rhesus monkeys with privacy panels.
It was found that they spent more time in close proximity and more time grooming each
other and huddling, while the incidence of agonistic conflicts was significantly reduced.

Monkeys that are paired should be compatible. Compatibility can be defined as an

affiliative relationship in which such interactions as grooming occur and in which both
members appear relaxed. This only occurs after a dominance relationship has been
established. Reinhardt (1987) suggests compatibility is shown when neither animal
exhibits signs of depression, and neither has inflicted serious injury on the other. Before
being paired, prospective cagemates should be allowed to become familiar with each
other in adjacent cages permitting visual and auditory communication. Dominance
signals, such as staring, open-mouth threat, displacement, or fear grimaces, may be

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displayed in this situation or soon after the monkeys have been put together.

The pair may be formed in a third cage in order to avoid aggression (Erwin, 1979) which
to some species such as gibbons may be related to territoriality. They should be
monitored regularly for signs of incompatibility, such as injuries, avoidance of contact, or
inappetence. Once pairs have been established, they should not be disrupted unless
dictated by an experimental protocol.

iii) Group housing

Larger groups usually offer a much richer social environment and should be favoured
over pairs when groups will remain relatively stable. However, it should be noted that
chimpanzees associate in temporary parties, unlike stable groups of most large primates
(Nishida and Hiraiwa-Hasegawa, 1987). For troop-living primates such as rhesus, the
best way to promote their well-being in the laboratory may be to rear them with partners
or in social groups (Novak and Drewsen, 1989). When groups are being formed,
observers must adjust group composition so the units show minimal aggression (Wolff
and Ruppert, 1991).

There are drawbacks to group housing which should be considered, however. Increased
social interaction may result in disease transmission as well as the risk of injury and death
(Beaver, 1989; Line, Clarke and Markowitz, 1989; Novak and Suomi, 1988; Wolverton,
Ator, Beardsley et al. 1989; Line, 1987). Snowdon (1990) notes that the various species
have different responses to group housing. Group formation may be stressful; Sapolsky
(1989) contends that it takes up to 12 to 15 months for animals' stress markers to return to
normal levels; however, Reinhardt, Cowley, Scheffler et al. (1990) disputes this as
regards to rhesus. Erwin (1979) notes that "fighting is a fairly common occurrence in
primate groups even in natural settings, but trauma due to aggression is an especially
pressing problem in captive groups of macaques and baboons."

NHP are prompt to form coalitions through which they establish their dominance ranks
and compete for food and sexual partners. Removing a monkey from its group may
disrupt the existing network of alliances and induce rank changes, which may be
associated with vicious fighting, resulting in injuries (Kaplan, Manning and Zucker,
1980; Reinhardt, Reinhardt, Eisele et al. 1987). Animals that are to be reintroduced
should be kept away from the group for as short a time period as possible.

c) Social Interaction with Humans

It is suggested that there be as much interaction as possible between the NHP and the
investigator or technician (Hearn and Dixson, 1984; Bayne, 1989). The interaction,
however, must not involve handling other than what is necessary for the maintenance of
the animal or for investigational procedures. The necessary precautions are described in
Volume 2 of this Guide (CCAC, 1984).

Direct physical contact between humans and NHP should be evaluated from facility to

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facility. In many instances it should be kept to an absolute minimum for example,
because of the need to break the human/animal bond when staff changes occur or when
an animal must be euthanized, as well as the hazards posed by zoonotic diseases. Some of
the most significant diseases are Herpesvirus simiae infection (B-Virus) and infectious
hemorrhagic fever viruses. Also, many NHP have extreme physical strength in
relationship to body size, and can inflict serious injury on personnel. Furthermore,
humans can transmit infectious diseases to primates, e.g., measles, tuberculosis.

Forced physical contact between humans and NHP can be extremely stressful to the
monkey. Moor-Jankowski and Mahoney (1989) have reported that even the introduction
of a new technician can change the NHP's liver enzymes to the point that it can
compromise a study. Many animals react to the presence of a human observer with anti-
predator behaviour such as mobbing and alarm calling (Caine, 1989), and threat
behaviour toward observers (Wolff and Ruppert, 1991). Talking to the monkeys,
combined with the physical presence of the human, will accustom the NHP to the human
presence, and may thus reduce stress. Burt and Plant (1990) suggest that use of mesh
cage fronts are preferable to the barred variety aiding interaction between animals and
staff.

d) Diet Supplementation and Food Gathering Activities

The core diet of the NHP should be a complete, well-balanced commercial diet, or
alternatively a diet of equal quality prepared in the diet kitchen of the institution. This
diet should be supplemented to suit the nutritional requirements of the primate species
being used (Jones, 1972).

Supplementation and innovative ways of presenting the food to the monkey are effective
ways of enhancing well-being, particularly of those in individual caging. Examples of
items used for supplementation are raisins, fruit, chicken scratch feed, and Prima-Treats
(Addendum 2). Fresh branch clippings may be used for supplementation, providing toxic
plants are avoided and the dust and pesticides are washed off the branches.

The diet supplement can also be provided as food puzzles (e.g., Kong Toys) containing
frozen juice, peanut butter or raisins (Addendum 2). Seeds, etc., can be hidden in deep
litter. These methods of supplementation require the monkey to search and/or work for
the food (Anderson and Chamove, 1984). This challenge will simulate the foraging
activity which the NHP pursue in their natural habitat, and has been found to reduce
stereotypies and increase exploratory behaviour (Anderson and Chamove, 1984; Boccia,
1989).

e) Exercise

The majority of NHP in their natural environment move widely and regularly within their
established home ranges. Except for owl monkeys and many prosimians, primates are
diurnal, spending large portions of the day foraging for food or participating in grooming
and other social activities. For this reason, NHP housed in standard cages over long

138
periods of time, whether they are held singly or in pairs, appear to benefit from species-
appropriate activities reminiscent of those in the wild (Hearn and Dixson, 1984;
Chamove, 1989; Burt and Plant, 1990).

It has not been demonstrated that simply enlarging the amount of available space will
improve the well-being of the animal (Novak and Suomi, 1988; Fajzi, Reinhardt and
Smith, 1989; Novak and Meyer, 1988). Indeed, an increase in aggression has been
associated with an increase in space for some captive primates (Novak and Meyer, 1988).

Exercise cages for NHP were introduced over a decade ago (Tolan, Malone and Rogers,
1980). However, it is considered that environmental complexity, rather than size alone
should be increased (Line, 1987; Line, Clark and Markowitz, 1989; Bryant, Rupniak and
Iversen, 1988). Wolff and Ruppert (1991), reporting on an exercise program involving
rhesus, cynomolgus and capuchins, note that the majority of animals interacted in a
positive fashion. Constant observation prevented fights and thus minimized injury. Much
of the aggressive behaviour was non-physical (e.g., vocalizing or teeth baring rather than
biting).

The NHP entering an exercise area for the first few occasions will usually exhibit a fear
response (Wolff and Ruppert, 1991). However, exercise can be stimulated by
synchronizing it with feeding. Deep litter in which food can be hidden can also be used in
the exercise cage and will stimulate the monkeys to forage. To instill a feeling of security,
freedom of movement between the exercise cage and the home cage is preferable.

If more than one monkey is exercised at the same time, the animals should be cage mates.
However, it is sometimes possible that singly housed or paired monkeys can be exercised
in larger groups if they have been confined in the same room, and are exercised together
on a regular basis. These animals must, however, first be tested for compatibility.

f) Physical Enrichment of the Cage Environment

It is important that the animal be given as much control (or even the perception of
control), as possible over its environment (Line, 1987). Guidelines for minimum cage
size have been established by the CCAC (see Appendix I). Life in cages can be enriched
and activity promoted by the installation of devices such as small branches (O'Neill,
1989), toys (Line, Clarke and Markowitz, 1989), perches (Crockett, 1990), swings
(Bayne, Suomi and Brown, 1989) and food puzzles (Beaver, 1989; Chamove and
Anderson, 1989). Such enrichment is particularly important to the singly caged animal
(Fajzi, Reinhardt and Smith, 1989), where devices which encourage foraging appear most
successful (Crockett, 1990; Bayne, Mainzer, Dexter et al. 1991). Jerome and Szostak
(1987) contend that foraging devices are used more frequently than play objects by
baboons. Climbing is an especially good exercise. Bryant, Rupniak and Iversen (1988)
contend that enrichment of the home cage could benefit the animals more than exposure
to a playpen routine. Enrichment devices and their suppliers are listed in Addendum 2.

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Wolfle (1990) notes that choice tests permit the animal to indicate that one environment
or toy is preferred over another. Other tests measure the frequency of use of new space or
new "toys." It has been suggested that toys be rotated in order to minimize
understimulation (McWilliam, 1989).

Because of the importance of vision to the NHP, particularly M. nemestrina, (Cole, 1963),
cages should be positioned so that the monkeys can see animals of like species. Solid-
sided caging prevents visual contact. If physical contact is possible, there must be
assurance that the animals are compatible.

There is a diversity of opinion with respect to the use of audiovisual devices (radio,
video, television) as a means of enhancing the well-being of NHP. They appear to be of
most benefit if the monkey can turn the equipment on and off at will (Beaver, 1989; Line,
Clarke, Ellman et al. 1987). In some situations, audio may serve as a contentment device
for the primate; however, it is possible for some sounds to be irritating and stressful.

Visual means of enrichment may be stressful to the animal if the monkey perceives the
picture to be threatening. This may be circumvented by the preparation of tapes or videos
especially prepared for primate entertainment. It has been reported anecdotally that
monkeys are particulary fascinated by visuals depicting their natural environment, or
animals that are found in their natural habitat. NHP are fascinated by videos of
themselves (Chapais, Pers. Comm., 1990).

6. Disposition

Following completion of the study, consideration should be given to further use of NHP
utilized in non-invasive research, in an effort to minimize the numbers of animals used.
However, monkeys used in invasive or stressful projects should not be subjected to
further stressful procedures or conditions and should be humanely killed according to
CCAC euthanasia guidelines found elsewhere in this Guide. Maximum utilization of
NHP tissues, histological specimens, etc., is encouraged.

Following completion of research, retention of an animal in the laboratory, on the

assumption that it may be required for future studies, is rarely justifiable.

7. Summary

In considering all the factors related to well-being of NHP, we should remember that
"because well-being is subject to past experiences, present circumstances, and future
expectations, it is a dynamic and changing phenomenon" (Wolfle, 1990).

G. RODENTS AND RABBITS

1. Introduction

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Over the course of the past 25 years, standards have been established which are
considered to represent optimum housing requirements for laboratory animals. Many of
the improvements in housing and management standards were primarily designed to
reduce variables and enhance the reproducibility of experimental results (Lang and
Vessell, 1976). However, during the past decade, increased emphasis has been placed on
the behavioural and social needs of animals kept in the laboratory setting. Most
investigations into enhancement of the animal's environment have concentrated on the
"higher mammals," particularly NHP. This document's conclusions have been based on
contemporary concerns.

Laboratory rodents and rabbits are frequently perceived to have relatively few
requirements other than basic housing, husbandry, and dietary needs. Thus, control of the
environment has frequently been the primary consideration, with little (or no) emphasis
on other areas.

"Well-being" is frequently difficult to assess objectively in these species. Weight gains,

general behaviour, and adrenal weights are examples of criteria which have been used in
studies of this type (Chamove, 1989).

Housing conditions should be evaluated carefully for each species, and consideration
given, wherever possible, to innovative group housing in species such as guinea pigs and
rabbits.

Qualified investigators should be encouraged to do additional objective, controlled

studies on the environmental needs and preferences of rodents and rabbits in the
laboratory setting.

Awareness of normal behavioural patterns in each species is essential. For example,

coprophagy (reingestion of feces) is a normal function in several species, including
rabbits and rats (Smelser, 1985; Newton, 1978). Rats normally ingest 35-65% of their
feces on a daily basis. Deprived of this opportunity, 15-25% reductions in weight gains
have been observed (Newton, 1978).

Beaver (1989) suggests that five aspects in particular may contribute to environmental
enrichment: behavioural enrichment, social peers, artificial appliances, food gathering
activities, and control of the environment. Some of these are discussed below.

2. Behavioural Enrichment and Social Peers

Social interaction with peers is recognized to be a desirable, if not an essential aspect of

laboratory animal well-being.

a) Mice

Mice thrive when housed in groups of two or more per cage. In one study, evidence of
"stress" was minimal in mice housed at four per cage, compared with groups of two or

141
eight per cage (Peng, Lang and Drozdowicz, 1989). A high incidence of stress-related tail
lesions has been observed in cages housing up to 40 mice which were placed together
after weaning. The problem was resolved when the groups were reduced to five per cage
(Les, 1972).

As another example, female C3H/He mice in an intensive breeding program, and housed
under conditions of severe social stress, had an incidence of spontaneous mammary
tumours considerably different from counterparts kept under ideal conditions. At 400
days of age, approximately 90% of animals maintained under adverse conditions had
mammary tumours, while the incidence of tumours was around 10% in females housed
and mated under optimum conditions (Riley, 1975).

Compatibility is a critical consideration. It may be impossible to house male mice

together after puberty, particularly those of more aggressive strains.

b) Rats

Rats are frequently housed singly for certain types of studies; however, it is desirable that
two or more compatible rats be housed together in an appropriate cage. Post-puberal
males are usually compatible, particularly if they have been together since an early age. It
has been shown that even groups of highly standardized male rats exhibit a high level of
variability of behavioural patterns (Grtner, Ziesniss, Karstens et al. 1991).

c) Guinea Pigs

Guinea pigs live in groups of five to ten individuals in the wild (Sutherland and Festing,
1987) and thrive under group housing, although it is unlikely that two or more sexually
mature males will live together without incident unless they have been together since
birth. In their natural environment, guinea pigs exhibit a strong herd or family orientation,
and this should be maintained in the laboratory setting, if at all possible. The one boar per
harem arrangement is the recommended procedure in breeding colonies. Guinea pigs
should not be housed singly; however, if this is necessary, Sutherland and Festing (1987)
recommend a minimum of 700 cm2. Vocalization appears to play an important part in
guinea pig social behaviour, and they call for attention from human caretakers
(Sutherland and Festing, 1987).

d) Hamsters

Adult hamsters are frequently caged separately because of their tendency to fight, with
the exception of the female during the time she is ready to mate. However, they have
been housed together under certain circumstances, particularly if they have been weaned
and raised together since birth (Hobbs, 1987); nonetheless, the European hamster
becomes more aggressive as it grows older. Hamsters spend more time in social
proximity if they have had prior group housing experience. Singly housed hamsters show
more agonistic behaviour with conspecifics, and lower weight gains than group-housed
animals. Early housing experience can profoundly affect later social preferences and

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behaviour.

e) Gerbils

Most gerbil species are gregarious and live in large groups (Norris, 1987). Therefore,
they should be housed in pairs or in larger groups wherever possible. If animals are
placed together before they reach puberty, fighting should not be a problem. Gerbils
usually mate for life; thus, it is advisable that single pairs be kept together throughout
life. Most are normally docile, although aggression may be noted after pairing for mating.

Mature mongolian gerbils of either sex may show a characteristically severe form of
epileptiform seizure (Norris, 1987).

f) Rabbits

In their natural habitat in the wild, rabbits of the genus Oryctolagus are social animals,
frequently living in warrens of up to 100 or more rabbits of various ages. In the
laboratory, convention has dictated that sexually mature animals be housed singly: a) to
avoid fighting injuries; and b) to prevent ovulation and subsequent pseudopregnancy due
to physical interaction in mature does. Male rabbits, if penned together, become
increasingly aggressive from about 90 days (Adams, 1987). However, group housing for
adult rabbits has been under study, both in the laboratory setting, and in commercial
rabbitries (Stauffacher, 1992; Love, 1988; Anon., 1989a).

Group housing in larger enclosures has provided animals with the opportunity to live a
more natural lifestyle, including ample opportunity for adequate exercise, mutual
grooming, and general improved well-being (Love, 1988; Boyd, 1988). Breeding
colonies have been established, using the group housing approach (Anon., 1989b). In
some facilities, compatible rabbits are allowed to exercise in a designated floor area
several times per week.

3. Enrichment Devices (Artificial Appliances)

a) Mice

Mice have used empty plastic water bottles placed in the cage for a "urinal," and an
additional bottle for nesting and as a "bolt hole". It was concluded that provision of the
bottle was beneficial in several respects, including improved sanitation, and an
opportunity to establish their own optimal environment in the nesting bottles (Boyd,
1988). However, in one study, the addition of objects such as flower pots and bricks has
been reported to increase aggression among male mice (Ayling, 1989), presumably
because of territorial instincts.

b) Rabbits

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The use of resting boards has been shown to have a calming effect on rabbits, which use
them to hide beneath (Anon., 1989a), and the use of tubing as "bolt holes" has been
suggested.

c) Hamsters

Hobbs (1987) states that wheel-running has not been shown to be beneficial; however, a
raised top to the cage provides opportunity for climbing and exercise.

d) Gerbils

It has been recommended that gerbils be supplied with appliances such as PVC plastic
pipe since they are burrowers in their natural habitat, and retain this behavioural pattern,
as well as food hoarding, in the laboratory (Norris, 1987). It is likely that other small
rodents would also benefit from these additions.

4. Caging and Bedding

i) Floor space per animal is an important consideration, and requirements for individual
species have been identified in Appendix I [the U.S. and the U.K. have developed
standards as well (USDHHS, 1985; UFAW, 1987)]. Although ample floor space per
animal is essential, there is some evidence that the actual needs for group-housed guinea
pigs, for example, may be less than current guidelines indicate (White, Balk and Lang,
1989).

ii) Solid bottom cages are strongly recommended for housing rodents, particularly for
long-term studies. Solid floors with appropriate bedding are particularly critical for
breeding rodents (Weihe,1987). Wire bottom cages, although less labour intensive to use,
are far removed from the natural environment.

iii) Bedding is also an important consideration. For example, gerbils are active
burrowers, and prefer bedding that can be used for digging and tunnel making, an
important activity in this species. There have been studies of bedding preferences in small
rodents (Iturrian and Fink, 1968). Straw bedding has been recommended for rabbits
(Adams, 1987) who also notes that breeding females kept in metal cages must be
provided with nest-boxes some days before parturition. In metal caging, Adams (1987)
found that 16 mm mesh, 2 mm gauge wire was satisfactory in preventing sore hocks.

a) Rats

It is now recognized that rats like to run, stand on their hind legs, and jump (Weihe,
1987); unfortunately, presently available caging does not permit this. Weihe (1987)
recommends addition to the caging of paper, wood, pellets or grain as a means of
environmental enrichment. He also suggests caging of solid plastic with a wire mesh lid,
and criticizes use of wire mesh-floored cages. Rectangular cages are more satisfactory
than square caging, with 20 cm high cages suggested (Weihe, 1987).

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b) Mice

In one study conducted in mice, vertical dividers were placed in cages, and the animals'
performance and well-being compared with that of animals housed in conventional cages.
Mice preferred the complex cages, and appeared to be "less emotional" than were the
mice kept in regular cages. It was concluded that the divided cage represented a more
natural housing arrangement, and that its use would lead to healthier animals (Chamove,
1989).

c) Gerbils

Any cage suitable for rats and golden hamsters is satisfactory for gerbils. As gerbils often
stand erect on their hind legs, the cages should have a solid bottom, with floor to lid
height at least 15 cm. A monogamous breeding pair requires a floor area of about 700-900
cm2 and gerbils caged in large groups need about 100 cm2 floor area per animal (Norris,
1987).

d) Rabbits

Adams (1987) suggests that, for laboratory purposes, rooms designed to accommodate
units of 50-60 rabbits are best. If metal caging is used, 16 mm mesh, 2 mm gauge wire is
satisfactory in order to preclude sore hocks.

These are often built with portable sides with or without a roof or raised grid floor. They
may be used to house a variety of species such as cats, dogs and NHP. Rabbits and guinea
pigs have also been successfully housed in floor pens. In Swiss studies, near-to-nature
surroundings for rabbits have been replaced by manageable artificial substitutes
(Stauffacher, 1992).

e) Guinea Pigs

In guinea pigs, easily sanitized boxes with an end opening, placed in the floor pens, have
proven to be an unqualified success. These boxes serve as a place to hide and as a secure
place for farrowing, and provide some variety in the environment (White, Balk and Lang,
1989).

5. Food Gathering

Good quality legumes or appropriate vegetables (e.g., carrots, cabbage, etc.) are useful
supplements to commercially available diets for guinea pigs, rabbits and gerbils. Seed
mixtures are recommended additions for species such as gerbils and hamsters although
Norris (1987) warns that gerbils will eat sunflower seeds and exclude other seeds. He
also suggests feeding seed mix to young animals on the cage floor. Nutritious food items
of this type will provide a pleasant diversion, as well supply additional nutrients to these
species. However, quality control of such materials is essential since there is the potential

145
for biological or chemical contamination. This practice could be contra-indicated in
animals in nutritional or toxicology studies.

Rabbits are said to prefer pelleted commercial feed rather than meal, and to have a higher
requirement for fibre than other species (Adams, 1987).

6. Control of the Environment

Ambient temperature, humidity, air changes, frequency of cage cleaning, light-dark

cycles, noise and daily routines, are examples of environmental conditions that affect the
welfare of animals in a research facility (Clough, 1982; Gamble, 1982; Riley, 1975;
Peterson, 1980; McSheehy, 1983; Everitt, McLaughlin and Helper, 1987; Besch, 1980;
Grtner, Bttner, Dhler et al. 1980; Anon., 1989b).

Significant variations in certain blood constituents were observed in rats subjected to

various handling and experimental procedures. On the other hand, the presence of a
familiar animal attendant in the room in the absence of manipulations had minimal
influence on the blood characteristics under study (Grtner, Bttner, Dhler et al. 1980).
Sudden changes in humidity may adversely affect rabbits (Anon., 1989b).

In laboratory rodents and rabbits, stimuli and conditions have been identified which may
have adverse effects on psychological well-being and general health. Animal rooms may
not be used for performing any experimental procedures requiring manipulation,
particularly those likely to evoke fear and/or vocalization.

a) Noise

Levels of 50-70 DBA or higher are considered likely to be detrimental to the hearing of
rodents and rabbits. Adverse effects have included audiogenic seizures in young mice
(Bevan, 1955; Gamble, 1982), and reduced fertility in mice and rats (Newton, 1978).

b) Lighting

Light intensity can influence rodent activity, maternal behaviour, and various other
aspects of reproductive physiology (Clough, 1982). Reproductive disorders have been
identified in mice and rats housed under inappropriate light/dark cycles, or in the absence
of such a cycle (Newton, 1978). In albino rats, continuous exposure to light levels of
greater than 700 Lux can cause severe retinal degeneration over a period of time (Everitt,
McLaughlin and Helper, 1978; Clough, 1982; Semple-Rowland and Dawson, 1987), and
there are other reports of light-associated retinal damage in albino rats (McSheehy, 1983).
Data on acceptable light levels for laboratory rodents are available (ILAR, 1977).

H. WILDLIFE HELD IN THE LABORATORY

Wildlife species which are threatened, endangered or Convention on International Trade

146
in Endangered Species of flora and fauna (CITES)-listed must be conserved, and every
effort should be made to replace these animals after study, either through reintroduction
to the environment of origin, or placement in captive breeding-release projects.

Researchers planning to use large numbers of animals should, where feasible, breed
replacement stock rather than continuing to remove animals from the wild.

The chapter on wild vertebrates in Volume 2 of this Guide (CCAC, 1984) is

comprehensive and wide-ranging, and should constitute the primary source of
information and guidance, along with the Categories of Invasiveness and Ethics of
Animal Investigation document found elsewhere in this Volume.

As noted in Volume 2, such animals should only be brought into an institution after the
investigator proposing to use them has demonstrated adequate knowledge of the animals'
social and behavioural requirements or those of a closely related species. Those who will
be responsible for such animals must also be able to provide for appropriate management
and housing before the animals are introduced into the laboratory.

A number of recent, excellent publications are listed under Additional Reading.

I. REFERENCES

ADAMS, C.E. The laboratory rabbit. In: Poole, T., ed. UFAW (Universities Federation
for Animal Welfare) handbook on the care and management of laboratory animals. 6th
Ed. Harlow, Essex: Longman Scientific and Technical, 1987: 415-435.

AGRICULTURE CANADA. Publication 1771/E. Recommended code of practice for the

care and handling of pigs. Communications Branch, Agriculture Canada, Ottawa, Ont.,
K1A 0C7. 1984.

IBID. Publication 1898/E. Recommended code of practice for the care and handling of
farm animals--pigs. In Press.

IBID. Publication 1821/E. Recommended code of practice for the care and handling of
special fed veal calves. 1988.

IBID. Publication 1757/E. Recommended code of practice for the care and handling of
poultry from hatchery to processing plant. 1989.

IBID. Publication 1853/E. Recommended code of practice for the care and handling of
dairy cattle. 1990.

IBID. Publication 1870/E. Recommended code of practice for the care and handling of
farm animals--beef cattle. 1992.

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 ADDENDUM 1

COMMON AND SCIENTIFIC NAMES OF NON-HUMAN PRIMATES

Common Name Scientific Name

African Green monkey Cercopithecus aethiops

Assamese macaque Macaca assamensis
Baboon Papio spp.
Bush baby Galago spp.
Capuchin monkey Cebus apella
Chimpanzee Pan troglodytes
Common marmoset Callithrix jacchus
Cynomolgus macaque Macaca fascicularis (M. irus)
Gibbon Hylobates spp.
Japanese macaque Macaca fuscata
Lion-tail macaque Macaca silenus
Owl monkey Aotus trivirgatus
Rhesus monkey Macaca mulatta
Squirrel monkey Saimiri sciureus
Spider monkey Ateles spp.
Stump-tailed macaque Macaca arctoides (M. speciosa)
Tamarin Saguinus spp.
Pig-tail macaque Macaca nemestrina

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 ADDENDUM 2

SOURCES OF ENRICHMENT DEVICES

Puzzle feeders and treats Primate Products
Kong Toys 1755 East Bayshore Road, Suite 28A
Prima-hedrons Redwood City, CA 94063
(perch and swing toys) Telephone: 415-368-0663 Fax: 415-368-0665
Nyla Balls H & L Pet Supplies
(Wolf size are used for 27 Kingston Crescent
Cynomolgus monkeys) Kitchener, Ontario N2B 2T6
Telephone: 519-743-8954

Central Sales
60 Eastern Avenue
Brampton, Ontario L6W 1X8
Telephone: 1-800-387-2522

Rolf C. Hagen Inc.

3225 Sartelon
Ville St. Laurent, Quebec H4R 1E8
Telephone: 514-332-0914
Fruit-punch flavoured P.J. Noyes Company Inc.
fructose pellets (for Whitefield Road
environmental enrichment treats) P.O. Box 381
Lancaster, NH 03584
Telephone: 603-788-4952
Fleece foraging/grooming board BIO-SERV
Foraging crumbles "mixed size" P.O. Box 450
Turf foraging board Frenchtown, NJ 08825
Foraging bites Telephone: 908-996-2155
Prima-burgers Outside USA: 908-996-4123
Prima-gel
Prima-treats
Marmoset diets
Prima-treats Kaplan Laboratory Animal Supplies and
Prima-Foraging/Grooming Services
boards 4960 Almaden Exp., Suite 233
Kong Toys San Jose, CA 95118
Telephone: 408-266-1235
Nyla Balls and Nyla Toys
Boomer balls

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 Chapter VII - Special Practices

VII. SPECIAL PRACTICES

A. ANIMAL ACQUISITION

1. Procurement

All animals must be legally acquired. Animals received as donations for research should
have been legally released (see Laboratory Animal Care, Identification and Records). It is
almost always preferable to obtain standard laboratory species from an established
breeder or licensed supplier. Several provinces have regulations governing the
procurement of dogs (see also Responsibility for the Care and Use of Experimental
Animals). In Ontario, laboratory animal supply facilities are licensed and inspected. All
commercial producers of laboratory animals, regardless of whether or not they come
under provincial legislation, are expected to provide housing facilities and to follow
practices similar to those outlined in this Guide.

It is in the best interests of the user and the supplier to co-operate in the elimination of
any undesirable condition affecting the health and quality of the animal. The institution
(i.e., the receiver) should inform the supplier of any undesirable conditions observed in
the stock received. The supplier should, if requested, provide detailed information on
health status monitoring, breeding, and husbandry practices followed.

The acquisition of animals should be dependent upon the prior approval of the project by
the institutional Animal Care Committee (ACC). Acquisition procedures should be in
place to ensure that the institution will have an up-to-date and ongoing inventory of all
animal experiments for which it may be accountable, and to allow for the prior
preparation of appropriate space and such other arrangements as may be necessary for the
reception of the incoming animals.

2. Transportation

a) Introduction

Depending upon the species and size of the animal, the modes of transportation can be by
land, sea or air. For most laboratory species, the most common method is either by
ground transportation, over relatively short distances, or by air for longer distances. The
objective of any method of travel is to ensure the safety, security, and comfort of the
animal in the container, and to take it to its destination as quickly and as safely as
possible.

Although many agencies are concerned with animal transportation per se, those dealing

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with the transportation of experimental animals are few in number. The Animal
Transportation Association (formerly known as the Animal Air Transportation
Association) (AATA) is involved in making improvements to all modes of transportation.
Publications are available on the subject of animal transportation. The Canadian
Federation of Humane Societies (CFHS) (1988), for example, conducted a survey of
surface livestock transportation in Canada. A recent British Veterinary Association (BVA)
symposium has examined the welfare of animals in transit (Gibson, Paterson and
Conville, 1986), as has a symposium of the World Association for Transport Animal
Welfare and Studies (WATAWS) (Laing, 1991).

b) Regulations--Containers and Transportation

The International Air Transport Association (IATA) annually produces the IATA Live
Animal Regulations, which includes information concerning the documentation, the
containers and other requirements for humane transportation of live animals (IATA,
1992). Some 81 containers are described under the headings of species, design and
construction, preparations for dispatch, feeding guide, general care and loading. While
the container information is specific for air transportation, the requirements for the
containers are applicable to all modes of transport, for they ensure safety, comfort, and
security for the animals (Rowsell, 1992).

In order to ensure accurate technical content, the IATA Regulations are prepared in
consultation with representatives from the Convention on International Trade in
Endangered Species of Wild Fauna and Flora (CITES) and the Office International des
Epizooties (OIE). For over 10 years, the Canadian Council on Animal Care (CCAC), the
International Council for Laboratory Animal Science (ICLAS), as well as the Eurogroup
for Animal Welfare, have maintained close liaison with the IATA Live Animals Board,
addressing primarily the issues concerned with transportation of experimental animals.

Legislation in Canada addressing transportation includes the (federal) Health of Animals

Act (C-66, June, 1990, rev. March, 1992; 38-39 Elizabeth II, Chapter 21), Ontario's
Animals for Research Act (Revised Statutes of Ontario, 1980, Chapter 22 as amended by
1989, Chapter 72,s6 and Regulations 16,17,18,19), and, in Alberta, the Universities Act
(Section 50, "Dog Control and Procurement"; Regs. 341-366). In 1972, Alberta
Regulation 33-72 was expanded to cover the treatment of animals (Rowsell, 1974). As
well, there is specific legislation governing the transportation of animals in provinces and
specific bylaws in municipalities.

The transportation of animals from the United States into Canada is affected by the U.S.
Department of Agriculture's Animal Welfare Act (1966) by specifying minimum and
maximum temperatures to which animals in transit may be exposed. The U.S.
Department of the Interior, U.S. Fish and Wildlife Service, is responsible for the
enforcement of the importation of wild mammals and birds; as well, the U.S. Department
of the Interior is responsible for the enforcement of the Marine Mammal Protection Act.

c) Transportation Stress

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One of the least controllable variables in animal experimentation is the effect on the
research caused by moving animals from one area to another (Landi, Kreider, Lang et al.
1982, 1985; Aguila, Pakes, Lai et al. 1988; Bean-Knudsen and Wagner, 1987, Reinhardt,
1992). This might involve great distances between countries, or minor distances, such as
within the animal facility itself. Even moving animals within the animal room affects the
stress indices, which are more greatly influenced if the individual doing the transporting
is a stranger (Grtner, Bttner, Dhler et al. 1980).

The stressors in animal transportation include inexperienced handlers, the amount of time
spent in preparation, in transit, and on arrival at the destination, and the state of the mode
of transport, e.g., rough roads, rough rail beds, rough seas and air turbulence. Of
importance are the comfort and suitability of the container, sufficient time spent for
adaptation to the container prior to transport, and the temperature and ventilation of both
the container and the ambient temperature of the environment and the various
temperature zones through which the animal may pass. It is essential to make appropriate
pre-arrangements concerning the transport of the animal in order to minimize the length
of time spent in transit.

Transportation stresses can be minimized by avoiding long distance shipping, slow

modes of transport and, as mentioned, by acclimatizing the animals to the containers and,
if possible, to the mode of transport. Kiley-Worthington (1990), in studies involving the
transportation of animals in circuses and zoos, showed that unhandled, naive animals
(i.e., those that are wild or have not been domesticated or are not used to confinement),
will probably suffer the most. Although there is a great deal of concern about circus
animals, show dogs, or competitive horses subject to regular transportation, this
researcher reported that there was "no evidence suggesting that the transporting of circus
animals is necessarily or even unusually distressing or traumatic for the animal, although
it is for naive livestock."

The requirement for training those involved in the transportation of animals is often
neglected. Although it is well established that inexperienced handlers can affect the
animals significantly, little has been done to ensure proper training of individuals
engaged in animal transportation. In 1978, the CCAC produced an audio-slide
presentation on "Humane Transportation of Live Animals" which was made available to
all airlines and any of the other agencies engaged in the transportation of animals (Fletch,
1978; Rowsell, 1990).

The principles enunciated in the CCAC-produced training program continue to be

applicable today. It notes, for example, that personnel engaged in the transportation of
animals, including those employed by the animal facility, require knowledge of the
various types of animals, the differences between species of birds and mammals, as well
as consideration of the invertebrates. They must also be cognizant of the container
requirements, and specification requirements for labelling and marking, and for
completion of the proper documentation, which includes any licensing requirements, both
for export and the country of designation. They should be knowledgeable of shippers'

181
responsibilities as well as those of the individuals receiving the animals. They should
realize the importance of making advance arrangements concerning the shipment and
transport of animals.

The responsibility for knowing about the safe and humane transport of animals includes,
as well, those who, in the conduct of their duties, are exposed to the animals or have
responsibility for the animals they may be carrying, i.e., in the truck, on-board ship or in
the aircraft. The institution receiving the animals should be prepared for accepting the
animals by providing proper facilities and appropriate handling by trained, experienced
personnel.

d) Animal Handling

The animal in a laboratory setting becomes conditioned to the environmental conditions

such as temperature, humidity, air changes, noises, the habits of those working within the
animal room, and animal or human pheremones (substances secreted and released by
animals for detection and response by another of the same species). All of the foregoing
may change when the animals are transported to the facility or moved within it (Slatnetz,
Fratta, Crouse et al. 1957; Baker, Lindsey and Weisbroth, 1979; Gibson, Paterson and
Conville, 1986; Aguila, Pakes, Lai et al. 1988; Rowsell, 1988). Unfortunately, many of
these changes have not been measured precisely (Yousef, 1988). As well, questions have
been raised as to the suitability of certain tests used to measure the stress induced by
environmental changes. Behavioural changes such as increased agitation, difficulty in
handling, reluctance to eat or to drink, bristling of the coat, hiding, and abnormalities in
behaviour may go largely unnoticed, but may cause variations in experimental data.
Therefore, it is essential that the animal be allowed to equilibrate to a new environment.
Because the period for equilibration of each animal and between species varies,
knowledge of the species and of the individual animals is essential.

When the animal arrives at its destination, the institution must ensure that it is then
brought to the institution in a safe and humane manner. Air conditioned vehicles
specifically designed for this purpose are essential in order to reduce stressors that may
have increased during the transport period. Acclimatization to the environment and a
stabilization of the animal, physiologically and behaviourally, are essential prerequisites
before the animal is used. Landi, Kreider, Lang et al. (1982) demonstrated that, following
air transportation of rodents, a two-week period was required for blood and stressor
parameters to return to normal. The adherence to the principles of humane transportation
and handling throughout the transport period and on arrival at the institution should help
ensure that, when animals are used in research, teaching, or testing, the results are
meaningful and scientifically valid.

3. Breeding

A review of modern methods of breeding laboratory animals is beyond the scope of this
Guide. However, it is axiomatic that all types of animals involved in a breeding program
for animal production research purposes will require the best of care and that accurate

182
breeding records must be kept (Box, 1976). It is essential that breeders and researchers
who propose to breed animals for specific research purposes acquire detailed information
on the anatomy, behaviour and physiology of reproduction relating to the stocks
concerned (Altman and Dittmer, 1972; Greep, 1974; Hafez, 1970; Crawford, 1990).
Blaffer-Hrdy and Whitten (1987) present comparative data for non-human primates
(NHP) on cycle length, duration of menstrual flow, visual signals, and the behaviour of
males as well as females in estrus, for all species. Breeders should refer to the sections of
this Guide dealing with management, caging and special housing of the various species.

If an animal model of human disease is to be produced, it is mandatory that the scientist

or breeder fully appreciates the basic physiological and pathological processes involved
in the defect. The research facility is generally the best location for this type of breeding
program. However, under some circumstances it may be preferable to contract for supply
of a particular mutant strain, either with a reliable commercial enterprise or by
arrangement with some other scientist who is already breeding and utilizing the same
model (ILAR, 1979). The breeding of any laboratory animal must be done in accordance
with the accepted genetic standards and genetic nomenclature (Festing, Kondo, Poiley et
al. 1972; ILAR, 1979; Lyon, 1981; Lyon and Searle, 1989), subjects which are beyond
the scope of this Guide.

The decision to establish a breeding colony program in a research institution is one that
the investigator and the institutional ACC should always study carefully in terms of the
nature of the project. Unless breeding is an integral, even essential, part of the research or
teaching exercise, thorough evaluation ought always to be undertaken of: a) the ultimate
real cost (to the institution) of animals bred within the research facility; b) the occupancy
of valuable and costly space which will no longer be available for other research; and c)
the ultimate numbers of animals that will have to be produced versus the actual numbers
that will be utilized. Small in-house breeding programs almost always involve the need to
dispose of animals superfluous to the needs of the project, and the maintenance of excess
breeders in order to try to cope with fluctuating demands.

In commercial livestock production, selection may be based on ancestry, individual

performance or progeny or combinations of these factors. However, in small laboratory
species, emphasis has been placed on maintaining genetic purity (Festing, Kondo, Poiley
et al. 1972). In recent years, questions raised about the genetic homogeneity of some
strains has led to the genetic testing of animals. The larger suppliers of small laboratory
animals provide genetic control for their stocks and strains, and may provide genetic
testing services to investigators using other strains.

The genetic nature of an animal population can be changed in three ways: by selection,
through manipulation of the breeding system, or by altering the genome through the
introduction of alien genes (DeTolla, 1991). Traditionally, the rapidity with which genetic
change in a population is achieved will depend in part on the sex of the selected animals;
the female has less effect on the maximal selection differential than the male, as the latter
can produce many more progeny. The decision as to which animals are to be bred will
depend on numerous criteria directly related to the purpose of the breeding exercise.

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4. Breeding Transgenic Animals

The time required to produce large numbers of transgenic animals will depend on the
reproductive capacity of the animal. Transgenic animals are now considered a standard
biomedical research tool (Saffer, 1992) and are increasingly being used as animal models
for human diseases (Merlino, 1991), gene therapy, the study of virus-induced disease, the
physiology of foreign gene expression (Palmiter and Brinster, 1986; Geistfeld, 1991),
probes into complex systems (Hanahan, 1989), and as models for genetic toxicology
studies (Myhr and Brusick, 1991). Their use is increasing, and indeed, has been cited as a
major cause in the first rise in animal use in the U.K. for many years (Anon., 1992).

Management of a transgenic mouse colony has recently been described by Geistfeld

(1991), and a procedures manual has been published by Hogan, Constantini and Lacey
(1986). As well, there are numerous research reports (Brinster, Chen, Trumbauer et al.
1985; Bishop and Smith, 1989; Depamphilis, Herman, Martinez-salas et al. 1988;
Gordon, Scangos, Plotkin et al. 1980; Jaenisch, 1976, 1988) describing transgenic animal
use. Britain's Health and Safety Executive published Guidelines on Work with Transgenic
Animals (1989) (Baynards House, 1 Chapstow Place, London, W2) which must be
followed when creating, breeding or handling transgenic animals in the U.K. (Connor,
1989).

In describing management of a transgenic mouse colony Geistfeld (1991) suggests that

Caesarian-derived pups be used in order to eliminate pathogens. Various breeding
systems are feasible; usually a 2:1 to 3:1 harem mating system is effective, although there
are no hard and fast rules.

Some of the problems associated with the breeding of transgenic mice have been outlined
by Donnelly and Walsh-Mullen (1991). These include contamination of the media used in
collecting eggs and blastocysts for microinjection, because of which the surrogate dam
fails to deliver. They note that introducing foreign genes may cause deleterious insertions
that prove lethal to the animal or that compromise reproduction.

The CCAC recently established a Committee on Animal Biotechnology (CABT), which

has as its mandate: "to develop ongoing guidelines for embryo manipulation, fetal
research, and transgenic animals". The CABT considers it acceptable for
transgenic/embryo manipulation research to produce animals which do not have a
negative impact on the animal's well-being or on the environment, and which have a
positive, scientifically justifiable endpoint. Should transgenic technology result in a new
species or strain of animal, research techniques must be developed to adequately test the
impact of such animals, in the view of the Committee. Some of Canada's institutions have
prepared their own guidelines for research involving transgenics.

5. Animal Models with Special Needs

Animal models of human disease are used to study the causes and therapeutic and

184
preventive methods for human disease, as well as to develop new drugs (Nomura,
Katsuki, Yokoyama et al. 1987). Models are available for many diseases and conditions,
e.g., hemophilia (Moake, 1988) atherosclerosis (Reddick, Read, Brinkhous et al. 1990;
Farrell, Saunders, Freeman et al. 1986), Pasteurellosis (Morck, Costerton, Bolingbroke et
al. 1990), intestinal disease (Pfeiffer, 1985), hepatic degeneration (Hultgren, Stevens and
Hardy, 1986), enteric diseases such as Campylobacter jejuni (Fox, Ackerman, Taylor et
al. 1987), cardiomyopathy (Wagner, Reynolds, Weisman et al. 1986) and neurological
disease (Barnes, 1986). Animal models for advancing understanding of diseases such as
hypertension, gastrointestinal tract and cardiovascular disease were discussed at a
symposium of the British Laboratory Animals Veterinary Association (BLAVA) (Anon.,
1986). Another meeting addressed the challenges facing researchers into the human
immunodeficiency virus (HIV) and the need for animal models in this regard (Groopman,
1991).

Animal models of some conditions or diseases have special needs beyond those of
normal, healthy laboratory animals. These special needs must be recognized and
accommodated when such animal models are going to be used in research. It should be
the responsibility of the principal investigator to take into consideration the special needs
of the animals before embarking on the research project. These special needs will no
doubt impact on the research budget in terms of additional animal care time, materials,
and equipment. ACC reviews of research proposals should include an assessment of these
extra considerations for the animals.

The principle that encompasses this responsibility to attend to the special needs of animal
models could be stated as follows: that any pain, suffering, distress, or deficits in function
that negatively affect the animal's well-being, not scientifically "necessary" for the study,
should be alleviated or minimized. Cost or convenience should not deter from this.
Further, as soon as the study is done, the animal suffering should be terminated (Olfert,
1992).

6. Identification of the Sexes

Generally, sexes are kept separated subsequent to weaning, except for breeding purposes
or because of experimental design requirements. Unwanted matings amongst stock
animals should not occur, and under experimental conditions may compromise the
experimental results.

The sexing of animals may be difficult in the newborn or in species with which one is
unfamiliar. In addition to the genital organs themselves, secondary sex characteristics can
be used in some cases (Valle, 1990). Detailed descriptions of the techniques/observations
used to sex the various common laboratory animal species can be found in the species
chapters of the CCAC Guide Volume 2, or in other general books on species used as
laboratory animals (Poole, 1987). Descriptions of the techniques used to determine the
sex of some of the less common species are also available (Goin and Goin, 1971; Frye,
1991; Marcus, 1981).

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B. RESTRAINT AND MANIPULATIONS

1. Physical Restraint

a) Introduction

It is necessary to restrain most animals for even the simplest of procedures (e.g., to take
the animal's temperature). When short-term chemical restraint (e.g., anesthetic,
tranquillizer, etc.) is not possible and/or is not compatible with the experimental
requirements, some form of physical/mechanical restraint may have to be used.

It is well known that the quality of the restraint will influence the animal's response
(Hemsworth, Barnett and Hansen, 1986). With laboratory rodents, Lee (1992) contends
"there is no superior restraint than an able technician aide's hands." It has been shown that
an experienced technician picking up a rat disturbs the animal less than a totally
inexperienced technician who is afraid of being bitten (Barclay, Herbert and Poole,
1988). Immobilization stress is known to have an effect on rat performance (Grilly and
Gowans, 1986). Par and Glavin (1986) have reviewed restraint-induced stress.

Animals become accustomed to repeated, gentle, short-term restraint which is not

accompanied by stressful or painful procedures. Longer-term restraint is accompanied by
evidence of stress unless the animal has undergone a prolonged period of acclimatization
to the experimental situation, and is not subjected to any painful procedures while it is
being restrained (Golub and Anderson, 1986; Rushen, 1986). The level of distress varies
from animal to animal in the same situation, and may affect experimental results.

Two factors which have a strong influence on the degree of stress experienced by a
restrained animal are the isolation from conspecifics (during restraint), and the degree of
immobilization. These two factors should be considered if the experimental procedure
requires that the animal be restrained. Visual, auditory and olfactory contact with
conspecifics may suffice to reduce stress levels. Sheep, for example, should not be out of
sight and sound of other sheep, even if they are able to move around in the isolation pen
(see also Social and Behavioural Requirements of Experimental Animals).

In all situations in which prolonged physical restraint is required, the use of proposed
restraining methods must first have been justified and received approval through peer
review and the institutional ACC, using the guidelines established by the CCAC. Such
reviews should provide assurance that the technology is either lacking or its use is not
warranted for obtaining the required measurements in the unrestrained animal. The
restrained animal will always require special consideration, care and surveillance (see
also Social and Behavioural Requirements of Experimental Animals; Categories of
Invasiveness statement; Ethics of Animal Investigation document, found elsewhere in this
Guide).

b) General Guidelines for Care of Restrained Animals

186
i) Restraint procedures should only be invoked after all other less stressful procedures
have been rejected as alternatives.

ii) Supervision of animals in restraining devices should only be assigned to fully qualified
and experienced personnel.

iii) The principal investigator has the responsibility to ensure that all members of the
research team, particularly those responsible for day-to-day animal care, are fully aware
of the rationale for the restraint procedures and for the complications for the animal
which may occur as a result of the restraint.

iv) Consultation should be sought with those experienced in the restraint procedures to be
invoked, prior to its initial use, to ensure that minimal restraint is used to accomplish the
experimental goals.

v) Physiologic, biochemical and hormonal changes occur in any restrained animal

(Grtner, Bttner, Dhler et al. 1980; Toth and January, 1990; Moberg, 1992; Bush,
Custer, Smeller et al. 1977; Mayer and Bowman, 1972; Markowitz and Spinelli, 1986),
and investigators should consider how these effects will influence their proposed
experiments.

c) Special Surveillance

The following comments are generally applicable to all restrained animals; however, they
refer especially to restraint in non-human primates (NHP):

i) Regardless of duration of restraint, special attention must constantly be paid to the

possible development of associated ill effects.

ii) A minimum of twice daily, careful physical inspection of each restrained animal is
mandatory.

iii) Inspection should include not only physical examination, but also evaluation of
general behaviour. Food and water consumption, as well as the animal's weight if
possible should be recorded daily. When any unusual manifestations develop, or
physiological parameters show undue variance, immediate therapeutic measures must be
taken.

d) Restraint Devices

Devices developed for short-term laboratory animal restraint include a plastic cage for
restraint of opossum (Thomason and Russell, 1986), an acute restraint device for rhesus
monkeys described as "a practical and inexpensive alternative to the standard primate
chair" (Robbins, Zwick, Leedy et al. 1986), and a full-body retraining device for small
animals that permits the short-term recording of physiologic data (Yagiela and Bilger,

187
1986). The use of slings, pioneered for miniature swine by Panepinto (Panepinto,
Phillips, Norden et al. 1983) has now been extended to rats and rabbits (Kumar, Wong,
Johnson et al. 1979) and other species. Commercial sources of slings for rats and rabbits
are available (e.g., Harvard Bioscience, Ealing Scientific Ltd., 6010 Vanden Abeele St.,
St. Laurent, Quebec Canada H4S 1R9), and for livestock species and dogs (Munk's
Livestock Sling Mfg. Inc., 1143 W. Marches Pt. Rd., Anacortes, WA 98221 USA).

Efforts are being made to provide less restrictive restraint systems that facilitate
movement and permit sampling or protect the instrumentation or equipment (Houghton,
1985; Anderson and Houghton, 1983; Dalton, 1985; Munson, 1974). Examples include;
an intravenous catheter system for long-term (at least six to eight weeks) parenteral
nutrition of unrestrained rats (Brenner, Muller, Walter et al. 1985), a backpack system for
mini-pump infusions in marmosets (Ruiz de Elvira and Abbott, 1986), and a tethering
system for intravenous and intragastric drug administration in the baboon (Lukas,
Griffiths, Bradford et al. 1982).

Scientists are continuously seeking new methods which will lessen the stress involved in
the collection of samples such as bile (Rath and Hutchison, 1989; Kanz, Vanoye-Trevino
and Molsen, 1989), and blood (Lawhorn, 1988). It should be re-emphasized that efforts to
reduce the stress of monitoring often involve the training and socialization of the animal
to the procedure (Vanderlip, Vanderlip and Myles, 1985a, 1985b).

The advent of vascular access ports has made it possible to take repeated blood samples
from, or administer drugs to, a wide variety of animals with the minimum of restraint
(Houghton, 1985). These ports consist of an intravascular catheter attached to a reservoir
with a diaphragm (Dalton, 1985; Harvey-Clark, 1990). The appropriate blood vessel is
cannulated and the reservoir is located subcutaneously at a suitable location. With the use
of a topical, penetrating analgesic cream, access to the port can be achieved painlessly.
Samples must be taken using sterile technique, in order to avoid contamination of the
vascular access port. These ports are usually maintained patent by filling them with a
heparin solution, and may remain functional for many months or years.

Another example of a less restrictive restraint system that facilitates animal movement,
yet permits sampling or protection of instrumentation or equipment is the "Jacket and
Swivel Tethering System" developed by Chatham (1985). It allows much greater
movement and permit simultaneous fluid sampling, drug infusion, and electronic
measuring and monitoring. The tethered animal can be maintained comfortably and
humanely for months, while fully instrumented. The light weight and flexibility of the
tether, combined with the low friction of the swivel, inhibits very little of the animal's
normal cage activity. (Alice King Chatham Medical Arts, 5043 Oaknoll Ave., Los
Angeles, CA 90043; Harvard Bioscience, Ealing Scientific Ltd., 6010 Vanden Abeele St.,
St. Laurent, Quebec Canada H4S 1R9.)

The "Pole and Collar" system (Anderson and Houghton, 1983) provides a standardizable,
efficient means of training monkeys to calmly leave their cages, enter restraint devices
for as long as the procedures require, and return to their cages without the need for

188
chemical restraint or forced manual submission (Houghton, 1985).

Biotelemetry systems are increasingly being used to transmit biological information from
animals to a remote location. A variety of biotelemetry systems are available, from
backpack systems to those that are totally implantable (Halpryn, 1985).

2. Implantation, Cannulation and Sampling

Chronic studies involving the implantation of electrodes, cannulae and catheters will
require that the animal be anesthetized at the time of implantation, and restrained awake
when sampling is undertaken.

Electrode implantations must be undertaken by, or under the direct supervision of

personnel experienced in the techniques involved, and must utilize proper surgical and
anesthetic procedures (Meyer and Meyer, 1971; NIH, 1991) (see also Standards for
Experimental Surgery and Anesthesia). Detailed descriptions of stereotaxic surgical
techniques may be found in several sources (Hart, 1969; Pellegrino and Cushman, 1971;
Skinner, 1971; Singh and Avery, 1975).

3. Bleeding

Guidelines for blood removal from laboratory mammals and birds have recently been
published in Great Britain (BVA/FRAME/RSPCA/UFAW Joint Working Group on
Refinement, 1993). Efforts should constantly be made to refine scientific techniques so as
to reduce the volume of the blood sample. In small animals such as mice, volume and
frequency are of particular importance. If the animal's welfare is threatened by the
volume of the sample required, either more animals should be used, or compensatory
blood transfusion considered.

Rather than multiple sampling carried out by repeated needle punctures, a butterfly
needle or a percutaneous (over the needle) cannula taped in position, should be utilized.

In removing volumes greater than 0.1 ml, as large a bore as possible should be used in
order to ensure rapid blood withdrawal without collapsing the vein, with the constraint of
avoiding hematomata formation. Before taking a sample, it is important to accurately
locate the vein and dilate it by gentle obstruction or warming. If general body warming is
used, the animal must be constantly observed to prevent hyperthermia, as evidenced by
more rapid breathing, panting or salivating. The use of xylene (xylol, dimethylbenzene)
as a dilator is not recommended as it causes skin rashes and is easily misused.

Common bleeding sites may be found in Appendix VIII of this Guide.

4. Motivation Procedures

In the conduct of behavioural studies, the use of positive reinforcement (e.g., reward in
the form of a preferred food) is preferable to use of aversive stimulation (Lea, 1979).

189
CCAC's Ethics of Animal Investigation (found elsewhere in this Guide) also notes that
investigators, ACCs and Review Committees are advised to "be especially cautious in
evaluating...electric shock as negative reinforcement." Elsewhere in this Guide under the
Use of Animals in Psychology, it is noted:

"Similarly, when researchers use electric shock as a means of producing stress or

motivating animals to escape or avoid, they are fully recognizant that electric shock does
not normally occur in nature. They do assume, however, that this particular, easily
controlled aversive event can serve as a model for analogue of other unpleasant events
that do occur naturally and affect behaviour...."

Experimenters are urged to use the least aversive shock intensity x duration x frequency
combination that is compatible with the goals of the research (Olfert, 1992). Particular
values of these parameters will thus vary with species, and with the goal of the research.
In many instances, there are well-developed procedures for determining the appropriate
value of shock, based upon behavioural criteria. For example, "titration" procedures
allow the minimum value of shock that will maintain a given behaviour to be determined
for each individual animal. In all cases, it behooves the investigator to base the selection
of shock values upon behavioural criteria, so as to use the least aversive shock that will
permit collection of orderly data and the success of the research. For example, the
arbitrary selection of a "low", a "medium" and a "high" intensity, without careful
consideration of the behavioural effects of these values and the interaction of the elicited
behaviours with the desired behavioural outcomes, may cause undue suffering and waste
of research animals, and affect the research.

When shock is being used in combination with other stressors, the investigator should be
aware of the possibility of summation of negative effects.

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Chapter VIII - Occupational Health and Safety

VIII. OCCUPATIONAL HEALTH AND SAFETY

Those working with experimental animals risk exposure to physical hazards (e.g., heat,
noise, radiation), chemical hazards (e.g., disinfectants, cleaning solutions), as well as
intestinal parasites, enteric bacteria, pathogenic organisms, and animal bites (Soave and
Brand, 1991). As well, those working with swine in confinement buildings may suffer
future chronic and irreversible lung damage, according to Donham and Leininger (1984).
Those working with non-human primates (NHP) must take special precautions. Guidance
for these individuals may be found in Volume 2 of this Guide.

A. REGULATORY REQUIREMENTS

As is the case with other laboratories, the animal care facility should have an
Occupational Health and Safety program. All persons using the facility should also be
familiar with the requirements of relevant federal, provincial and municipal legislation.
This would include, for example, the federal Health of Animals Act (38-39 Elizabeth II,
Chapter 21, pgs. 387-421), which replaced the Animal Disease and Protection Act and
which governs control of animal diseases and toxic substances. Those working with

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animals should also be cognizant of institutional and/or facility safety program (see also
Volume 2 of this Guide [CCAC, 1984a]).

The Workplace Hazardous Materials Information System (WHMIS), which resulted from
federal and provincial co-operation, was instituted in 1988. Federal government
laboratories are governed by federal WHMIS and the Canada Labour Code. The
following publications are available free of charge from Labour Canada: The Employer
and WHMIS; Introduction to the WHMIS Program; Exercise WHMIS in the Workplace,
and a relevant poster.

Elsewhere, provincially enacted Health and Safety legislation specifies the accountability
of owners and directors and the rights and responsibilities of employers, supervisors and
workers in the workplace. The right to refuse unsafe work is a part of the Occupational
Health and Safety (OHS) Act. WHMIS regulations are also a section of this legislation
and require that each employer provide safe working conditions and that employees be
informed about all hazards they will face in the course of their duties. Employees are also
given the right to withdraw from the workplace if faced with an unsafe condition. All
hazardous substances, including microorganisms, must be labelled in a specified manner,
and a Material Safety Data Sheet (MSDS) must be available to accompany each
hazardous substance. Each province has adapted these federal government guidelines for
its own purposes. WHMIS material may be obtained from provincial Ministries of
Labour.

All personnel working with animals must understand how to handle the species involved,
both for their own safety and health, and for that of the animals. Training for this should
be provided.

B. BIOLOGICAL HAZARDS

Guidelines for working with biohazards (e.g., bacteria, viruses, parasites, fungi and other
infectious agents), are provided in the Health and Welfare Canada/Medical Research
Council Laboratory Biosafety Guidelines (HWC/MRC, 1990). The guidelines include
such items as biohazard containment, laboratory design, personal hygiene and safety
facilities, and can be used to provide training for employees as mandated by WHMIS.

The biosafety guidelines apply to all research carried out or supported by the federal
government and have been adopted by many industries.

Standard Operating Procedures (SOP) based on the guidelines, aimed at minimizing risks
to humans in biohazard risk areas, should be developed and enforced.

Personal cleanliness is an important barrier to infection and washing of hands after

handling any animal will reduce the risk of disease spread and self-infection. All
employees working with animals, as well as visitors to the facility, should wear protective
clothing, minimally a lab coat.

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All contaminated material must be decontaminated before disposal. Necropsy of animals
infected with highly infectious agents should be carried out in certified and tested
biological safety cabinets. Necropsy material for disposal should be sealed in plastic
bags, properly labelled and incinerated. The necropsy room should be properly equipped
to provide adequate refrigeration and hand-washing facilities.

C. ZOONOSES

Those infections that are "secondarily transmitted from animals to humans" are referred
to as zoonoses (Schnurrenberger and Hubbert, 1981; August and Loar, 1987; Acha and
Szyfres, 1989) and can seriously affect research (Hamm, 1986; Bhatt, Jacoby, Morse et
al. 1986; ILAR/NRC, 1991).

While most infectious agents show a considerable degree of species specificity, they also
may, from time to time, vary widely in virulence and in their capacity to break through
species barriers. Thus, infections that have not commonly been considered to be zoonotic
hazards may sporadically affect susceptible persons or animals. Persons potentially at
higher risk are those who suffer from defective immune systems and those who are under
severe stress or who have non-overt clinical disease. Numerous pathogenic
microorganisms, such as those responsible for tuberculosis, brucellosis, rabies, etc.,
which are normally perpetuated by direct transmission from one or more species of
vertebrate animals, are also readily transmissible to humans.

Transmission of infections from animals to humans can generally be avoided through

proper veterinary care and adherence to SOPs for control of transmission. However,
when animals are obtained from areas in which zoonotic diseases are known to exist, e.g.,
in NHP acquired from the wild (Houghton, 1986) special attention is required.

Work involving exposure to hazardous microorganisms might require prior immunization

of the staff, if a vaccine is available. It is recommended, for example, that all personnel
handling random-source dogs and cats, including dealers and handlers, should receive
routine rabies vaccination (see also Special Animal Colonies, Infectious Disease Units).

Serological testing and banking of reference serum samples from all personnel working
in the animal facility is advisable. This is of particular importance where NHP are being
handled and/or agents infectious to humans are being used.

Caution should be exercised in assigning women of childbearing status to animal care

duties that might expose them to potential or known teratogens. For example Toxoplasma
gondii, a protozoan that infects most species of warm-blooded animals, including
humans, is spread primarily by oocysts shed in cat feces. These oocysts sporulate in two
to four days and may survive for more than a year (Fraser and Mays, 1986). Human
toxoplasmosis can result in spontaneous abortion, prematurity, stillbirth or congenital
defects (Schnurrenberger and Hubbert, 1981).

The life cycle of the causative organisms implicated in a number of indirect zoonoses

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may involve transmission through one or more other vertebrate and/or invertebrate
intermediate hosts before affecting humans (for example, in taeniasis, tularemia, and
vesicular stomatitis). Amongst invertebrate vectors of zoonotic disease, the biting insects
are the main offenders. A list of some of the diseases transmitted to humans from animals
is included in Appendix VII.

The role of cold-blooded vertebrates in the epidemiology of zoonoses should not be

overlooked. In particular, turtles infected with salmonella may constitute a human health
hazard in the student laboratory as well as in the animal facility (Sherris, 1990).

D. PROCEDURES FOR WORKING WITH NON-HUMAN PRIMATES

This topic has also been discussed in Volume 2 of this Guide (CCAC, 1984b).

All animals must be regarded as potential sources of zoonoses, although the risk of this
occurring will vary widely with the class, species, and source of the animal involved. In
general, the more closely related phylogenetically a species is to humans, (Anon., 1987a,
1987b; FRAME, 1987; Rice, 1987a, 1987b), the greater the likelihood of zoonoses. It is
for this reason that special precautions must be followed for NHP (Love, 1980; Wong and
Gardell, 1982; Richter, Lehner and Henrickson, 1984; Else, 1988).

Each institution that maintains a NHP facility is responsible for providing the proper
veterinary and human medical services to safeguard the health and safety of both
personnel and animals. International guidelines have been prepared for those working
with NHP (FRAME/CREA, 1987; Kaplan, 1987; Anon., 1989; MRC, 1985).

Outbreaks of viral diseases, e.g., Callitrichid Hepatitis Virus (Anderson, 1991) recently
"rocked the primatologist's world," and current procedures for the rapid diagnosis of
primate viral diseases include serology, virus isolation, direct visualization using
electronmicroscopy or immunofluorescence, and detection of viral components (Kalter
and Herberling, 1990).

The most reasonable and effective approach in reducing occupational infection risks is to
develop and follow SOPs that preclude or minimize overt occupational exposure among
personnel working with NHP or biological samples therefrom. SOPs should be
established for an Occupational Health and Safety Program for personnel which would
include serological screening and vaccination, use of protective clothing, containment,
stress on personal hygiene, procedures for accidents including bite wounds and/or other
exposure to potential risk, and for quarantine and quality control procedures for confined
animals.

Guidelines have been prepared for prevention of Herpes Simiae (B-Virus) infection by a
B-Virus Working Group which convened at the Centers for Disease Control (Anon.,
1987c; Kaplan, Balk, Brock et al. 1987; Schulhof, 1990). Herpes Simiae is fatal in
humans (Kalter and Herberling, 1989).

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Similarly, because of the expanding use of Simian Immunodeficiency Virus (SIV), which
is closely related to the Human Immunodeficiency Virus (HIV), guidelines in this regard
have been prepared as well (Anon., 1989). Standard serological procedures to identify
SIV antibody are used in laboratories conducting research with the virus, and the
National Institutes of Health (NIH) and World Health Organization (WHO) have
expanded their diagnostic services (Kalter, 1987).

In addition to NHP from the wild, those within the colony may also carry indigenous
latent infections (Baulu, Everard and Everard, 1987; Dance, King, Aucken et al. 1992). It
is important to establish rigid quarantine and quality control procedures for the animal
colony and to better define and be more aware of the potential risks. The Ebola-like virus
outbreak in the U.S. in 1989 exemplified such risks (Anon., 1990; Anderson, 1990a,
1990b; Dalgard, Hardy, Pearson et al. 1992).

Adherence to the following list of precautions is recommended:

a) all NHP must be considered as potentially carrying a disease transmissible to humans;

b) NHP, or anything that has been in direct contact with them, should not come in contact
with the skin;

c) protective clothing, including coveralls, boot covers, surgical caps, masks and gloves
should be worn when working with NHP, and removed when leaving the NHP quarters;

d) smoking and bringing food and drink into NHP rooms are strictly forbidden;

e) facilities for washing hands must be made available and used by all personnel
immediately upon leaving NHP rooms;

f) personnel with sores, cuts, and other lacerations should not come in contact with NHP.
However, if this is unavoidable, then the lesion must be adequately covered prior to and
during any activity in a room containing NHP, and dressings must be changed
immediately upon leaving. These dressings and any other disposable items so exposed
must be treated as biological hazardous waste;

g) all cuts, bites, scratches, or needle punctures acquired while working with or in
proximity of NHP must be reported to the medical authority designated by the institution.
SOPs for all wounds so encountered should be developed and followed accordingly.
Immediate treatment must ensure that the wound is made to bleed freely and thoroughly
scrubbed and cleansed with soap and water. A flushing of the wound area with a
provodine iodine solution is recommended. In the event that sterility has been breached
(e.g., tearing or puncture of a surgical glove) the hands must be re-scrubbed before
leaving the room and re-gloved before continuing the procedure.

Following injury by a NHP, the animal concerned must immediately be immobilized and
examined for excessive salivation and for lesions of the oral cavity which may be

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characteristic of Herpes (B-Virus). SOPs must be followed for dealing with this type of
accident. Procedures for sampling for Herpes B of the animal and of the injured person
must be followed. The results of the examination must be communicated to the
previously designated medical authorities, along with information on the species of NHP,
length of time in the colony, and contacts with other species;

h) special precautions should be taken whenever conducting necropsies on NHP that have
died during the conditioning period; necropsy procedures should include the wearing of
protective clothing, surgical caps, and masks, gowns and surgical gloves. The use of
biosafety cabinets for conducting all necropsy of NHP tissue is
recommended;

i) because of the possible danger of contacting Hepatitis A, it is recommended that

personnel working with newly imported chimpanzees receive hyper-immune serum
globulin prophylactically. Animals should be tested for human hepatitis antigens and, if
positive, strictly quarantined;

j) all personnel having contact with NHP must be free of tuberculosis and should receive
a tuberculin skin test not less than once yearly and X-ray examination as prescribed. It
should be noted that it has recently been reported that misleading positive tuberculin
reactions were caused in squirrel monkeys that had received Freund's Complete Adjuvant
(FCA) (Pierce and Dukelow, 1988);

k) protective leather gauntlets should be worn when handling conscious NHP. Several
varieties are available commercially;

l) all laundry that has been in direct contact with NHP or their excreta should be
autoclaved prior to being sent out for washing.

E. ALLERGIES

Allergies to laboratory animals are a significant occupational health concern for people
regularly working with the common laboratory animal species (Aoyama, Ueda, Manda et
al. 1992; Olson, 1986; Bland, Levine, Wilson et al. 1986; Botham, Davies and Teasdale,
1987; Kibby, Powell and Cromer, 1989; Lutsky, 1987; Slovak and Hill, 1987; Venables,
Tee, Hawkings et al. 1988). Laboratory animal allergy (LAA) is an immediate-type
hypersensitivity reaction, IgE-mediated, which develops upon exposure to a laboratory
animal, its fur or dander, its urine, saliva, serum or other body tissues. Typical symptoms
range from mild (e.g., upper respiratory signs such as sneezing, itchy and/or runny nose
and eyes, and skin reactions such as red, raised and itchy wheals after contact with
animals, their tissues or their excreta), to severe [e.g., wheezing, shortness of breath, and
a feeling of chest tightness (asthma)]. Persons experiencing such symptoms should be
advised to contact their physician for diagnosis and treatment.

Measures which can reduce the degree of exposure to laboratory animal allergens
include:

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a) use of protective gear such as gloves, face masks, gowns, shoe covers, etc., worn only
in animal rooms;

b) regular hand-washing, and showering after work;

c) use of improved filtration in animal room ventilation systems, and the use of special
filtered caging systems; and

d) educational programs for employees identifying high risk (e.g., high allergen load)
areas and tasks, and strict use of preventive measures, as set out by the institution's SOPs.

Institutions are encouraged to include a consideration of LAA in their Occupational

Health and Safety programs. As noted above, identifying high risk areas and tasks
(Eggleston, Newill, Ansari et al. 1989; Gordon, Tee, Lowson et al. 1992; Swanson,
Campbell, O'Hallaren et al. 1990), and the use of SOPs in these areas, along with
education of staff, are useful in reducing the severity of the problem (Botham, Davies and
Teasdale, 1987). Procedures for monitoring exposure, health-monitoring of staff at risk,
and for dealing with staff who become allergic should also be considered (Botham,
Davies and Teasdale, 1987; Lutsky, 1987; Newill, Evans and Khoury, 1986).

F. PHYSICAL INJURIES AND CHEMICAL HAZARDS

Physical injuries related to the handling of animals may be kept to a minimum by

ensuring that:

a) all staff are trained and experienced in handling the species with which they work, and
that they know the particular hazards associated with each species;

b) all staff are familiar with the hazards of the experiment, and are provided with (and
use) a proper working area, protective clothing and equipment;

c) a mechanism is in place in every unit to deal with animal-inflicted injury, and for
referral for any further medical treatment if this is required.

Responsibility for ensuring that first aid kit(s) are available and always properly stocked
must be clearly identified. The location of the first aid kit(s) should be prominently
marked and all personnel using the facility should be made aware of these locations.

Injuries from chemicals can be avoided by treating all chemicals with care, by knowing
their properties and adhering to the accepted safety practices for handling that type of
product. WHMIS, legislative and institutional requirements must be met.

Care should always be taken in handling such common chemicals as industrial detergents
used in cage washers, cleaning agents, and powerful disinfectants. These substances

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should be stored separate from animal feed and bedding materials. Volatile liquids used
as anesthetics or for euthanasia, and other toxic and volatile materials, should be stored in
well-ventilated fume hoods or cabinets designed for that purpose.

G. RADIATION AND ULTRAVIOLET LIGHT

Radioactive materials present special hazards. All persons working with these materials
should know the properties of each, and be familiar with the appropriate safe handling
techniques. The possession of radioactive materials is authorized by Radioisotope
Licences issued by the (federal) Atomic Energy Control Board (AECB) to the
institutions. The Radiation Safety Program is administered by a Radiation Safety Officer,
who the AECB recommends sit as an ex-officio member of the institution's Occupational
Health and Safety Committee. Use of X-rays is governed by Occupational Health and
Safety Acts under provincial Ministries of Labour.

Isotope-treated animals may pass radioactive material in their excrement, which should
therefore be disposed of in an approved manner, as must the animal itself after death.
Complete records should be kept through to the final disposition of these animals.

The eye and skin are critical areas for exposure to ultraviolet (UV) light. The eye, in
particular, can be seriously injured. Staff should not be exposed to UV rays; however, if
they must be, they should be warned of the hazards and provided with "wraparound"
safety glasses. As well, the source of illumination should be suitably marked. The
maximum intensities tolerated by sensitive faces for a seven-hour day, range from 0.1 to
0.5 milliwatt per square foot.

H. REFERENCES

ACHA, P.N. and SZYFRES, B. Zoonoses and communicable diseases common to man
and animals. 2nd Ed. Washington, DC: Pan American Health Organization/World Health
Organization, 1989.

ANDERSON, G.C. Monkey imports may be curtailed in U.S.. Nature March 22, 1990a:
280.

ANDERSON. G.C. U.S. shuts down monkey trade. Nature March 29, 1990b: 369.

ANDERSON, G.C. Emerging virus threat. Nature May 9, 1991: 89.

ANON. My close cousin the chimpanzee. Science, Research News 1987a; 238: 273-275.

ANON. B-Virus infection in humans. Lab. Primate Newsl. 1987b; 26(3): 2-4.

ANON. The use of non-human primates in laboratories. The Lancet Jan. 31, 1987c: 286.

ANON. Guidelines to prevent Simian Immunodeficiency Virus infection in laboratory

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workers and animal handlers. Lab. Primate Newsl. 1989; 28(1): 17-21.

ANON. Concurrent Ebola and SHF in imported primates. Lab. Primate Newsl. 1990;
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AOYAMA, K. UEDA, A., MANDA, F. et al. Allergy to laboratory animals: an

epidemiological study. Brit. J. Indust. Med. 1992; 49: 41-47.

AUGUST, J.R. and LOAR, A.S. Zoonotic diseases. Vet. Clin. North Amer. Sm. Anim.
Pract. 17(1), 1987. Philadelphia, London, Toronto, Mexico City, Rio de Janeiro, Sydney,
Tokyo, Hong Kong: W.B. Saunders Co.

BAULU, J., EVERARD, C.O.R. and EVERARD, J.D. Leptospires in vervet monkeys
(Cercopithecus aethiops sabaeus) on Barbados. J. Wildl. Dis. 1987; 23(1): 63-68.

BHATT, P.N., JACOBY, R.O., MORSE, H.C. III et al. Viral and mycoplasmal infections
of laboratory rodents. Effects on biomedical research. Orlando, San Diego, New York,
Austin, Boston, London, Sydney, Tokyo, Toronto: Academic Press, 1986.

BLAND, S.M., LEVINE, M.S., WILSON, P.D. et al. Occupational allergy to laboratory
animals: an epidemiological study. J. Occup. Med. 1986; 28(11): 1151-1157.

BOTHAM, P.A., DAVIES, G.E. and TEASDALE, E.L. Allergy to laboratory animals: a
prospective study of its incidence and of the influence of atopy on its development. Brit.
J. Indust. Med. 1987; 44: 627-632.

CANADIAN COUNCIL ON ANIMAL CARE. Guide to the care and use of experimental
animals, Volume 2. Ottawa, Ont.: CCAC, 1984a: 170-172.

CANADIAN COUNCIL ON ANIMAL CARE. Non-human primates. In: Guide to the

care and use of experimental animals. Vol. 2. Ottawa, Ont.: CCAC, 1984b: 163-173.

DALGARD, D.W., HARDY, R.J., PEARSON, S.L., PUCAK, G.J., QUANDER, R.V.,
ZACK, P.M., PETERS, C.J. et al. Combined simian hemorrhagic fever and ebola virus
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DANCE, D.A.B., KING, C., AUCKEN, H., KNOTT, C.D., WEST, P.G. and PITT, T.L.
An outbreak of melioidosis in imported primates in Britain. Vet. Rec. 1992; 130(24): 525-
529.

DONHAM, K.J and LEININGER, J.R. Animal studies of potential chronic lung disease
of workers in swine confinement buildings. Amer. J. Vet. Res. 1984; 45(5): 926-931.

EGGLESTON, P.A., NEWILL, C.A., ANSARI, A.A. et al. Task-related variation in

airborne concentrations of laboratory animal allergens: Studies with rat n1. J. Allergy
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FRASER, C.M. and MAYS, A. The Merck veterinary manual. 6th Ed. Rahway, NJ:
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FUND FOR THE REPLACEMENT OF ANIMALS IN MEDICAL EXPERIMENTS/

COMMITTEE FOR THE REFORM OF ANIMAL EXPERIMENTATION. The use of
non-human primates as laboratory animals in Great Britain. Nottingham: FRAME, 1987.
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from laboratory rats. Brit. J. Indust. Med. 1992; 49: 416-422.

HAMM, T.E. Jr., ed. Complications of viral and mycoplasmal infections in rodents to
toxicology research and testing. Washington, New York, London: Hemisphere Pub.
Corp., 1986.

HEALTH AND WELFARE CANADA/MEDICAL RESEARCH COUNCIL OF

CANADA. Laboratory biosafety guidelines. Cat. No. MR 21-1/1990E. Ottawa, Ont.:
Supply and Services Canada, 1990.

HOUGHTON, P. Collecting feral cynomolgus macaques. Lab Animal 1986; 15(5):

19,20,22.

INSTITUTE OF LABORATORY ANIMAL RESOURCES/NATIONAL RESEARCH

COUNCIL. Infectious diseases of mice and rats. Washington, DC: National Academy
Press, 1991.

KALTER, S.S. Simian AIDS testing available. Lab Primate Newsl. 1987; 26(1): 4.

KALTER, S.S. and HERBERLING, R.L. B-virus infection of primates in perspective.

Lab Animal 1989: 31-34.

KALTER, S.S. and HERBERLING, R.L. Current procedures for the rapid diagnosis of
primate viral diseases. Lab Animal 1990: 39-47.

KAPLAN, J.E., BALK, M., BROCK, B. et al. (correspondence) Guidelines for

prevention of Herpesvirus Simiae (B-virus) infection in monkey handlers. Lab. Anim.
Sci. 1987; 37(6): 709-712.

KIBBY, T., POWELL, G. and CROMER, J. Allergy to laboratory animals: A prospective

and cross-sectional study. J. Occup. Med. 1989; 31(10): 842-846.

LUTSKY, I. A worldwide survey of management practices in laboratory animal allergy.

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MEDICAL RESEARCH COUNCIL. The management of simians in relation to

infectious hazards to staff. Council's Simian Virus Committee, 1985: 1-17.

NEWILL, C.A., EVANS, R. and KHOURY, M.J. Preemployment screening for allergy to
laboratory animals: epidemiological evaluation of its potential usefulness. J. Occup. Med.
1986; 28(11): 1158-1164.

OLFERT, E.D. Allergy to laboratory animals. An occupational disease. Lab Animal 1986;
15(5): 24-31.

PIERCE, D.L. and DUKELOW, W.R. Misleading positive tuberculin reactions in a

squirrel/monkey colony. Lab. Anim. Sci. 1988; 38(6): 729-730.

RICE, D.C. Primate research: relevance to human learning and development. Dev.
Pharmacol. Ther. 1987a; 10: 314-327.

RICE, D.C. Methodological approaches to primate behavioural toxicological testing.

Neurotoxicol. Teratol. 1987b; 9: 161-169.

SCHNURRENBERGER, P.R. and HUBBERT, W.T. An outline of zoonoses. Ames, IA:

Iowa State University Press, 1981.

SCHULHOF, J. Group sets precedents for monkey bite treatment. Lab Animal 1990;
19(2): 11.

SLOVAK, A.J.M. and HILL, R.N. Does atopy have any predictive value for laboratory
animal allergy? A comparison of different concepts of atopy. Brit. J. Indust. Med. 1987;
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SOAVE, O. and BRAND, C.D. Employer responsibility for employee health in the
animal environment. Lab Animal 1991; 20(2): 41-44.

SWANSON, M.C., CAMPBELL, A.R., O'HALLAREN, M.T. et al. Role of ventilation,

air filtration, and allergen production rate in determining concentrations of rat allergens in
the air of animal quarters. An. Rev. Resp. Dis. 1990; 141: 1578-1581.

VENABLES, K.M., TEE, R.D., HAWKINGS, E.R. et al. Laboratory animal allergy in a
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WONG, J.H. and GARDELL, C. Conditioning program for non-human primates.

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 Chapter IX - Standards for Experimental Animal Surgery

IX. STANDARDS FOR EXPERIMENTAL ANIMAL SURGERY

A. INTRODUCTION

Distress resulting from inappropriate or inadequately performed surgical technique or

post-operative care constitutes "unnecessary pain". Adequate knowledge of topics such as
animal physiology, pharmacology and anatomy is essential for the success of any
research program involving the use of experimental animals, especially where surgical
techniques are required. Good surgical techniques, appropriate anesthesia, proper
instrumentation and competent pre- and post-operative care are all essential to the welfare
of the experimental animal and the success of the surgical component of the research
project, as are correctly designed surgical facilities.

All persons performing surgical techniques should have demonstrated ability in the
surgical procedures required. In this respect, it is essential that institutions provide the
opportunity for basic training and practice in required procedures before experimental
surgery is conducted. Cadaver practice and non-survival trials can help train
investigators. Adequate training and practice will help minimize anesthetic and surgical
time and contribute to faster recovery of the animal.

Medical training does not include training in the husbandry, medicine or surgery of
laboratory animals. It cannot be assumed, therefore, that prior human surgical experience
will result in good experimental animal surgery because there are significant differences
in both anesthesia and surgical technique. The guidelines of the Academy of Surgical
Research (ASR, 1989) should be consulted regarding the training necessary for the
various groups of professionals. In large experimental surgery programs, a key member
of the team should be an experienced veterinary surgeon. The primary objective is always
responsible use of the experimental animal. It is important that all personnel involved in
acute or chronic surgery treat the animals humanely and with dignity at all times. It is the
responsibility of the principal investigator to ensure that proper procedures and
precautions are observed. The following standards have been developed as a guide to this
end.

B. FACILITIES FOR SURVIVAL SURGERY

The physical environment in which surgery is performed may vary from a specially
designed, sophisticated surgery suite to a small, specifically designated area of a
laboratory. What is required will depend on the surgical procedure and whether or not the
animal is to be recovered from anesthesia. Definitions for major and minor surgery are
included in the Glossary.

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The suite in which aseptic surgery is performed should consist of the following separate
areas:

a) animal preparation area;

b) human preparation/scrub area;

c) operating room(s);

d) recovery area adequate for intensive care and post-operative support of animals;

e) support areas which would include areas for storing instruments, packs, supplies and
for washing and sterilizing instruments.

Only items used on a regular basis (e.g., anesthetic machines, suture materials, stainless
steel (s/s) kick buckets, s/s instrument tables) should be stored in the operating room
(OR). Ancillary equipment such as electrosurgery units, respirators, and
electrocardiogram (ECG) monitors should be easily sanitized, portable, and stored in the
support area if not used regularly.

It is strongly suggested that surgical facilities be within or adjacent to the animal facility.
The surgical facility should be located away from the general facility/institutional traffic.
Access to the area should be restricted to essential support staff.

The interior surfaces of the surgical facility should be impervious to moisture and easily
cleaned. Floor drains and high pressure hoses may be necessary in facilities used for large
domestic animals. The ventilation system of the OR should provide a net positive
pressure with respect to the surrounding facilities. The surgery should be supplied with
non-recirculated air. Incoming air should be as sterile as possible by means of filtration or
some other appropriate system. The operating room floor should be skid-proof. Electrical
outlets should be covered and located at least 1.5 m above the floor. The lighting in the
OR must be adequate for both surgery and clean-up. Surgical lights, either free standing
or mounted on walls or ceiling, are essential. They are equipped with sterilizable handles
so that the surgeon can adjust them. Piped-in gas services eliminate the safety hazard of
exposed pressurized tanks. Ideally services for oxygen (and nitrous oxide) and suction
should be present in the surgery, animal preparation area and in animal intensive
care/recovery. All these areas should also be equipped with a system for scavenging
anesthetic gases. Surgery tables should be durable, impervious to moisture and easily
cleaned. Stainless steel and plastic are ideal materials for this (Bennett, Brown and
Schofield, 1990).

Ideally ALL recovery surgery should be performed in a suite especially designed for this
purpose. However, it is recognized that minor (minimally invasive) surgery in small
rodents of the suborder Myomorpha (rats and mice) is often performed in laboratories. In
this case, an area in the laboratory should be set aside and used only for surgery, and

210
should be out of the main laboratory traffic. It should be uncluttered, easily cleaned, well-
lit, and should have facilities for evacuating/scavenging anesthetic gases if they are in
use. At no time should surgery be performed in an animal housing room. Major
(invasive) surgery in rodents, including stereotaxic surgery, should be performed in a
dedicated surgery room.

Intensive care/post-operative recovery should be located adjacent to the OR and close to

the persons responsible for post-operative monitoring. This area should be easily
sanitized and contain cages/pens of the appropriate size for the species being used. Cages
may vary from a sophisticated commercial unit that provides oxygen and heat, to a
standard cage in which hypothermia is prevented by increasing the environmental
temperature, e.g., by use of circulating hot water blankets, heat lamps or hot water
bottles. The type of monitoring equipment found in this room will depend on the surgery
performed. However, the means to monitor the animal's cardiovascular system,
respiratory system and core temperature should be available. An emergency kit and crash
cart should be readily available. Large domestic species (e.g., ruminants or pigs) may be
recovered in their individual stalls. The stalls should be clean, warm, dry and well-
bedded. If bedding is not in use, animals should not be recovered on the stall floor, but on
rubber mats or raised platforms. Since the stalls are likely to be at some distance from the
surgery suite, it is important that there be frequent and careful post-operative monitoring.
The recovery room should contain an area for record keeping.

C. PRE-OPERATIVE PLANNING AND ANIMAL PREPARATION

All persons involved in an experimental surgery program should be identified to ensure

that they are properly trained in the principles and practice of aseptic technique, proper
instrument use, tissue handling, closure and suturing techniques, anesthesia and
analgesia.

The primary investigator must develop a written protocol for the operative procedure in
which possible complications or special maintenance requirements arising from the
procedure are anticipated. The protocol should clearly identify the responsibilities of all
persons involved in the project; support staff, animal care staff, research technicians and
investigators. Adequate staff must be available for proper care of each animal during the
peri-operative period. For some projects, the surgical facility may need to be staffed on a
24 hour basis.

It is recommended that pre-operative care, operative technique and post-operative

care practices be developed in consultation with a veterinarian. A laboratory animal
veterinarian must be consulted to ensure that there is adequate veterinary care for the
animal, including appropriate anesthesia and analgesia.

Only healthy, disease-free animals should be used in an experimental surgery program.

Specific Pathogen Free (SPF) rodents and rabbits are available commercially. Random-
source animals must undergo a conditioning period as recommended by the laboratory

211
animal veterinarian.

A period of acclimatization, in which the animal can adjust to new environments, special
housing, tethers, slings, other forms of restraint or frequent handling, is very important.
This will greatly decrease the amount of distress or disorientation experienced by the
animal and ensure the validity of experimental results.

Surgical records should be kept for all experimental animals. The degree of detail
recorded will vary with the procedure and the species. The amount of information
recorded for a calf undergoing heart transplantation will be very different from that
recorded for a group of rats undergoing adrenalectomy, for example.

Each species has a different fasting time before surgery. Food is usually withheld for 12
hours before surgery in dogs, cats, ferrets, non-human primates (NHP) and pigs
(Flecknell, 1987). Water should be withheld only for two to three hours (if at all) before
the actual surgery so that dehydration does not result. Fasting ruminants for 24 to 48
hours prior to surgery helps to reduce the incidence of rumenal tympany (bloat)
(Flecknell, 1987). It is unnecessary to withhold food and water from rodents and rabbits
except in special circumstances such as surgery of the lower bowel.

If fasting is required, it can be done overnight in large rodents, or for up to 24 hours in

rabbits, as they retain their food longer. Mice or other small rodents with similarly high
metabolic rates should not be fasted for more than three or four hours (see also
Anesthesia).

D. SURGICAL PROCEDURES AND INTRA-OPERATIVE NURSING CARE

Methods for restraining animals for injections or the collection of body fluids are
described in Volume 2 of this Guide (CCAC, 1984). Table 1 provides a summary of the
injection sites, needle sizes and volumes to be introduced for the smaller common
laboratory species.

All species undergoing surgery should receive a similar level of care and attention.
Recovery surgery in all species of animals should be performed using aseptic
technique. Instruments should be sterile. Objects introduced into the animal, such as
telemetry implants, osmotic minipumps, vascular access ports, cannulae and any other
biomedical devices, must be sterile. Suitable preparation of the surgeon will include
wearing a scrub suit, performing a surgical scrub, wearing a cap, mask, sterile gown and
sterile surgical gloves. For minor recovery surgery in rodents, a minimum of a clean lab
coat, hand scrub, mask and sterile surgical gloves is required of the surgeon.

Surgery in field conditions should be performed in as clean an environment as possible,

with sterile instruments, sterile surgical gloves and aseptic technique.

Every effort must be made to minimize infection. The rat may exhibit increased

212
resistance to post-surgical infection compared to other rodents; however, this should not
be an excuse for less-than-adequate sterilization of implants, cannulae, etc., or for non-
sterile technique. Routine use of antibiotics is inappropriate.

Those performing "multiple run" surgeries, in which a large number of rodents are
undergoing the same procedure, should also use aseptic technique. Several sets of sterile
instruments will be required. Instruments, if used more than once, should be kept in a
germicidal solution between animals.

General publications are available that describe in detail the pre-surgical preparation of
the animal and the incision site, the preparation and sterilization of instrument packs,
drapes, fluids, etc., and the draping of the animal. For surgeries that are frequently
performed in veterinary practice (e.g., rumenotomies, thoracotomies, castrations), clinical
approaches may be used. For experimental surgery, guides to approaches for each body
system are available (Gay, 1986a, 1986b, 1989; Swindle and Adams, 1988).

TABLE 1 FOR EACH SPECIES, SITE OF INJECTION, MAXIMUM NORMALLY

ACCEPTED VOLUME AND NEEDLE SIZE*
Species Subcutaneous Intramuscular Intraperitoneal Intravenous
MOUSE Scruff, 2-3 ml, Quadriceps/posterior 2-3 ml, <21 G Lateral tail
thigh, 0.05 ml, <23 G vein, 0.2 ml,
<20 G <25 G
RAT Scruff, back, Quadriceps/posterior 5-10 ml, <21 G Lateral tail
5-10 ml, <20 G thigh, 0.3 ml, <21 G vein,
sublingual
vein, penile
vein (jugular
vein, femoral
vein--cut
down), 0.5
ml, <23 G
HAMSTER Scruff, 3-4 ml, Quadriceps/posterior 3-4 ml, <21 G Femoral or
<20 G thigh, 0.1 ml, <23 G jugular vein
(cut down),
0.3 ml,
<25 G
GUINEA Scruff, back, Quadriceps/posterior 10-15 ml, <21 G Ear vein,
PIG 5-10 ml, <20 G thigh, 0.3 ml, <21 G saphenous
vein, dorsal
penile vein,
0.5 ml, <23
G
RABBIT Scruff, flank, Quadriceps/posterior 50-100 ml, <20 Marginal ear
30-50 ml, <20 thigh, lumbar G vein, 1-5 ml,

213
 G muscles, (slowly) <21
0.5-1.0 ml, <20 G G
CAT Scruff, back, Quadriceps/posterior 50-100 ml, <20 Cephalic
50-100 ml, thigh, 1.0 ml, <20 G G vein, 2-5 ml,
<20 G (slowly), <23
G
DOG Scruff, back, Quadriceps/posterior 200-500 ml, <20 Cephalic
100-200 ml, thigh, 2-5.0 ml, <20 G vein, 10-15
<20 G G ml, (slowly),
<21 G
BIRD -- Pectoral muscles, 1-2 Midline, Brachial
(domestic ml, <21 G halfway between (wing) vein,
fowl) cloaca and 2-3 ml, <21
sternum, 10-15 G
ml, <21 G

* In intravenous administration for infusion, the amount of fluid replacement may exceed
recommended maximum volumes, particularly in dogs and cats.

Reference

TUFFERY, A.A., ed. Laboratory animals: An introduction for new experimenters. J.

Wiley & Sons Ltd. 1987

When selecting a surgical approach, it is important that the surgeon consider the anatomy
and normal body posture of the animal. This is especially important in ruminants. In this
way, the least painful approach or the one promoting a speedy recovery can be chosen.
The surgeon should also be familiar with the behaviour of the animal species being used,
so that the appropriate closure technique can be used.

During surgery, it is important that the physiological condition of the animal be

monitored and kept stable. The degree of monitoring will depend on the equipment
available. Basic monitoring of the cardiovascular system, respiratory system and core
temperature requires very little equipment. These observations should be recorded in the
animal's surgery record. It is essential that the animal be clinically examined at least
twice per day in the immediate post-operative period.

Attention should be paid to the fluid requirements of the animal. Careful attention should
be paid to hemostasis during surgery, to avoid hypovolemic shock, especially in small
animals. Prolonged surgical procedures or those in which there will be significant blood
loss require intravenous electrolyte replacement and/or blood transfusion.

The animal should be positioned on the table so as to avoid compromising cardiovascular

or respiratory function and pressure point tissue necrosis. It should be protected from

214
hypothermia and firmly, but carefully restrained in the operative position.

The use of a single animal in multiple survival surgeries is strongly discouraged. Multiple
major surgery protocols must be approved by the institution's Animal Care Committee
(ACC), and allowed only if for scientific reasons. Multiple major surgeries on a single
animal are not to be performed in order to save money. A second major surgery may
be performed if it is non-survival.

Minor procedures such as biopsies may be performed more than once. However, it is
important that animals recover completely between procedures.

The subject of anesthesia is covered elsewhere in this Guide; however, the following
points should be noted by experimental surgeons:

a) all surgical procedures are to be carried out under anesthesia;

b) those doing surgery have an obligation to be aware of the efficiency of the anesthetic
technique being used;

c) it is the responsibility of the surgeon and anesthetist to ensure that this animal is spared
discomfort during the entire peri-operative period. This includes the period during the
induction of anesthesia, for the entire surgical period and for the post-surgical recovery
period.

d) in no case is it acceptable to use muscle paralytics without appropriate

anesthetics. No ACC should approve the use of a "paralysed-awake animal" (see
also Ethics of Animal Investigation) in a surgical or other procedure which might
involve pain or distress.

E. POST-OPERATIVE RECOVERY AND SUPPORT

Recovery from anesthesia can be hazardous and requires frequent, perhaps continuous
monitoring. Depending on the anesthetic regime, recovery may take from a few minutes
to several hours. Qualified staff must be available to monitor the animal throughout the
entire recovery period. In the case of recovering neonatal rodents, care must be taken to
prevent maternal cannibalism. Under no circumstances should any animal be allowed
to recover unattended.

A number of nursing activities will be required during the immediate post-operative

period, e.g., removal of endotracheal tube if used, maintenance or removal of
intravenous lines, frequent turning of the animal to avoid bruising and vascular and
respiratory problems, and recording of physiological parameters. All these should take
place in a designated area suitable for intensive care.

When normal eating and drinking behaviour has resumed, and physiological parameters

215
have been stabilized or are within expected limits, the animal may be removed from
intensive care to more standardized husbandry. However, the animal must continue to be
monitored carefully; the wound will need attention, sutures need to be removed, catheters
flushed, etc. Depending on the model created, long-term post-operative care may
involve special diets, daily medication, physiotherapy or some other form of specialized
treatment. All animals must be monitored for signs of post-surgical infection or other
complications.

The goal of the surgery team must be to minimize any pain or distress. The degree of
post-operative pain will vary; however, in all cases, every attempt must be made to
relieve pain with appropriate use of analgesics and good nursing care. Investigators
must consult with a veterinarian to set up an analgesic regime for ALL species of
animals used. The type of analgesic, the dose and duration of treatment will depend on
the species and temperament of the animal and the type of surgery it has undergone. Most
analgesics in use are relatively short acting and require administration every few hours. It
is the responsibility of the investigator to make sure that the necessary staff are
available to administer analgesics as prescribed. The laboratory animal veterinarian
will have the necessary expertise to advise on the newer analgesics and methods of
administration.

All personnel in the project should be familiar with the animal's behaviour and
posture when normal and when in pain.

a) Responsibility for Surgical Standards

i) The responsibility for the animal in each surgical case lies with the person doing the
surgery who, in turn, should be accountable to the institutional ACC for his/her adherence
to these standards and for demonstrating an acceptable level of expertise.

ii) The responsibility for supervision of the experimental animal surgical facility should
be clearly defined.

iii) Where supportive treatment is required (analgesics, tranquillizers, antibiotics, etc.),

the surgical investigator must institute suitable treatment in consultation with a
veterinarian.

iv) If the animal, as a result of the experimental manipulation, is in distress that cannot be
relieved, authorized personnel, e.g., the laboratory animal veterinarian, should be
contacted immediately and procedures instituted for euthanasia.

F. REFERENCES

ACADEMY OF SURGICAL RESEARCH. Guidelines for training in surgical research in

animals. J. Investig. Surg. 1989; 2: 263-268.

216
BENNETT, B.T., BROWN, M.J. and SCHOFIELD, J.C. Essentials for animal research: a
primer for research personnel. Beltsville, MD: National Agricultural Library 1990.

CANADIAN COUNCIL ON ANIMAL CARE. Guide to the care and use of experimental
animals. Vol. 2. Ottawa, Ont.: CCAC, 1984.

FLECKNELL, P.A. Laboratory animal anesthesia. London: Academic Press, 1987.

GAY, W.I., ed. Methods of animal experimentation. Research surgery and care of the
research animal--Part A: patient care, vascular access, and telemetry. New York, NY:
Academic Press, 1986a; VII.

GAY, W.I., ed. Methods of animal experimentation. Research surgery and care of the
research animal--Part B: surgical approaches to the organ systems. New York, NY:
Academic Press, 1986b; VII.

GAY, W.I., ed. Methods of animal experimentation. Research surgery and care of the
research animal--Part C: surgical approaches to the organ systems. New York, NY:
Academic Press, 1989; VII.

SWINDLE, M.M. and ADAMS, R.J., eds. Experimental surgery and physiology: induced
animal models of human disease. Baltimore, MD: Williams and Wilkins, 1988.

Additional Reading

BINNINGTON, A.G. and COCKSHUTT, J.R. Decision-making in small animal soft

tissue surgery. Toronto, Ont.: B.C. Decker Inc. 1988.

BOJRAB, M.J., ed. Current techniques in small animal surgery. 3rd Ed. Philadelphia, PA:
Lea and Febiger, 1990.

CUNLIFFE-BEAMER, T.L. Surgical techniques. In: Guttman, H.N., ed. Guidelines for
the well-being of rodents in research. Bethesda, MD: SCAW (Scientists Center for
Animal Welfare), 1990.

DOUGHERTY, R.W. Experimental surgery in farm animals. 1st Ed. Ames, IA: Iowa
State University Press, 1981.

FLECKNELL, P.A. The relief of pain in laboratory animals. Lab. Anim. 1984; 18: 147-
160.

GOURLEY, I.M. and VASSEUR, P.B., eds. General small animal surgery. Philadelphia,
PA: J.B. Lippincott Company, 1985.

KNECHT, C.D., ALLEN, A.R., WILLIAMS, D.J. and JOHNSON, J.H. Fundamental

217
techniques in veterinary surgery. 3rd Ed. Philadelphia, PA: W.B. Saunders Company,
1987.

LANE, D.R., ed. Jones's animal nursing. 4th Ed. New York, NY: Pergamon Press, 1985.

MORTON, D.B. and GRIFFITHS, P.H.M. Guidelines on the recognition of pain, distress
and discomfort in experimental animals and an hypothesis for assessment. Vet. Rec.
1985; 116: 431-436.

OEHME, F.W., ed. Textbook of large animal surgery. 2nd Ed. Baltimore, MD: Williams
and Wilkins, 1988.

PIERMATTEI, D.L. An atlas of surgical approaches to the bones of the dog and cat. 2nd
Ed. Philadelphia, PA: Saunders, 1979.

PRATT, P.W., ed. Medical nursing for animal health technicians. 1st Ed. Goleta, CA:
American Veterinary Publications, Inc. 1985.

PRICE, C., ed. Practical veterinary nursing. Cheltenham, Eng.: BSAVA Publications,
1985.

ROMATOWSKI, J. Prevention and control of surgical wound infection. J. Am. Vet. Med.
Assoc., 1989; 194: 107-114.

SLATTER, D.H., ed. Textbook of small animal surgery. Philadelphia, PA: W.B. Saunders
Company, 1985.

SWINDLE, M.M. Basic surgical exercises using swine. New York, NY: Praeger
Publishers, 1983.

TRACY, D.L., ed. Small animal surgical nursing. St. Louis, MO: C.V. Mosby Company,
1983.

TUFFERY, A.A., ed. Laboratory animals: An introduction for new experimenters.

Chichester, Eng.: J. Wiley & Sons Ltd., 1987.

WAYNFORTH, H.B. Experimental and surgical technique in the rat. London: Academic
Press, 1980.

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 Chapter X - Control of Animal Research, Teaching and Testing

X. CONTROL OF ANIMAL PAIN IN RESEARCH,

TEACHING AND TESTING

A. INTRODUCTION

"Many of the advances made in our knowledge of the basic mechanisms of pain and
advances in pain therapy would have been impossible without experiments in animals,
which have yielded enormous benefits for both humans and animals. The knowledge
gained has resulted in more effective methods of pain control in both humans and
animals, has brought about a decrease in suffering, and has thus improved the quality of
peoples' lives" (Bonica, 1992).

The assessment and management of pain and suffering is a challenge that must be faced if
animals are to be treated ethically and humanely (Fosse, 1991). A landmark publication
on animal pain was Dawkins's (1980) Animal Suffering: The Science of Animal Welfare.
Recent valuable additions include Animal Pain: Ethical and Scientific Perspectives
(Kuchel, Rose and Burrell, 1990), Animal Pain (Short and Van Poznak, 1992), and a
handbook, Recognition and Alleviation of Pain and Distress in Laboratory Animals,
prepared by the Committee on Pain and Distress in Laboratory Animals of the Institute of
Laboratory Animal Resources (ILAR), which discusses stressors in the laboratory and the
animal behaviours they cause, the physiology of pain and distress, drug dosages and
euthanasia (ILAR, 1992).

The first symposium on animal pain was presented in 1982 by the Federation of
American Societies for Experimental Biology (FASEB) (Kitchell, Erickson, Carstens et
al. 1983). It was quickly followed by other publications, symposia, and guidelines related
to pain relief in animals (Zimmerman, 1983; RSPCA, 1983; Wall and Melzack, 1984;
Flecknell, 1984; Gibson and Paterson, 1985; Morton and Griffiths, 1985; AVTRW, 1986;
Frenk, Cannon, Lewis et al. 1986; AVMA, 1987; Beynen, Baumans, Bertens et al. 1987;
Rowan, 1988; Anon., 1990; Balls, 1989, 1990; Arena and Richardson, 1990; Dawkins,
1990; Goyd, 1990; LASA, 1990; Bateson, 1991; Moberg, 1992).

Sackman (1991) has prepared a review article on control of pain in cats and dogs.

B. WHAT IS ANIMAL PAIN?

In addition to ethical concerns, poor health, pain or distress in animals interject unwanted
variables into research that can greatly interfere with interpretation of the studies
(Montgomery, 1990). Pain research often requires the production of the same sensations
and behaviour in animals that ethical guidelines say must be eliminated (Amyx, 1990).
Wall (1992) suggests that instead of agonizing over an undefinable concept of pain, we
simply study the animal's efforts to stabilize its internal environment and then aid it, or at
least not intrude on those efforts without good reason.

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The question of distress in animals and how to define and measure it is still quite
perplexing (Olfert, 1992; Lewis, 1942; Brown, 1988; Molony, 1985).

Reduction or alleviation of stress or pain is considered by Flecknell (1987) as a

refinement in animal care, as part of Russell and Burch's (1959) "3Rs" of refinement,
reduction and replacement (Smyth, 1978; Rowsell and McWilliam, 1986). Poor
anesthetic techniques, for example, can adversely affect research and may produce
unnecessary pain (Flecknell, 1987). Animal suffering includes stress, distress, discomfort,
and deprivation, (Smith, 1988) as well as anxiety and fear. Freedom from discomfort and
freedom from pain, injury or disease are two of the animal's Five Freedoms, as
promulgated by the U.K.'s Farm Animal Welfare Council (FAWC) (Seamer, 1993).

In the absence of evidence to the contrary, it may be assumed that any stimuli or
experience which produces pain and discomfort in humans, also does so in animals
(LASA, 1990; RSPCA, 1983), as first promulgated by the Littlewood Committee in
1965. Amyx (1990) suggests that when Animal Care Committees (ACC) are reviewing
protocols which involve aversive stimuli, members test the stimulus on themselves.

Discomfort may not be sufficient to manifest observable pain. However, it is important to

be able to assess discomfort, because this provides the first steps towards avoiding it.

Attempts to define what constitutes stress have been made for some time, with little
agreement (Levine, 1985). However, it was recently defined by ILAR (1992) as "the
effect produced by external (i.e., physical or environmental) events or internal (i.e.,
physiologic or psychologic) factors, referred to as stressors, which induce an alteration in
an animal's biologic equilibrium." The presence or absence of stress appears to be the
only acceptable indicator of animal well-being (Duncan, 1992).

In addition to stress in the research setting, the stress of animal transportation, even for
short distances, has been demonstrated in laboratory animals (Grtner, Bttner, Dhler et
al. 1980; Clark, Mason and Moberg, 1988; Toth and January, 1990) and farm animals
(Fraser and Broom, 1990).

Sherrington (1947) originally defined a noxious stimulus as one which was actually or
potentially damaging to the skin, to which Lineberry (1981) added production of escape
behaviour in animals. The receptors specifically responsive to noxious stimuli are termed
"nociceptors" (Kitchell, Erickson, Carstens et al. 1983). However, Wall (1992) states that
there is evidence that the Central Nervous System (CNS) can extract information relevant
to pain from afferents other than specific nociceptors.

The strongest intensity of noxious stimulation that a human-being will permit is called
the "pain tolerance threshold" (Kitchell, Erickson, Carstens et al. 1983). Bateson (1991)
notes that the subjective experiences of an animal, if it has any, may be totally different
from humans, reflecting its different way of life and the different ways in which its body
works. For example, most clinical veterinary neurologists are amazed by the high pain

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thresholds of some dogs (Kitchell, Erickson, Carstens et al. 1983).

In pain research, the vast majority of animals used are rodents, specifically rats (Amyx,
1990). Silverman (1991) notes, however, that "pain detection in rodents is not easy. Slight
behavioural changes, vocalizations, abnormal use of body parts may signal pain, but we
may not be able to evaluate its magnitude." In rodents, important indicators are
recumbency and changes in the hair coat and brightness of the eyes (Montgomery, 1990).

Criteria for assessing morbid and moribund conditions in oncologic and toxicologic
research include impaired activity, change in temperament, restlessness, decreased feed or
water intake, abnormal vocalization, abnormal posture, self-mutilation and changes in
bowel or urinary activity (Montgomery, 1990).

One of the characteristics of pain or distress in animals is a change in behaviour and

reflex attributes (Amyx, 1990). Animal care personnel and research investigators must be
familiar with the normal behavioural characteristics of the experimental animal, for the
success or failure of the study can depend on the expertise of the technician observing the
animals to minimize pain and distress (Montgomery, 1990; Bateson, 1991). Moreover,
familiarity with the handler, surroundings and procedures can reduce anxiety in the
animal, as can positive re-enforcement (LASA, 1990).

C. GUIDELINES

The first code of laboratory procedures regarding animals in North America was
formulated by Walter B. Cannon in 1909, and was adopted and enforced in the
laboratories of American medical schools, and later served as the basis of American
Physiological Society's (APS) Guiding Principles in the Care and Use of Animals (Cecil
and Samuels, 1987).

In both Britain and The Netherlands, suffering is categorized as mild, moderate or

substantial in severity (Smith, 1988).

A working party of Britain's Laboratory Animal Science Association (BLASA) has

discussed the assessment and control of pain in experimental animals (LASA, 1990).
Barclay (1988) has developed a disturbance index for rodents. The Laboratory Animal
Science Association's (LASA) Working Party on Assessment and Control of Severity has
developed a Severity Index (SI). This has been applied to such areas as administration of
substances, collection of tissues and body fluids, surgical techniques and restraint. The SI
is reached by means of assigning scores based on consciousness, anesthesia, preparation
(preparatory manipulation), restraint (ranging from brief manual restraint to whole body
restraint), duration, tissue sensitivity, organ risk and mortality. As well, the consequences
in terms of pain, distress and deprivation are evaluated. Procedures are judged on a scale
of up to 34 points. Examples ranged from two points for intravenous perfusion in an
anesthetized animal, to 24 points for parabiosis (reversible suspension of obvious vital
activities or anatomical and physiological union of two organisms) (LASA, 1990).

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It is LASA's contention, based on Maslow (1970) and Curtis (1985) that interference with
the basic physiological functions or needs presents a greater risk to well-being or survival
than interference with behavioural requirements (LASA, 1990). The Canadian Council on
Animal Care (CCAC) believes, however, that at least as regards non-human primates,
"measures to safeguard psychological stability should take equal precedence to those
concerning physical health" (CCAC, 1984).

This Guide includes the Categories of Invasiveness in Animal Experiments which were
originally based on those of the Washington-based Scientists Center for Animal Welfare's
(SCAW)Categories of Biomedical Experiments Based on Increasing Ethical Concerns
for Non-Human Species (Orlans, Simmonds and Dodds, 1987). The Categories of
Invasiveness document has since been amended nine times. In the management of animal
pain, see also the CCAC statement on Ethics of Animal Investigation which is found
elsewhere in this Guide.

D. THE ROLE OF THE VETERINARIAN IN REDUCING PAIN

Veterinary training and expertise play a vital role in fulfilling an institution's

responsibilities to prevent and minimize pain and suffering in all animals used for
research, teaching and testing (Gorham, 1991; Rowsell, 1992). The Canadian Association
for Laboratory Animal Medicine (CALAM) in 1990 adopted a document--Adequate
Veterinary Care, which the CCAC considers a basis for its own policy on this topic. The
document covers the prevention and relief of animal pain.

The contribution of trained animal technicians has already been noted. The Canadian
Association for Laboratory Animal Science (CALAS) sets standards, examines and
registers laboratory animal technicians in Canada.

E. SIGNS OF PAIN AND DISTRESS*

There are numerous stereotypical responses to stress or pain stimuli in animals,

particularly in mammals. Nevertheless, species differences do exist. Recognition of
changes in behaviour and physical appearance in the species under study will allow early
identification of an animal experiencing pain or distress. Some species specific
observations are presented in this section.

Non-human Primates (NHP)

Monkeys often show remarkably little reaction to surgical procedures or to traumatic

injury. Obvious signs of pain are not readily seen. Loud and persistent vocalization, for
example, commonly signifies only alarm or anger. The animal in pain may be huddled in
a crouching posture with a "sad" facial expression and glassy eyes, or it may sit hunched
with its head forward and its arms across its body. It may avoid its companions and may
stop grooming itself. A monkey in pain may also attract increased attention from its cage
mates, which can vary from social grooming to attack. Acute abdominal pain may be
shown by facial contortions, clenching of the teeth, restlessness, and shaking

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accompanied by grunts and moans. Food and water intake is usually diminished or
absent.

Key Signs: hunched position, failure to groom, refusal of food or water, dejected
appearance.

* With acknowledgement to: Association of Veterinary Teachers and Research Workers.

Guidelines for the recognition and assessment of pain in animals. Potters Bar, Herts:
Universities Federation for Animal Welfare (UFAW), 1989; Laboratory Animal Science
Association Working Party. The assessment and control of the severity of scientific
procedures on laboratory animals. Lab. Anim. 1990; 24: 97-130; Hansen, B., Hardie, E.
and Young, M. Recognition of acute pain and distress in the dog. Humane Innovations
and Alternatives to Animal Experimentation 1990; 4: 170-173; Morton, D.B. and
Griffiths, P.H.M. Guidelines on the recognition of pain, distress and discomfort in
experimental animals and a hypothesis for assessment. Vet. Rec. 1985; 116(16): 431-436.

Dogs

Dogs in pain generally appear quieter, less alert, and withdrawn, with stiff body
movements and an unwillingness to move. In severe pain, the dog may lie still or adopt
an abnormal posture in order to minimize its discomfort. In less severe states, it may
appear restless and the immediate response to acute, but low intensity pain may be an
increased alertness. There may be inappetence, shivering, and increased respirations with
panting. Spontaneous barking is unlikely; the dog is more likely to whimper or howl,
especially if unattended, and may growl without apparent provocation. A dog may lick or
scratch at painful areas of its body. When handled, it may be abnormally apprehensive or
aggressive. The animal exhibits anxious glances; it seeks cold surfaces. Its tail is often
between its legs.

Penile protrusion and frequent urination may also be noted.

Key Signs: inappetence, bites at pain regions, abnormally apprehensive.

Cats

Cats in pain are generally quiet, with an apprehensive facial expression; the forehead may
appear creased. There may be crying or yowling and the cat may growl and hiss if
approached or made to move. There is inappetence and a tendency to hide or to separate
from other cats. The posture becomes stiff and abnormal, varying with the site of the
pain. A cat with head pain may keep its head tilted. If the pain is generalized in the thorax
and abdomen, the cat may be crouched or hunched. With thoracic pain alone, the head,
neck, and body may be extended. In abdominal or back pain, the cat may lie in lateral
recumbency with its back arched. If the animal is standing or walking, the back is arched
and the gait stilted. Incessant licking is sometimes also associated with localized pain.

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Pain in one limb usually results in limping or holding up of the affected limb.

A cat in severe pain may show demented behaviour and make desperate attempts to
escape. If a painful area is touched or palpated, there may be an instant and violent
reaction. There may be panting, with an increased pulse rate and pupillary dilatation. A
cat in chronic pain may have an ungroomed appearance and show a marked change from
its normal behaviour. The animal exhibits tucked in limbs, hunched head and neck, and
utters a distinctive cry or hissing and spitting sound. Its ears are flattened. It shows fear of
being handled and may cringe.

Key Signs: stiff posture, demented behaviour, lack of grooming, hunched head and
neck, inappetence.

Mice

After procedures which cause pain, mice may increase their sleeping times. Reduced food
and water intake, with resultant weight loss, dehydration and wasting of the muscles on
the back may be observed. Piloerection (erection of hair) and a hunched appearance
indicate pain or distress. The animal fails to groom, but scratches more frequently. Sick
mice are often isolated from the remainder of the group. Aggressive vocalization is
observed in the early stages, decreasing where pain or stress reduces the ability to move
and respond.

The eyes appear sunken, and ocular and nasal discharge may be noted as the animal's
condition worsens. The respiration rate increases and breathing may be forced or
laboured. Defecation/urination are immediate reactions to stress in the mouse, and
increase or decrease as stress continues. The movement of vibrissae (muscle hairs)
becomes less evident as pain or stress continues. Affected mice become more timid and
apprehensive; however, as pain or stress increases, they may become aggressive, with a
tendency to bite. The animal may attempt to bite the source of pain or affected area, and
may self-mutilate the affected part.

Writhing movements are noted when the pain is abdominal. There is gradual assumption
of a hunched, 'sleeping posture' away from any light source. Where limbs or feet are
affected, sudden running movements are exhibited as an escape mechanism; there is
increasing difficulty in maintaining posture. The mouse may show unsteady gait,
difficulty in moving in straight line, and circling movements where balance is affected. A
rolling gait is often noted with developing ascites.

As its condition worsens, the animal becomes quiet and unresponsive, separates from the
group and eventually becomes unaware of its surroundings. Hypothermia is observed
with increasing deterioration in condition; the animal feels 'cold' to the touch.

Key Signs: withdrawal, biting response, piloerection, hunched back, sunken eyes
and abdomen, dehydration, weight loss.

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Rats

Rats are generally docile and less aggressive than mice towards members of their own
species and humans. Acute pain or distress is usually accompanied by constant
vocalization and struggling. Rats will often lick or guard a painful area. Increased
scratching can indicate chronic pain. A rat in pain will often sit crouched with its head
turned into its abdomen. Sleeping periods will be disturbed and increase if pain or
distress are present. An elevated respiratory rate associated with sneezing occurs where
the respiratory system is affected. Increasing piloerection (staring coat) is noted, along
with an increasingly untidy appearance as the animal fails to groom itself. There may be
some hair loss. The animal ceases to eat and drink normally. There is poor skin tone, and
evidence of muscle wasting along the back--indicative of dehydration and weight loss.

During repeated painful or distressing procedures, animals may become more aggressive
and resist handling, which will increase with increasing pain or distress. The eyelids
rapidly assume a half-closed or almost-closed position. The eyes may appear sunken, and
ocular discharge is common, often progressing to red-coloured hematoporphyrin exudate
which may encircle the eye. Nasal discharge, if present, may be red-coloured as well
(Harkness and Ridgway, 1980).

Constipation or diarrhea may occur depending on the organ system(s) affected. Urination
decreases with reduced water intake; however, frequency may increase where urinary
infection or hormonal disturbance is present. Animals in pain initially show increased
awareness/aggressive responses and a tendency to bite, but eventually become depressed
and unresponsive. Exploratory behaviour lessens. Aversive behaviour is shown towards
other animals. There is possible self-mutilation of affected parts in later stages.
Abdominal contraction and stilted movements may occur if abdominal pain is present.
There may be increasing pain associated with locomotion. Lameness in one of the limbs
or simply careful gait may be noted. A "waddling" gait occurs where abdominal
enlargement take place as a result of intestinal obstruction or ascites. Circling often
occurs where balance is disturbed.

Initially, the rat exhibits increased angry or aggressive vocalization, especially on

handling. There is a gradual reduction in vocal response as the pain or stress continues,
and movement ceases unless a sudden painful stimulus is experienced. Hypothermia
indicates significant deterioration in the animal's condition. A pale appearance indicates
anemia or blood loss.

Key Signs: vocalization, struggling, licking/guarding, weight loss, piloerection,

hunched position, hypothermia.

Guinea Pigs

Guinea pigs are alert, but timid and apprehensive animals which will try to avoid capture
and restraint. Rarely is there any aggression towards humans. Any sign of acceptance
indicates the animal is unwell. Loud vocalization will accompany even minor and

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transient pain. Guinea pigs often appear sleepy when in pain. Initially, there is an
increased level of response to painful or stressful stimuli. However, this gradually
subsides and the animal becomes unresponsive. It gradually appears more apprehensive.
The eyes may be sunken and dull. The respiratory rate increases as a painful or stressful
stimulus increases or continues; where the respiratory system is affected, respirations
become increasingly forced and laboured. Often loss of weight occurs as well as hair loss,
scaly skin, and dehydration. Where the gastrointestinal tract is affected there may be
evidence of diarrhea. There is a tendency to 'barbering' under dietary stress with failure to
eat or drink. Group aggression may occur and damage to the skin of the back may result
from fighting. There is excessive salivation where abnormal teeth cause eating
difficulties, a tendency to an arched back where abdominal pain is present, and failure of
the "righting" reflex in seriously ill animals. There may be pain associated with
locomotion, lameness, and careful gait due to sore feet in older animals.

Key Signs: withdrawal, vocalization, failure to resist restraint, staring coat,

unresponsive.

Mongolian Gerbils

Gerbils are highly active, nervous animals and usually attempt to avoid restraint. Signs of
pain and distress are difficult to assess, as gerbils apparently object to any interference.
There is an increased level of response under painful or stressful stimuli. Ocular
discharge is common. Under stressful conditions, the eyelids may be half closed, with dry
matting of the eyelids. The increased respiratory rate associated with lung involvement is
difficult to assess by eye. Loss of coat condition occurs. Loss of hair from the tail may be
seen in overcrowded animals. Facial lesions and sores may result from excessive
burrowing in the corners of the cage.

Dehydration is rarely seen, since the gerbil's normal metabolism enables full utilization of
the water content of the diet. Only small quantities of urine are voided under normal
conditions. Feces are normally firm, dry pellets. Constipation is rare. Diarrhea, if it
occurs, may quickly lead to death from fluid loss. Gerbils are normally extremely active
and nervous. Under severe stress, there may be temporary collapse and apparent shock
syndrome; however, the animals recover, given time. Changes in exploratory behaviour
and increased aggressive response may occur. A hunching up and arching of the back
may be observed, especially with abdominal involvement. Abnormal gait is associated
with locomotion or abdominal involvement.

Key Signs: hunched appearance, weight loss, shock syndrome.

Syrian (golden) Hamsters

Under normal conditions, hamsters will sleep for long periods during the day, and little
activity will be seen. They often appear aggressive towards their cage mates and emit
loud screeching noises, disproportionate to the degree of interference, when handled. This
response increases under painful or stressful stimuli. Ocular discharge is commonly

226
associated with stress. An increased respiratory rate is associated with lung involvement.
Loss of coat condition is seen where the diet is deficient in Vitamin E and short chain
fatty acids. Loss of body condition occurs with decreased food and water intake.
Constipation is unusual in the hamster. Diarrhea, when it occurs, is profuse and liquid,
staining the perineal region. Increasing depression takes place when the animal is left
undisturbed. Daytime sleep periods may be extended and increasing lassitude may be
seen except when the animal is being handled. Exploratory behaviour is reduced. A
hunched appearance is noted, as is an unwillingness to move, especially where abdominal
organs are involved. Lateral recumbency can indicate that the animal is moribund.
Normal gait is affected when pain is associated with locomotion. Stilted movements are
sometimes associated with abdominal involvement, e.g., ascites following cirrhosis of the
liver.

Key Signs: weight loss, hunched appearance, increased aggression or depression,

extended sleep periods.

Rabbits

The rabbit presents significant difficulties in recognition of pain and distress, as it often
quietly accepts apparently painful or distressing procedures; this may relate to its feral
behaviour where concealment is important to survival. Even healthy rabbits may not
move frequently or indulge in exploratory behaviour. Pain is usually characterized by a
reduction in food and water intake (and thus weight loss and dehydration) and limited
movement. Although rabbits frequently become ill and distressed without showing much
apparent loss of condition, careful examination will reveal a loss of muscle mass on the
lower back. Ocular discharge is a common response to stress in the rabbit, with
protrusion of the nictitating membrane.

Under continued pain or stress, rabbits assume a 'sleepy' appearance. The animal exhibits
increased depression, progressive unawareness and lack of response. The animal will
often face the back of cage, away from light. An increased respiratory rate is associated
with either apprehension or lung involvement. There is fecal staining of the coat. Night
time pellet production may be interrupted. Constipation and diarrhea are common
responses to pain or stress. Excessive self-grooming may precipitate hair balls in the
stomach. Where foot soreness is involved, weight may be thrown forward or backward to
reduce discomfort. Body stretching and lying flat are common indications of abdominal
discomfort. Pain may be associated with locomotion, especially with sore feet.

Key Signs: reduced eating and drinking, faces towards back of cage, limited
movement, apparent photosensitivity.

Horses

Periods of restlessness are noted in horses experiencing pain or distress. Food is held in
the mouth uneaten. The horse exhibits an anxious appearance with dilated pupils and
glassy eyes; increased respiration and pulse rate with flared nostrils; profuse sweating and

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a rigid stance. In prolonged pain, behaviour may change from restlessness to depression
with the head lowered. In pain associated with skeletal damage, limbs may be held in
unusual positions and there is a reluctance to move, with the head and neck "fixed."
There may be a pain-induced tachycardia.

In abdominal pain, a horse may look at, bite or kick its abdomen; it may get up and lie
down frequently; walk in circles; or roll. When near collapse, the horse may stand very
quietly, rigid and unmoving, but with signs of deteriorating circulatory status such as
mucosal cyanosis and prolonged capillary filling time. Horses in pain generally show a
reluctance to be handled.

Key Signs: anxious appearance, restlessness, biting at site of pain, depression, fixed
position.

Cattle

Cattle in pain often appear dull and depressed, with the head held low and showing little
interest in their surroundings. There is inappetence, weight loss and, in milking cows, a
sudden drop in milk yield. Severe pain often results in rapid shallow respirations. On
being handled, they may react violently or adopt a rigid posture designed to immobilize
the painful region. Grunting and grinding of the teeth may be heard. Acute pain may be
associated with bellowing. Generally, signs of abdominal pain are similar to those seen in
the horse, but are less marked. Rigid posture may lead to a lack of grooming due to an
unwillingness to turn the neck. In acute abdominal conditions, such as intestinal
strangulation, the animal adopts a characteristic stance with one hind foot placed directly
in front of the other. Localized pain may be indicated by persistent licking of an area of
skin, or kicking at the offending area.

Key Signs: dull, depressed, inappetence, grunting, grinding of the teeth, rigid
posture.

Sheep and Goats

In general, signs of pain in these species are similar to those in cattle. Changes in posture
and movement are apparent, and a change in facial expression may be indicative of pain
or distress. There is a general reluctance to move. Goats are more likely than cattle to
vocalize in response to pain. Grinding of the teeth and grunting are also heard. Sheep, in
particular, tolerate severe injury without overt signs of pain or distress. Following
procedures such as castration and tail docking, lambs may show signs of discomfort such
as standing up and lying down repeatedly, tail wagging, occasional bleating, neck
extension, dorsal lip curling, kicking, rolling and hyperventilation.

Key Signs: rigid posture and reluctance to move.

Pigs

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Pigs in pain may show changes in gait and posture. They normally squeal and attempt to
escape when handled; however, these reactions may be accentuated when the animal is in
pain. Adult pigs may become aggressive. Squealing is also characteristic when painful
areas are palpated. Handling of chronic lesions may not elicit signs of pain. Pigs will
often be unwilling to move and may hide in bedding if possible.

Key Signs: vocalization and the lack of normal social behaviour may be helpful
indicators of a pig in pain.

Birds

Birds in pain may show escape reactions with vocalization and excessive movement.
Head movements increase in extent and frequency. There may be an increase in heart and
respiratory rates. Prolonged pain will result in inappetence and inactivity with a drooping,
miserable appearance. The eyes may be partially closed, the wings held flat against the
body, and the neck retracted. When handled, the escape reaction may be replaced by a
state of tonic immobility. Birds with limb pain will avoid use of the affected limb and will
"guard" it from extension.

Key Signs: escape reactions, atonic immobility, inappetence, avoidance of use of

pain site.

Reptiles

Acute pain in reptiles may be characterized by flinching and muscle contractions. There
may be aversive movements away from the unpleasant stimulus, and attempts to bite.
More chronic and persistent pain may be associated with anorexia, lethargy and weight
loss, although it is difficult to associate any of these signs of lack of well-being
specifically with pain.

Key Signs: flinching and muscle contractions, weight loss, anorexia.

Fishes

It is difficult to determine the nature of the response to pain in fish. Although they exhibit
a pronounced response to injuries or to contact with irritants, their response to chronic
stimuli may be small or absent. Fish with severe wounds which would cause immobility
in a mammal, will often appear to behave completely normally, even resuming feeding.
Fish will react to noxious stimuli, such as that administered by a hypodermic needle, by
strong muscular movements. When exposed to a noxious environment, such as a strong
acid, they show abnormal swimming behaviour with attempts to jump from the water,
their colouring becomes darker and their opercular movements become more rapid. Such
effects are indicative of some degree of distress; however, it is not possible to describe
these unequivocally as signs of pain.

F. ANALGESIC AGENTS

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The appropriate use of analgesics during or after a painful procedure is an integral part of
a protocol plan. The following general information, as well as details of administration
and dosages by species, are given in several references (Sawyer, 1985; Sackman, 1991;
Flecknell, 1984) in addition to the anesthetic textbooks listed in the additional reading.

The opioids (morphine-like drugs) are the most widely used analgesic agents. Opioids act
by binding to specific receptors. The main classes of receptors are m.

1. Opioid Agonists

Opioids produce potent hypnotic and analgesic effects including significant depression of
the cardiovascular and respiratory systems and an alteration in the thermoregulatory
mechanism. The euphoria and addiction associated with opioids in human is not a
problem in animals when the drugs are used properly. Some opioids induce vomiting in
dogs and NHP and rapid intravenous injection may occasionally result in an excitatory
phase in most species. In farm animals, as well as in the cat and mouse, the effects of
opioids are less predictable, and undesired excitement may occur. Avoidance of the
excitement phase in species with an enhanced sensitivity to opioids can often be achieved
by the use of very low dosages (Green, 1982).

Opioids used in the veterinary medicine include morphine, meperidine, fentanyl,

oxymorphone, etorphine (M99) and carfentanil. They are pure or relatively pure m
agonists and are all good analgesics.

a) Morphine is most frequently used clinically for the control of post-operative pain in
dogs and NHP, providing up to four hours of pain relief. In dogs, its use is complicated by
undesirable gastrointestinal effects. Intravenous bolus administration in dogs may cause
histamine release which may contribute to morphine's hypotensive action (Hall and
Clarke, 1991). As a pre-medicant, morphine's stimulatory effect on the vagus nerve may
induce bradycardia, unless atropine is given in advance. Profound respiratory depression
occurs rapidly and is dose-related. Intracranial and intra-ocular pressure are increased
(Sackman, 1991).

b) Meperidine has effects similar to morphine and is the drug of choice for
premedication in the dog, as very little gastrointestinal stimulation is induced. However,
severe hypotension may occur after intravenous use. As analgesia lasts only one to two
hours, it is not recommended for alleviating post-surgical pain. This drug has also proven
useful as a post-operative sedative for NHP and horses.

c) Fentanyl is a very potent short-acting opioid. It is combined with droperidol to make a

neuroleptanalgesic that provides profound analgesia. New synthetic compounds of
fentanyl include alfentanil, which has an ultrashort half-life, and sufentanyl with a half-
life shorter than fentanyl, but with fewer peripheral side effects (Flecknell, 1984).

d) Oxymorphone is more potent than morphine and produces more sedation in the dog

230
than morphine or meperidine. Post-surgical pain relief lasts two to six hours.
Cardiovascular stability is much greater than with the other opioids. It is frequently
combined with diazepam or acepromazine for anesthesia and analgesia in old or sick
animals. An anticholinergic should be given to prevent severe bradycardia.

e) Etorphine (M99) is an extremely potent morphine derivative with a high tendency to

produce initial excitement followed by depression. Because of its potency, it has been
widely used in dart guns for the capture and immobilization of zoo animals and wild
game (Fowler, 1986; Green, 1982). The drug has also been used successfully in certain
cold blooded animals (Fowler, 1986; Green, 1982). It is extremely dangerous to humans;
therefore, diprenorphine (M5050) must be available for immediate reversal should
accidental human exposure occur (Fowler, 1986).

f) Carfentanil is currently preferred to etorphine by many zoo veterinarians because of

its higher potency, which allows administration by swabbing or spraying of the buccal or
nasal mucosa. It can be reversed by cyprenorphine (M285) or diprenorphine (M5050)
(Lumb and Jones, 1984) and naltrexone. Carfentanil can be fatal in humans if
accidentally injected (partial immobilizing dose).

2. Opioid Agonist/Antagonists

The search for analgesic agents with fewer side effects than pure m agonists led to the
development of partial m agonists and kappa agonists such as butorphanol and
buprenorphine. This group of drugs may also be used to reverse the depressant effects of
an opioid while preserving the analgesic qualities.

a) Butorphanol is a synthetic analgesic with five times the potency of morphine. A

degree of sedation occurs and the respiratory depression has a ceiling effect that does not
increase with higher doses. Cardiovascular effects are minimal and it is a poor opioid
antagonist (Dyson, 1990). Analgesia lasts two to five hours following
subcutaneous injection, and may be accompanied by some dysphoria.

b) Buprenorphine is a long-acting analgesic that antagonizes the depressant effects of

the opioid agonists, while still maintaining long-term post-operative analgesia of 8-12
hours in many species (Flecknell, 1984).

c) Pentazocine lactate is a poor analgesic with a very short duration (approximate half-
life in the dog of 22 minutes). It has minimal cardiovascular effects and is a mild
respiratory depressant.

d) Nalbuphine is slightly less potent than morphine, with a wide safety margin and
minimal cardiovascular and respiratory depression. Analgesia lasts three to eight hours. It
has also been used as an opioid antagonist to reverse sedation and respiratory depression
of opioids while maintaining analgesia (O'Hair, Dodd, Phillips et al. 1988).

3. Opioid Antagonists

231
Naloxone hydrochloride, an effective antagonist, is available to reverse the effects of
opioids (this includes the analgesia). It has no agonist properties and does not produce
respiratory or cardiovascular depression. It has an antagonistic effect for one to four
hours, and can be used to reverse the effects of any of the opioid agonist/antagonist
group. Nalorphine or diprenorphine must be available when using etorphine, in case of
accidental human administration (Lumb and Jones, 1984). Naltrexone is a long-acting
derivative of naloxone. At present, its use in veterinary medicine is limited; however,
should a pure long-acting antagonist be required, it could prove useful (Hall and Clarke,
1991).

4. Non-steroidal Anti-inflammatory Drugs (NSAIDS)

These agents produce analgesia by reducing inflammation and thus peripheral

sensitization. They have little, if any, central analgesic action. Side effects are
interference with platelet and renal function and gastric ulceration. Cats metabolize these
agents slowly and must be dosed infrequently to prevent toxicity. Those commonly used
in dogs are the carboxylic acid group (aspirin, naproxen, meclofenamic acid, flunixin)
and the enolic acids (phenylbutazone, dipyrone, and piroxicam). In cats aspirin,
phenylbutazone and dipyrone are frequently used at low dosages (Sawyer, 1985).

a) Aspirin relieves pain associated with peripheral inflammation, but is ineffective for
visceral pain. In cats, it should be given only every 48 to 72 hours.

b) Naproxen is used when aspirin does not relieve the pain, and the once daily
administration is convenient. As is the case with aspirin, gastric ulcers are listed as a side
effect.

c) Meclofenamic acid is popular for treating musculoskeletal pain that is refractory to

aspirin. It has 1.5 times the potency of phenylbutazone.

d) Flunixin is reported to have greater analgesic properties than phenylbutazone,

meperidine or codeine, and is used for osteoarthritic pain. Its use in large animals is well
established. Its use in small animals is expanding. In dogs, it has the potential to produce
serious gastrointestinal effects (bleeding) if a maximum of three doses is
exceeded (Hall and Clarke, 1991).

e) Phenylbutazone relieves musculoskeletal pain, but has been associated with blood
dyscrasias, gastrointestinal disturbances, nephropathies and hepatitis.

f) Dipyrone is an analgesic, antipyretic, anti-inflammatory that also may cause blood

dyscrasias with prolonged use.

g) Piroxicam is popular in human medicine for its once daily dosage and very effective
relief of osteoarthritic pain. Toxicity is similar to that of other non-steroidal anti-
inflammatory drugs (Sackman, 1991).

232
5. Analgesia provided by Local Anesthetics

As an alternative to systemically administered agents, analgesia can be provided by use

of local anesthetics. Bupivacaine, a long-acting local anesthetic, is preferred for post-
operative analgesia (Flecknell, 1992). It can either be injected around specific nerve
trunks which supply the surgical site, or infiltrated into the muscular and subcutaneous
tissue layers during closure of a surgical incision. Use of local anesthesia to infiltrate the
surgical wound is a simple technique that can provide 4-12 hours analgesia.

Selective blocking of the intercostal nerves two to three intercostal spaces on either side
of a thoracotomy incision has recently been recommended for relief of pain following
thoracotomy in dogs. Bupivicaine hydrochloride is used before incision closure, and
provides four to five hours of analgesia. The respiratory pattern of dogs recovering from
thoracotomies is not significantly changed. This provides a distinct

advantage over opioid analgesics which may cause significant respiratory depression.
This technique, while providing relief of pain associated with the surgical incision, does
not elevate visceral (intrathoracic) pain (Sackman, 1991).

6. Neuroleptanalgesics

Neuroleptanalgesia is a state of sedation and analgesia produced by the combined use of a

tranquillizer (neuroleptic) and an opioid. Minor surgery can be performed; however, the
patient remains rousable and responds to certain stimuli. Moderate respiratory depression
occurs, and muscle relaxation tends to be poor, but can be counteracted by combination
of the neuroleptanalgesic with a benzodiazepine (Flecknell, 1987). The most commonly
used preparation is Innovar-Vet, (droperidol 20 mg/ml and fentanyl 0.4 mg/ml), which
should not be confused with Innovar, prepared for human use (droperidol 2.5 mg/ml and
fentanyl 0.0005 mg/ml).

Innovar-Vet has been used extensively in the dog and is reported useful in many other
species. It has a wide margin of safety, is well tolerated by animals in poor physical
condition, and is partially reversible with naloxone. Significant bradycardia may be
avoided by prior use of atropine. Its use is contra-indicated in the cat, cow, horse and
sheep due to CNS stimulation. Animals remain responsive to auditory stimuli, and
aggressiveness during recovery and other disposition changes lasting several days have
been reported in the dog (Lumb and Jones, 1984).

A variety of other opioids and tranquillizers can be combined to produce

neuroleptanalgesia; among these, mixtures of morphine/promazine and
etorphine/acepromazine have proven useful in a variety of animals (Flecknell, 1987).
Meperidine/acepromazine (Lumb and Jones, 1984) and oxymorphone/acepromazine
(Short, 1987) have also been used for the dog and cat.

G. AREAS FOR FUTURE STUDY

233
Farm animal welfare, transgenic animals, amphibians, reptiles and invertebrates all
constitute areas of increasing concern.

The welfare of domestic animals is of great importance, and it is claimed by Spira (1986)
that, because of their numbers, 95% of all animal suffering takes place in intensive
management ("factory farming") practices; thus, every 1% reduction in their suffering
will accomplish more than all other protection campaigns for other species of animals put
together. Animal behaviour and applied animal ethology are increasingly becoming areas
of great importance (Maxie, 1987; Fraser, 1988; Fraser and Broom, 1990; McKeown and
Luescher, 1988; Duncan, 1992).

Another area that will become of increasing concern is the production of transgenic
animals (Jaenisch, 1988; Baker, 1988; Ewing, 1990; Cross, 1990; McLaren, 1990; Page,
1990) and the possible pain and distress that may be caused them. Their uses in research
have recently been discussed (Saffer, 1992; Merlino, 1991) as well as management of
their colonies (Geistfeld, 1991). The CCAC, foreseeing the need for guidelines on animal
biotechnology, has recently established a committee comprising knowledgeable
scientists, and representatives of industry and animal welfare, to develop such guidelines
to include embryo manipulation, fetal research and transgenic animals. Agriculture
Canada is also considering a proposed framework for regulating the production and use
of transgenic animals (Sethi, 1992).

Only recently has the analgesia, anesthesia and euthanasia of amphibians, reptiles and
fish been addressed (UFAW, 1989; Johnson, 1992; Iwama, 1992; Davis, 1992).
Evaluation of pain and stress has been discussed in reptiles (Lance, 1992), cold-blooded
vertebrates (Arena and Richardson, 1990; Fiorito, 1986), and birds (Gentle, 1992). It has
been contended that fish can experience pain and fear to a degree which can be compared
with human reactions (Anon., 1988).

In addition to studies involving vertebrates, the use of invertebrates in research is

governed in Canada by CCAC's Categories of Invasiveness found elsewhere in this
Guide. It states that cephalopods and some other higher invertebrates have nervous
systems as well developed as some vertebrates and therefore may warrant inclusion in the
Categories under B, C, D and E. In Great Britain, only one species of cephalopod, the
common octopus (Octopus vulgaris), is only now being brought under the Animals
(Scientific Procedures) Act (Anon., 1993). Handling, anesthesia and surgery of
cephalopods are outlined in a recent Universities Federation for Animal Welfare
publication (Boyle, 1991).

Other areas of concern to the scientific community and ACCs include, for example, the
effects of blood loss (McGuill, 1989) and the use of Freund's Complete Adjuvant (FCA)
(Broderson, 1989). An expert committee of CCAC has been established especially to
examine this latter issue and investigate replacements for FCA. The CCAC Guidelines on
Acceptable Immunological Procedures appear elsewhere in this Guide, and are revised as
new knowledge becomes available.

234
In the development of research in the future, the scientist should consider the importance
of refinement and should address those studies which are known to cause the most pain
and suffering (Rowsell, 1992).

We have a considerable distance to go before animals need no longer be used. However,

as Medawar (1972) notes: "We must grapple with the paradox that nothing but research
on an animal will provide us with the knowledge that will make it possible for us, one
day, to dispense with the use of them altogether." And to paraphrase Wall (1984): "So
long as one animal remains in pain and we cannot help, our knowledge of pain remains
inadequate."

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 Chapter XI - Anesthesia

XI. ANESTHESIA

This chapter provides guidance and information on anesthesia and relief of pain in
experimental animals. It is not meant to be comprehensive, and non-veterinary users
should consult with a veterinary anesthesiologist or laboratory animal veterinarian when
such drugs are to be administered. Information on common dosages and means of
administration of analgesic, tranquillizing and anesthetic agents are given in the
Appendices. The agents described in this chapter are all prescription and/or
controlled drugs. Non-veterinary users may obtain prescription drugs from a
licensed veterinarian, and should contact the Bureau of Dangerous Drugs, Health
and Welfare Canada regarding the use of controlled drugs in research.

Methods for assessing the depth of anesthesia vary with the species and the drug, and are
discussed in Green (1982). Specific details are available in the textbooks and review
articles listed in the references.

A. MANAGEMENT OF ANESTHESIA

1. General

Sedatives, analgesics, and general anesthetic agents must be utilized for the control of
pain and distress unless contrary to the achievement of the objectives of the study. In the
latter case, approval of the institutional Animal Care Committee (ACC) is
mandatory.

Anesthetic agents frequently affect the cardiovascular, respiratory and thermoregulatory

mechanisms, in addition to the central nervous system (CNS). Every effort should
therefore be made to maintain the circulation, respiratory function and the body
temperature of the anesthetized subject within normal physiological limits (Parker and
Adams, 1978). Endotracheal intubation ensures that the airway remains patent and free
from obstruction.

Hypothermia may occur during exposure to anesthetic gases and during intra-abdominal
surgery, particularly in small animals. This may result in death or a greatly prolonged
recovery from the anesthetic. The degree of hypothermia may be reduced by placing the
animal on a circulating warm water blanket or other device that assists in conserving
body heat (Muir and Hubbell, 1989; Lumb and Jones, 1984; Flecknell, 1987).

2. Handling the Patient

The animal should always be handled gently and calmly in order to minimize
struggling and fright. Prolonged excitement will disturb the circulatory and metabolic
state of the patient and induce a degree of shock. Furthermore, attempts to anesthetize a
struggling animal present physical problems in addition to enhancing the likelihood of an
abnormal response to the anesthetic agents. These points are particularly important when

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restraining and anesthetizing wild animals (Fowler, 1986).

3. Fasting

Cats, dogs, non-human primates (NHP), ferrets and pigs should receive no food during
the 8-12 hours prior to induction of anesthesia in order to minimize the risk of vomiting
during induction or recovery from anesthesia (Flecknell, 1987). Very small or immature
mammals should be subjected to a much shorter fast, usually from two to four hours, due
to their higher metabolic rate. Withholding food from ruminants for 12-24 hours may
help reduce the incidence of ruminal tympany (bloat); however, reduction of the volume
of digesta in the rumen requires much longer periods of starvation (36-72 hours). Water
should be withheld for 12 hours before surgery to prevent gorging and increase in the
volume of rumen contents. Pre-anesthetic fasting of small rodents or rabbits is
unnecessary since they do not vomit during induction (Flecknell, 1987). Guinea pigs
should be fasted 6-12 hours before anesthesia to allow time to clear their mouths of the
food bolus commonly carried at the base of the tongue. Small birds often are not fasted at
all, in order to maintain energy during the stress of the procedure (Muir and Hubbell,
1989; NRC [U.S.], 1977). Fasting pregnant animals of all species, particularly ruminants,
can produce severe metabolic disturbances. Other than ruminants, every animal should be
provided with drinking water until approximately one hour before induction of anesthesia
(Flecknell, 1987).

4. Anticholinergics

Anticholinergics block parasympathetic stimulation to the cardiopulmonary system and

reduce salivary secretion. They are used in combination with sedatives and analgesics as
pre-medication to general anesthesia. Anticholinergics are no longer routinely
administered to each animal undergoing anesthesia. They are administered selectively,
after a pre-anesthetic clinical examination of the animal, and according to the determined
needs of the individual patient, the anticipated response to the anesthetic medication, and
the tendency to develop bradycardia or excessive salivation (Short, 1987).

a) Atropine is the most commonly used anticholinergic agent; however, routine

administration is controversial due to the high incidence of associated cardiac
dysrhythmias (premature ventricular contractions and sinus tachycardia) (Lumb and
Jones, 1984; Flecknell, 1987). It is most commonly recommended for use in NHP, pigs,
guinea pigs and chinchillas in order to decrease airway secretions, but should not be
given if a marked tachycardia is already present (Green, 1982).

b) Glycopyrrolate is a quaternary ammonium anticholinergic. Although its mechanism

of action is similar to that of atropine, its effects last longer. Glycopyrrolate seems to be
less likely than atropine to produce sinus tachycardia (Paddleford, 1988). It does not
penetrate the CNS because of its difficulty in crossing the blood-brain barrier. It is also
less likely than atropine to cross the placental barrier, indicating that it is a selective
peripheral anticholinergic agent (Short, 1987).

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B. TRANQUILLIZERS AND SEDATIVES

Tranquillizers produce a calming effect without sedation (Green, 1982). They have no
analgesic properties, and even at the high doses that cause ataxia (failure of muscular co-
ordination) and depression, animals are easily aroused. Tranquillizers are useful over a
wide range of species, often in combination with other drugs, to lessen the dose of a
general anesthetic and produce a smoother induction and recovery. Sedatives are used to
produce drowsiness and reduce fear and apprehension (Flecknell, 1987).

The psychological state of the animal prior to administration of tranquillizers may

markedly affect the degree of sedation achieved. Animals that are vicious, intractable and
in a state of excitement may not become manageable except with very high
(incapacitating) doses.

a) Phenothiazines (promazine, acepromazine) produce sedation and reduce the dose of

drugs needed for general anesthesia, but also cause moderate hypotension and
hypothermia (Lumb and Jones, 1984; Flecknell, 1987).

b) Benzodiazepines (diazepam, midazolam) produce variable sedation depending on

the species (Lumb and Jones, 1984; Flecknell, 1987; Green, 1982). They are good muscle
relaxants and have no marked undesirable side effects. Diazepam cannot be mixed with
other water soluble agents, while midazolam can (Flecknell, 1987).

c) Butyrophenones (azaperone, droperidol) have similar effects as phenothiazines, but

are more potent and cause less hypotension (Lumb and Jones, 1984; Flecknell, 1987;
Green, 1982). Droperidol is used in combination with an opioid to produce
neuroleptanalgesia (Flecknell, 1987).

d) alpha-2-adrenergic agonists (xylazine, detomidine, medetomidine)

i) Xylazine (Rompun) is a sedative and analgesic that acts as a CNS depressant and
induces muscle relaxation by inhibiting the transmission of impulses in the CNS. Its
major use in laboratory animal anesthesia is in combination with ketamine to produce
surgical anesthesia. This combination has been used in dogs, cats, NHP, large farm
animals and wild animals (Olson and McCabe, 1986; Lumb and Jones, 1984). It causes
respiratory depression and a bradycardia which may progress to heart block (Flecknell,
1987). It also increases the susceptibility of the myocardium to circulating
catecholamines during halothane anesthesia (Short, 1987). Vomiting may occur in dogs
and cats, and gas accumulation due to gastrointestinal atony (lack of normal tone or
strength) may be a problem in both large dogs and ruminants (Lumb and Jones, 1984).
Xylazine produces profound physiological changes and its safe use requires knowledge
of these effects which are often species specific. Yohimbine and 4-aminopyridine reverse
most of the effects of xylazine without relapse in many species (Jernigan, Wilson, Booth
et al. 1988), with the exception of NHP (Lynch and Line, 1985).

ii) Detomidine is marketed for use in horses, and has the same cardiovascular effects

244
(bradycardia and hypotension) as xylazine, but is more potent and has a longer-acting
effect.

iii) Medetomidine is being evaluated for use in dogs and cats, and has cardiovascular
effects similalr to xylazine. A medetomidine/ketamine combination in cats has the
advantage over xylazine/ketamine in that a lower dose of ketamine is needed, the
duration of action is longer and the analgesia better (Verstegen, Fargetton, Donnay et al.
1990).

C. GENERAL ANESTHETICS

1. Dissociative Anesthetics

Dissociative anesthetics produce a state of chemical restraint and anesthesia characterized

by muscle rigidity and dissociation of the mind from the external environment. The eyes
remain open, necessitating use of protective ointment. Various reflexes, including the
blinking reflex and laryngeal reflex, remain intact, and adequate respiration is normally
maintained. An increase in heart rate, blood pressure and intracranial pressure frequently
occurs. Thus, their use is contra-indicated in head injuries or intra-ocular surgery. While
the use of dissociative anesthetic agents is most common with NHP and cats, they have
also been used in most other mammalian species as well as birds and reptiles (Jones,
1977). Combination with a tranquillizer is recommended in most species to enhance
analgesia and reduce muscle tone (Flecknell, 1987; Green, 1982).

a) Ketamine hydrochloride is the most commonly used member of this group. Depth of
anesthesia is dose related. Side effects include excessive salivation which may be
controlled with atropine (Flecknell, 1987), a tendency toward convulsions, and a recovery
characterized by excitement, disorientation, and hallucinations which may be controlled
by tranquillizers and barbiturates (Lumb and Jones, 1984). In all cases, a smooth recovery
will be facilitated if the patient is left undisturbed in a quiet, darkened environment.

b) Tiletamine is similar to ketamine, but is longer lasting and more potent; therefore, a
smaller dose volume is needed. It is most commonly sold in combination with the
tranquillizer zolazepam (Telazol), which improves muscle relaxation, CNS depression,
and emergence from anesthesia. It also prevents tiletamine seizures. Cats may take 12-36
hours to be clinically "normal" following tiletamine anesthesia. Tiletamine/zolazepam has
proven successful in rats and gerbils, but not in mice or hamsters (Hrapkiewicz, Stein and
Smiler, 1989). Tiletamine causes nephrotoxicity in rabbits (Brammer, Doerning, Chrisp
et al. 1991; Doerning, Brammer, Chrisp et al. 1992).

2. Barbiturates

Barbiturates differ from tranquillizers and opioids in that increasing the dose
progressively increases the depth of depression until a state of general anesthesia is
reached. They are poor analgesics. Their primary use is in the induction and/or
maintenance of general anesthesia. Barbiturates are potent respiratory depressants and

245
their effects on the cardiovascular system are variable. At intermediate dosages,
excitement is sometimes induced (Green, 1982).

The barbiturates are grouped according to duration of action into long acting (e.g.,
phenobarbital), short- or intermediate-acting (e.g., pentobarbital) and ultrashort-acting
(e.g., thiopental, thiamylal, methohexital) (McLaughlin, 1988). The short- and ultrashort-
acting drugs are commonly used for anesthesia. Anesthetic duration varies widely with
species; however, in general, short/intermediate barbiturates produce approximately 2-3
hours of anesthesia and ultrashort barbiturates range from 10 to 20 minutes (McLaughlin,
1988).

Variation in dose response and duration of effect of barbiturates is extreme within and
between species (Olson, 1986a; Green, 1982; McLaughlin, 1988). The following are
examples of the variation found with pentobarbital (intermediate) anesthesia:

i) cats frequently having a considerably prolonged sleeping time (McLaughlin, 1988);

ii) mice on hardwood bedding take almost twice as long to recover as mice on softwood
bedding, and male mice sleep longer than female mice (McLaughlin, 1988);

iii) the anesthesia produced in adult horses and cattle is of relatively short duration;
however, the recovery period is long and difficult (Lumb and Jones, 1984).

Whenever possible, barbiturates should be administered intravenously, slowly, to effect.

Administration by other routes is far less satisfactory, as dosage is more difficult to judge
and the anesthetic effects are less predictable. Any of the barbiturates can cause skin
sloughing if perivascular injection accidently occurs (McLaughlin, 1988).

Although barbiturates are commonly used, they are often poor choices for general
anesthesia due to poor analgesia, profound cardiovascular effects, high mortality and
numerous external factors that can affect dose response and sleeping time. Adequate
anesthesia can be obtained by combining a barbiturate with a tranquillizer, sedative or an
opioid (Olson, 1986a; Lumb and Jones, 1984; McLaughlin, 1988).

3. Chloralose

Chloralose may be used for non-survival experiments requiring prolonged anesthesia

and minimal surgical interference (Flecknell, 1987; Holzgrefe, Everitt and Wright,
1987). There is disagreement about whether chloralose is a true anesthetic agent or a
hypnotic with little analgesic action. It is used primarily for physiological studies to
preserve the vagal and central baroreceptor reflexes or in acute cardiovascular studies to
preserve myocardial function. While chloralose is generally considered to have no
application in survival studies or in clinical veterinary medicine (Lumb and Jones, 1984),
one recent study used chloralose repeatedly over a long time period in puppies without
any signs of toxicity (Grad, Witten, Quan et al. 1988).

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4. Urethane (Urethan, Ethyl Carbamate)

Urethane produces long periods of anesthesia, has a wide safety margin and little effect
on normal blood pressure and respiration. It produces sufficient analgesia to allow
surgical manipulations (Flecknell, 1987). However, the drug should be handled with
extreme care as it is considered to be cytotoxic, carcinogenic and
immunosuppressive. It also causes profound changes in gastrointestinal function and is
stimulatory to the hypothalamus and pituitary (Olson, 1985). Animals should not be
allowed to recover following urethane anesthesia.

5. Saffan

Saffan is a combination of two steroids, alphaxalone and alphadolone dissolved in a

surfactant (vehicle), Cremaphor EL, to solubilise it. It is administered intravenously or
intramuscularly, although the latter route gives more unpredictable results. Muscle
relaxation is good, and recovery rapid. It is rapidly metabolized and is an excellent agent
for long-term maintenance (Flecknell, 1987). It has been used for the cat, pig, large farm
animals, small NHP, rodents, birds and exotics (Lumb and Jones, 1984; Flecknell, 1987;
Green, 1982). It is not recommended in the dog due to the associated massive histamine
release caused by the Cremaphor EL vehicle that often occurs (Flecknell, 1987). Saffan
must not be used with barbiturates (Flecknell, 1987).

6. Tribomoethanol (Avertin)

The use of Avertin is controversial because of the wide variation in results between
laboratories. Although no longer available in Canada, it may be introduced in a different
formulation. Purchased as a powder, it must be dissolved in amylene hydrate and then
diluted with distilled water at 40C immediately prior to use. Great care must be taken to
use only fresh solutions as it decomposes very rapidly in light or temperatures above
40C, producing byproducts that are severe tissue irritants. In rodents, it is given
intraperitoneally (Green, 1982), resulting in good muscle relaxation and moderate
respiratory and cardiovascular depression (Flecknell, 1987; Green, 1982); however, post-
operative fatalities are often high due to peritoneal adhesions. Even if a freshly prepared
solution is used, mortality is often high after administration of a second anesthetic at a
later date (Green, 1982; Norris and Turner, 1983).

7. Non-specific Injectable Anesthetic Antagonists

Several agents have the ability to reverse many of the effects of non-opioid injectable
anesthetics through non-specific antagonistic properties.

a) Yohimbine blocks central alpha-2-adrenoreceptors, and partially antagonizes

barbiturates, xylazine, ketamine, benzodiazepines and phenothiazines (Fowler, 1986;
Lumb and Jones, 1984).

b) 4-aminopyridine (4-AP) partially antagonizes xylazine, ketamine and barbiturates.

247
Yohimbine and 4-AP are often combined for a more effective reversal (Lumb and Jones,
1984).

c) Doxapram is a respiratory stimulant and not a reversal agent per se; however, it has
been used to partially antagonize the respiratory depression produced by barbiturate
anesthesia in dogs (Hatch, Jernigan, Wilson et al. 1986).

8. Inhalant Anesthetics

Inhalant anesthetics have the advantage of requiring minimal detoxification by the body,
as they are exhaled through the lungs, and the level of anesthesia can be easily and
rapidly controlled. However, their use requires specialized equipment for administration,
and constant monitoring of the patient (Stimpfel and Gershey, 1991). Some are explosive
or inflammable, or tissue irritants. Chronic exposure to some agents is hazardous to the
health of the operating room personnel (Lumb and Jones, 1984).

The speed of induction and recovery depend on the solubility of the anesthetic in blood.
Highly soluble anesthetics (methoxyflurane) are slow to reach an equilibrium in the
blood; therefore, induction and recovery are prolonged. Insoluble anesthetics (halothane)
reach an equilibrium rapidly, making manipulation of anesthetic depth easier, but also
more hazardous due to the potential for rapid overdose (Flecknell, 1987).

The use of inhalation anesthesia requires the following equipment:

i) a vaporizer for the volatile anesthetics;

ii) a source of carrier gas (usually oxygen or air);

iii) a breathing system from which the anesthetic mixture is inhaled;

iv) a mask or endotracheal tube for connecting the breathing system to the patient
(Sedgwick and Jahn, 1980; Gilroy, 1981). Exceptions are discussed with the individual
agents. Numerous simple systems have been devised and reported in the laboratory
animal literature for use in small laboratory animals (Dudley, Soma, Barnes et al. 1975;
Skartvedt and Lyon, 1972; Rich, Grimm, Wong et al. 1990; Olson, 1986b; Levy, Zwies
and Duffy, 1980; Mulder and Hauser, 1984).

Unnecessary exposure of personnel to gases from volatile anesthetics must be avoided by

use of appropriate scavenger systems (Muir and Hubbell, 1989). Several reports have
suggested a health risk associated with prolonged and repeated exposure to low
concentrations of halothane (hepatocellular toxicity), methoxyflurane (renal toxicity),
nitrous oxide (neurologic disease and pernicious anemia) and to the chronic ingestion of
chloroform (renal and hepatic tumours in rodents) (Rettig, 1987; Stimpfel and Gershey,
1991). Expired gases should be vented to the exterior or adsorbed onto activated charcoal
(Mitchell, 1976).

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a) Ether-based Volatile Agents

i) Diethyl ether is a highly volatile agent of relatively low potency and wide range of
safety. Ether produces good muscle relaxation and analgesia; however, it is very irritating
to mucous membranes. The vapours are highly explosive, necessitating extreme caution
in its use and storage. Due to the risk of explosion, the use of ether is discouraged as
excellent alternatives are now available (Flecknell, 1987; Stimpfel and Gershey, 1991).

ii) Methoxyflurane (Metofane) is a highly soluble, potent ether-based anesthetic.

Because of its low volatility, it may be used safely for induction with anesthetic
chambers, and nose cone maintenance. Methoxyflurane produces some respiratory and
cardiovascular depression, but less than halothane at comparable depths of
anesthesia. Myocardial sensitization occurs, but is not as severe as with halothane.
Muscle relaxation and analgesia are good, and it is neither irritating nor explosive in
anesthetic concentrations. In animals, methoxyflurane anesthesia for less than one hour is
not usually associated with hepatorenal toxicity, especially if periods of hypoxia and/or
hypercapnia are avoided (Stimpfel and Gershey, 1991).

iii) Enflurane provides rapid induction and emergence from anesthesia. It provides
moderate levels of analgesia and muscle relaxation, the latter decreasing as anesthetic
concentrations increase. It produces profound depression of respiratory functions and
myocardial performance (Short, 1987). It is largely eliminated via the lungs. Unlike
halothane, very little of the drug is metabolized by the liver. This may offer some
experimental advantages; otherwise, there is little to choose between enflurane and
halothane in terms of efficacy (Flecknell, 1987). Enflurane is expensive and requires a
special vaporizer.

iv) Isoflurane is less potent than halothane or methoxyflurane. It is relatively insoluble

which leads to fast inductions and recoveries. It may be used in halothane vaporizers that
have been recalibrated. It produces a slightly more severe respiratory depression than
does halothane, but slightly less depression of the cardiovascular system (Flecknell,
1987). There is very little myocardial sensitization to catecholamines; in fact, isoflurane
has the greatest margin of safety with the cardiovascular system of all the inhalant
anesthetics. Isoflurane produces better muscle relaxation than halothane, but has poorer
analgesic properties. It undergoes even less biotransformation than enflurane and is
almost completely eliminated in exhaled air (Flecknell, 1987). Isoflurane has a pungent
odour which may cause breath holding during induction. It has no known toxicities, but it
is expensive (Raper, Barker, Burwen et al. 1987).

b) Halogenated Hydrocarbons

i) Halothane, a halogenated hydrocarbon, is highly potent and volatile. It should be used

only with a finely calibrated precision vaporizer. It produces dose-dependent depression
of the cardiopulmonary system and hypotension (Flecknell, 1987). There is direct
myocardial depression and sensitization to circulating catecholamines. The analgesia

249
offered by halothane is reasonable, as is muscle relaxation. The vapours are neither
explosive nor irritating, but can be hepatotoxic to man (Lumb and Jones, 1984).

c) Other Agents

i) Nitrous oxide has very low anesthetic potency. Induction of a state of general
anesthesia or even unconsciousness is not possible in most animal species (Flecknell,
1987; Mahmoudi, Cole and Shapiro, 1989). As it exerts minimal effects on the
cardiopulmonary system, it can be used to reduce the required concentration of other
agents and so reduce the degree of depression at a particular depth of anesthesia
(Flecknell, 1987). It has some analgesic properties in animals; however, the potency is
less than half that experienced in humans (Short, 1987). Following cessation of nitrous
oxide administration, 100% oxygen must be administered to the
animal to prevent hypoxia caused by the rapid diffusion of the gas from the body
(Flecknell, 1987; Short, 1987). Because it presents numerous occupational hazards,
nitrous oxide should be scavenged. If a carrier gas is required 100% oxygen is effective
and non-toxic as well as being vital to life (Stimpfel and Gershey, 1991).

D. MUSCLE RELAXANTS

1. Glyceryl Guiacolate

Glyceryl guiacolate (guaifenesin) is a centrally acting muscle relaxant, with its action on
the internuncial neurons of the spinal cord. As the drug has little effect on the diaphragm,
it produces muscle relaxation without respiratory paralysis. A state of sedation and
hypnosis is produced; however, the degree of analgesia is in dispute. Guaifenesin is most
often used as part of induction technique in large farm animals. It is useful in
combination with thiobarbiturate for short surgical procedures and for intubation prior to
the administration of an inhalant anesthetic (Lumb and Jones, 1984). Guaifenesin has
been added to ketamine and xylazine to produce effective anesthesia in ponies, dogs and
pigs with minimal cardiovascular and respiratory depression. This same combination has
also been used in a continuous infusion for long term anesthesia in cats (Brown,
McCarthy and Bennett, 1991).

2. Neuromuscular Blocking Agents

Succinylcholine (a depolarizing agent), curare, pancuronium, gallamine, atacurium and

vecuronium (non-depolarizing agents) are neuromuscular blocking agents which act
peripherally at the neuromuscular junctions. Anticholinesterases such as neostigmine,
pyridostigmine and edrophonium are antagonistic to the non-depolarizing agents, but
ineffective against the depolarizing agents (Lumb and Jones, 1984). Neuromuscular
blocking agents are used as adjuncts to general anesthetics where profound muscle
relaxation is desired.

These agents produce motor paralysis only. There is no sedation or analgesia. Their
use on conscious animals is prohibited (see also Ethics of Animal Investigation).

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The use of neuromuscular blocking agents abolishes some of the signs used to judge the
depth of anesthesia. Autonomic functions remain intact with the newer agents (atacurium,
vercuronium); therefore, increases in heart rate and arterial blood pressure may indicate
the perception of pain. Animals must be artificially ventilated as the respiratory muscles
are paralyzed. Should neuromuscular blocking agents be a component of an anesthetic
protocol, it is extremely important that proper equipment and personnel with experience
in the use of these agents be available.

E. LOCAL AND REGIONAL ANESTHETICS

Local anesthetics such as lidocaine, procaine, bupivacaine and tetracaine may be used to
block the nerve supply to a limited area for the performance of minor or rapid procedures.
Local anesthesia is also frequently used as an adjunct to various sedative and hypnotic
agents in more prolonged and invasive procedures, such as caesarian section. Local
anesthetic agents may be used for the regional infiltration of a surgical site, field
blocking, nerve blocks, and for epidural and spinal anesthesia (Green, 1982; Elmore,
1981; Kero, Thomasson and Soppi, 1981; Gray and McDonell, 1986). Veterinary
assistance should be sought in the initial use of the last three procedures (Lumb and
Jones, 1984; Gray and McDonell, 1986). A combination of lignocaine/prilocaine has also
been used topically for pain-free venipuncture in some laboratory animals (Flecknell,
Liles and Williamson, 1990).

F. ANIMAL HYPNOSIS (Tonic Immobility)

A state of hypnosis or tonic immobility can be readily induced in a variety of animals

including rabbits, birds, small rodents and reptiles (Prestrude and Crawford, 1970;
Danneman, White, Marshall et al. 1988). It is characterized by a lack of spontaneous
movement or overt response to external stimuli for up to several minutes, and is usually
exhibited under stressful or fearful conditions. There is evidence that animals remain
aware of external events and hypnosis can be interrupted by mild tactile or auditory
stimuli. It is usually induced by placing the animal on its back and gently extending the
neck and hind legs to place traction on the spine. Recent work indicates that some degree
of analgesia is produced with hypnosis; however, individual animal susceptibility to
hypnosis varies greatly and in consequence hypnosis cannot be recommended as a
suitable alternative to appropriate analgesics when painful procedures are to be
performed (Danneman, White, Marshall et al. 1988).

G. SPECIES CONSIDERATIONS

a) Canine

General anesthesia: sedation, followed by intravenous induction with an ultrashort-acting

barbiturate, intubation and maintenance with an inhalant anesthetic. Alternatively,
intermediate or long-acting barbiturates may be used but are poor analgesics and can
result in profound respiratory and cardiovascular depression (Flecknell, 1987; Green,

251
1982). Minor surgical procedures can be carried out using neuroleptoanalgesics, xylazine
combinations and diazepam combinations (Green, 1982).

b) Feline

General anesthesia: sedation, induction using an injectable agent, intubation and

maintenance with an inhalant anesthetic (Green, 1982). The larynx should be sprayed
with a local anesthetic such as 2% lidocaine (without epinephrine) prior to intubation
(Flecknell, 1987). Mask induction with an inhalant anesthetic is also well tolerated if the
cat is sedated previously and handled expertly. Ketamine and ketamine combinations
have proven very useful for restraint and minor surgical procedures (Flecknell, 1987;
Ingwersen, Allen, Dyson et al. 1988). Saffan or xylazine also produce sedation and
anesthesia for minor surgical procedures (Flecknell, 1987; Green, 1982).

c) Ferrets

Administration of intravenous drugs can be difficult in the awake ferret; therefore,

alternate routes are usually used. Intramuscular ketamine and ketamine combinations are
useful (Muir and Hubbell, 1989; Moreland and Glaser, 1985), as are fentanyl/ droperidol
and intravenous Saffan (Flecknell, 1987; Green, 1982). For induction with inhalation
anesthetics, a special induction chamber is usually used, with maintenance by mask or
intubation (Poole, 1987; Moody, Bowman and Lang, 1985).

d) Rabbits

Neuroleptanalgesics and ketamine combinations with xylazine, acepromazine or

azaperone have been used successfully (Muir and Hubbell, 1989; Olson, 1986a;
Flecknell, 1987; Lipman, Marini and Erdman, 1990). Ketamine alone does not produce
adequate anesthesia or analgesia (Lumb and Jones, 1984; Flecknell, 1987). The degree of
analgesia produced by Saffan is generally low. At the higher dose rates needed to produce
medium or deep surgical anesthesia, there may be sudden apnea followed by cardiac
arrest (Flecknell, 1987). A technique of continuous intravenous infusion of ketamine and
xylazine has been reported to maintain a light anesthetic plane for up to 4 hours, although
hypoxemia and hypotension are marked (Wyatt, Scott and Richardson, 1989). Inhalant
anesthetics and mask induction are readily tolerated (Peeters, Gil, Teske et al. 1988).
Endotracheal intubation in the rabbit is relatively difficult for anatomical reasons.
Barbiturates alone are not recommended in rabbits, as the dose required to produce
surgical anesthesia is very close to the lethal dose. Respiratory arrest frequently occurs
before the onset of surgical anesthesia. They may be used, if combined with a sedative or
tranquillizer (Olson, 1986a; Peeters, Gil, Teske et al. 1988). If atropine is used it must be
at high dose levels to counteract the presence of serum atropinase (Muir and Hubbell,
1989).

e) Small laboratory rodents (rats, mice, guinea pigs, gerbils, hamsters and wild
rodents)

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Withholding food and water is unnecessary prior to anesthesia, since vomiting normally
does not occur (Flecknell, 1987). Anesthetic agents used include barbiturates, ketamine,
ketamine combinations (Muir and Hubbell, 1989; Flecknell, 1987; Wixson, 1987a,
1987b), neuroleptoanalgesics (Muir and Hubbell, 1989; Green, 1982; Parkes, 1987;
Olson, 1986a), tiletamine/zolazepam (Muir and Hubbell, 1989) and Saffan (Green, 1982).
Ketamine alone produces severe respiratory depression at doses high enough for surgical
anesthesia in small rodents (Flecknell, 1987). Intramuscular ketamine/xylazine causes
muscle necrosis in Syrian hamsters and is not recommended in that species (Gaertner,
Boschert and Schoeb, 1987). The same problem has been noted with fentanyl/droperidol
in guinea pigs (Holmes, 1984). Ketamine combinations and pentobarbital are poor
anesthetics in the gerbil, but fentanyl/metomidate (Flecknell, John, Mitchell et al. 1983)
and tiletamine/zolazepam have proven effective (Hrapkiewicz, Stein and Smiler, 1989).
Barbiturates are still in common use, but are very poor analgesics, and often cause high
mortality, especially when given intraperitoneally or when full-strength commercial
solutions are used intravenously (dilution is recommended). When combined with a
sedative, tranquillizer or an opioid, adequate anesthesia results (Olson, 1986a).

Induction of anesthesia with an inhalational agent is best accomplished with an induction

chamber. Anesthesia may be maintained with a face mask. Endotracheal intubation is
difficult in small rodents and requires purpose-made laryngoscopes (Flecknell, 1987).

The safe administration of general anesthesia to the guinea pig is notoriously difficult,
since they often maintain their pedal reflex and make squirming movements even when
deeply anesthetized (Holmes, 1984). Their response to many injectable anesthetics is very
variable. Post-anesthetic complications such as respiratory infections, digestive
disturbances, and generalized depression, are seen (Flecknell, 1987). Spinal anesthesia
offers a useful alternative (Green, 1982).

Very brief procedures (e.g., orbital blood sampling) may be performed on rodents by
using a 50:50 mixture of carbon dioxide and oxygen, if the animal is removed from the
gas chamber as soon as the pedal reflex has disappeared (Green, 1982; Fenwick and
Blackshaw, 1989).

Hypothermia may be used to anesthetize neonatal mice and rats (1-2 days old). The pup
is placed in an ice water slush for 20-30 minutes (Green, 1982).

f) Non-human Primates

Ketamine and its combinations are most often used for restraint, particularly where rapid
recovery is desired. Neuroleptanalgesics have also been used, and Saffan is useful for
small species such as marmosets. The NHP can be intubated and inhalation anesthesia
administered using techniques similar to those used for the human (Flecknell, 1987;
Sainsbury, Eaton and Cooper, 1989).

g) Horses

253
Both induction and recovery from anesthesia may be associated with excitement. Due to
their size and strength, special facilities are required for induction and recovery in horses.
Veterinary consultation should be sought. Xylazine and acepromazine are most
commonly used as pre-anesthetics, followed by an induction agent (thiamylal sodium,
guaifenesin, etc.) and inhalation anesthesia (Muir and Hubbell, 1989; Green, 1982).

h) Ruminants

Many surgical procedures can be performed under local or regional anesthesia (Muir and
Hubbell, 1989; Green, 1982; Gray and McDonell, 1986). The greatest problems with
sedation and general anesthesia are regurgitation, hypoventilation and bloat. The use of
atropine in ruminants is controversial, as it induces bloat and increases the viscosity of
the saliva, while not decreasing the quantity. Xylazine is given at one-tenth the dose of
horses, and usually results in recumbency. It is probably best to administer xylazine by
slow intravenous injection in sheep and goats as results produced by intramuscular
administration are unpredictable. It should be noted that some goats appear particularly
sensitive to xylazine (Hall and Clarke, 1991). Bloat is often a problem following xylazine
administration, and abortion can be induced in the last trimester (Muir and Hubbell,
1989). Xylazine/ketamine with or without guaifenesin can be combined for shorter
surgical procedures (Coulson, Januszkiewicz, Dodd et al. 1989). Other recommended
injectables for sheep are Saffan and ketamine/diazepam (Flecknell, 1987).

Although thiopental is useful for induction, pentobarbital is not recommended, especially

in goats, due to respiratory depression. Animals less than three months old metabolize
barbiturates very poorly (Muir and Hubbell, 1989).

Mask induction with an inhalant anesthetic such as halothane or isoflurane is particularly

useful with the smaller species. Sheep should always be intubated to prevent aspiration if
regurgitation occurs. Intubation is accomplished with the use of a laryngoscope in small
ruminants and by direct palpation of the larynx in large ones. A lidocaine spray should be
used on sheep vocal cords prior to intubation to prevent laryngospasm (Flecknell, 1987).

i) Swine

Pigs should be fasted 12 hours before surgery to prevent vomiting; however, water can be
offered until the pre-anesthetic is given (Muir and Hubbell, 1989). As respiratory
depression is a frequent sequela to general anesthetics in pigs, reversible drugs such as
xylazine or opioids are recommended (Green, 1982; Muir and Hubbell, 1989). Epidural
anesthesia is also commonly used (Muir and Hubbell, 1989). Ketamine in combination
with xylazine, diazepam, acepromazine or fentanyl/droperidol have produced good
results as general anesthetics (Muir and Hubbell, 1989; Green, 1982; Swindle, 1985), as
have other injectable anesthetic such as Saffan (Flecknell, 1987) and
tiletamine/zolazepam (Muir and Hubbell, 1989; Bauck, 1984; Cantor, Brunson and
Reibold, 1981). Barbiturates are generally used only in combination with a sedative
(Muir and Hubbell, 1989). Azaperone produces sedation, but has no analgesic effect
(Flecknell, 1987).

254
Inhalation agents are well tolerated and mask induction can be carried out on small pigs
with ease (Becker, 1986). Intubation of the trachea is difficult for anatomical reasons
(Lumb and Jones, 1984; Flecknell,1987; Green, 1982), and a lidocaine spray on the vocal
cords is used to prevent laryngospasm (Green, 1982).

Malignant hyperthermia has been observed in response to inhalation anesthetics

(especially halothane), depolarizing muscle relaxants and stress in swine. A
predisposition to hypothermia is probably inherited (Basrur, Bouvet and McDonell,
1988), and is commonest in the Landrace and Poland China breeds. Dantrolene is
effective therapy for malignant hyperthermia (Muir and Hubbell, 1989).

j) Avian

Hypothermia is a frequent problem in general anesthesia, especially for small birds.

Small birds are also prone to handling shock, and small friable vessels make intravenous
injection difficult (Green, 1982). Ketamine is an effective pre-anesthetic, and
ketamine/xylazine (Muir and Hubbell, 1989) or ketamine/diazepam (Fowler, 1986) are
two of the safest injectable anesthetics. Tiletamine/zolazepam is an alternative to
ketamine/xylazine (Muir and Hubbell, 1989; Green, 1982). Diazepam combined with
chloropent (chloral hydrate, sodium pentobarbital, magnesium sulfate) provides surgical
anesthesia for 60-90 minutes in the domestic fowl (Christensen, Fosse, Halverson et al.
1987). Saffan has been used in a variety of avian species (Lumb and Jones, 1984).
However, it should only be administered intravenously, and even then used with great
caution due to associated cardiac arrhythmias (Green, 1982; Short, 1987).

Inhalant anesthesia with mask induction can be used fairly safely and effectively;
however, because of the efficiency of the avian respiratory system, changes in anesthetic
depth tend to occur very rapidly, especially in small birds (Muir and Hubbell, 1989;
Lumb and Jones, 1984; Green, 1982). Resuscitation is complicated due to accumulation
in the air sacs (Fowler, 1986; Ludders, Mitchell and Schaefer, 1988). Inhalants cannot be
used for thoracic procedures because the gas leaks through the opened air sacs
(Christensen, Fosse, Halverson et al. 1987), and positive pressure ventilation is necessary
for abdominal procedures due to an incomplete diaphragm. Restraint must allow free
movement of the sternum for respiration. Isoflurane is the safest inhalation anesthetic,
followed by halothane (Muir and Hubbell, 1989).

k) Cold Blooded Animals

Agents commonly used include tiletamine/zolazepam, ketamine, Saffan, tricaine

methanesulfonate (MS-222) and inhalant anesthetics. Dosage varies widely between
species. Absorption and excretion of injectable anesthetics are directly related to
environmental temperature.

Fish should be fasted 24-48 hours to prevent vomiting (Green, 1982). They are
commonly anesthetized by immersion or use of a recirculation system that passes an

255
anesthetic solution over the gills. Tricaine methanesulfonate (MS-222) (Brown, 1987),
and benzocaine (Green, 1982) are recommended, although numerous other anesthetics
including carbon dioxide, ether, chloral hydrate, halothane and Saffan have also been
used (Muir and Hubbell, 1989; Lumb and Jones, 1984; Green, 1982). Benzocaine is as
effective as MS-222, equally safe for personnel and much less expensive (Green, 1982).
Exposure of benzocaine to direct sunlight causes breakdown and releases highly toxic
chlorine (Poole, 1987).

Reptiles and amphibians can be effectively anesthetized with local anesthetics, immersion
in a solution containing an anesthetic agent, injectable or inhalation anesthetics (Muir and
Hubbell, 1989). Hypothermia should only be used for restraint in non-painful procedures,
as it is not known whether or not analgesia is induced. Secondary tissue damage also
results from the practice. Hypothermia is not a suitable anesthetic for major surgery
(Muir and Hubbell, 1989). Amphibians can be anesthetized by immersion in MS-222,
which provides excellent muscle relaxation and analgesia (Muir and Hubbell, 1989;
Green, 1982). Preferred injectable anesthetics for reptiles include ketamine and
tiletamine/zolazepam, although Saffan and etorphine have also been used successfully
(Muir and Hubbell, 1989; Fowler, 1986).

Inhalation anesthesia is induced by soaking a cotton ball with a volatile anesthetic and
placing it with the animal in a box or bag, or using an induction chamber or face mask
(Muir and Hubbell, 1989). Halothane, isoflurane and methoxyflurane are preferred to
ether (Muir and Hubbell, 1989). Reptiles are relatively easy to intubate, as the larynx is
readily visualized. Their slow respiratory rates and ability to breath-hold constitute
complicating factors (Muir and Hubbell, 1989). Inhalants are not recommended for
turtles (Green, 1982).

Johnson (1992) warns that in administering anesthetics to amphibians and reptiles, one
must consider the structure of the reptilian respiratory system. Respiratory movements
are different in snakes, which have one lung, crocodiles which have diaphragms, and
lizards which have pleuroperitoneal cavities. He suggests that, because their respiratory
movements may be weak, if a volatile anesthetic is used, one may have to assist
respiration because they have a poor way of expelling air. Johnson also notes that, if
anesthesia is to be done for a long period of time, amphibians must be kept moist; as they
are all poikilotherms, keeping them at their preferred optimum temperature zone will
have an effect on the absorption and excretion of the anesthetic.

l) Invertebrates

Volk (1986) discusses methods of evaluating anesthetic depth in various invertebrates and
includes a complete list of anesthetics.

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BECKER, M. Anesthesia in Gottingen miniature swine used for experimental surgery.

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BRAMMER, D.W., DOERNING, B.J., CHRISP, C.E. and RUSH, H.E. Anesthetic and
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BROWN, L.A. Recirculation anaesthesia for laboratory fish. Lab. Anim. 1987; 21: 210-
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BROWN, M.J., MCCARTHY, T.J. and BENNETT, B.T. Long-term anesthesia using a
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CANTOR, G.H., BRUNSON, D.B. and REIBOLD, T.W. A comparison of four short-
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CHRISTENSEN, J., FOSSE, R.T., HALVERSEN, O.J. and MORILD. I. Comparison of

various anesthetic regimens in the domestic fowl. Am. J. Vet. Res. 1987; 48: 1649-1657.

COULSON, N.M., JANUSZKIEWICZ, A.J., DODD, K.T. and RIPPLE, G.R. The
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DANNEMAN, P.J., WHITE, W.J., MARSHALL, W.K. and LANG, C.M. An evaluation
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DOERNING, B.J., BRAMMER, D.W., CHRISP, C.E. and RUSH, H.E. Nephrotoxicity of
tiletamine in New Zealand white rabbits. Lab. Anim. Sci. 1992; 42: 267-269.

DUDLEY, W.R., SOMA, L.R., BARNES, C., SMITH, T.C. and MARSHALL, B.E. An
apparatus for anesthetizing small laboratory animals. Lab. Anim. Sci. 1975; 25: 481-482.

ELMORE, R.G. Food-animal regional anesthesia. Porcine blocks: lumbosacral

(epidural). VM SAC 1981; 76: 387-388.

FENWICK, D.C. and BLACKSHAW, J.K. Carbon dioxide as a short-term restraint

anesthetic in rats with subclinical respiratory disease. Lab. Anim. 1989; 23: 220-228.

FLECKNELL, P.A., JOHN, M., MITCHELL, M. and SHUREY, C. Injectable anesthetic

techniques in 2 species of gerbils (Meriones libycus and Meriones unguiculatus). Lab.

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Anim. 1983; 17: 118-122.

FLECKNELL, P.A. Laboratory animal anesthesia. Toronto, Ont.: Academic Press, 1987.

FLECKNELL, P.A., LILES, J. and WILLIAMSON, H. The use of lignocaine-prilocaine

local anesthetic cream for pain-free venipuncture in laboratory animals. Lab. Anim. 1990;
24: 142-146.

FOWLER, M.E. Zoo & wild animal medicine. Toronto, Ont.: W.B. Saunders Co., 1986.

GAERTNER, D.J., BOSCHERT, K.R. and SCHOEB, T.R. Muscle necrosis in Syrian
hamsters resulting from intramuscular injections of ketamine and xylazine. Lab. Anim.
Sci. 1987; 37: 80-83.

GILROY, B.A. Endotracheal intubation of rabbits and rodents. J. Am. Vet. Med. Assoc.
1981; 179: 1295.

GRAD, R., WITTEN, M.L., QUAN, S.F., MCKELVIE, D.H. and LEMEN, R.J.
Intravenous chloralose is a safe anesthetic for longitudinal use in beagle puppies. Lab.
Anim. Sci. 1988; 38: 422-425.

GRAY, P.R. and MCDONELL, W.N. Anesthesia in goats and sheep Part I. Local
analgesia. Compend. Contin. Ed. Pract. Vet. 1986; 8: S33-S39.

GREEN, C.J. Laboratory animal handbook 8. Animal anesthesia. London: Laboratory

Animals Ltd., 1982.

HALL, L.W. and CLARKE, K.W. Veterinary anesthesia. 9th Ed. Toronto, Ont.: Baillire
Tindall, 1991.

HATCH, R.C., JERNIGAN, A.D., WILSON, R.C., LIPHAM, L.B., BOOTH, N.H.,
CLARK J.D. and BROWN, J. Prompt arousal from fentanyl-droperidol-pentobarbital
anesthesia in dogs: a preliminary study. Can. J. Vet. Res. 1986; 50: 251-258.

HOLMES, D.D. Clinical laboratory animal medicine. Ames, IA: Iowa State University
Press, 1984.

HOLZGREFE, H.H., EVERITT, J.M. and WRIGHT, E.M. Alpha-chloralose as a canine

anesthetic. Lab. Anim. Sci. 1987; 37: 587-595.

HRAPKIEWICZ, K.L., STEIN, S. and SMILER, K.L. A new anesthetic agent for use in
the gerbil. Lab. Anim. Sci. 1989; 39: 338-341.

INGWERSEN, W., ALLEN, D.G., DYSON, D.M., PASCOE, P.J. and O'GRADY, M.R.
Cardiopulmonary effects of a ketamine hydrochloride/acepromazine combination in
healthy cats. Can. J. Vet. Res. 1988; 52: 1-4.

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JERNIGAN, A.D., WILSON, R.C., BOOTH, N.H., HATCH, R.C. and AKBARI, A.
Comparative pharmacokinetics of yohimbine in steers, horses and dogs. Can. J. Vet. Res.
1988; 52: 172-176.

JOHNSON, J.H. Anesthesia, analgesia and euthanasia in reptiles and amphibians. In:
Schaeffer, D.O., Kleinow, K.M. and Krulisch, L., eds. The care and use of amphibians,
reptiles and fish in research. Bethesda, MD: SCAW (Scientists Center for Animal
Welfare), 1992.

JONES, D.M. The sedation and anesthesia of birds and reptiles. Vet. Rec. 1977; 101:
340-342.

KERO, P., THOMASSON, B. and SOPPI, A. Spinal anesthesia in the rabbit. Lab. Anim.
1981; 15: 347-348.

LEVY, D.E., ZWIES, A. and DUFFY, T.E. A mask for delivery of inhalation gases to
small laboratory animals. Lab. Anim. Sci. 1980; 30: 868-870.

LIPMAN, N.S., MARINI, R.P. and ERDMAN, S.E. A comparison of ketamine/xylazine

and ketamine/xylazine/acepromazine anesthesia in the rabbit. Lab. Anim. Sci. 1990; 40:
395-398.

LUDDERS, J.N., MITCHELL, G.S. and SCHAEFER, S.L. Minimum anesthetic dose
and cardiopulmonary dose response for halothane in chickens. Am. J. Vet. Res. 1988; 49:
929-932.

LUMB, W.V. and JONES, E.W. Veterinary anesthesia. Philadelphia, PA: Lea and Febiger,
1984.

LYNCH, S. and LINE, S. Failure of yohimbine to reverse ketamine anesthesia in rhesus

monkeys. Lab. Anim. Sci. 1985; 35: 417-418.

MAHMOUDI, N.N., COLE, D.J. and SHAPIRO, H.M. Insufficient anesthetic potency of
nitrous oxide in the rat. Anesthesiol. 1989; 70: 345-349.

MCLAUGHLIN. S. Anesthesia-Barbiturates. Part I and II. CALAS (Can. Assoc. Lab.

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MOODY, K.D., BOWMAN, T.A. and LANG, C.M. Laboratory management of the ferret
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MORELAND, A.F. and GLASER, C. Evaluation of ketamine, ketamine-xylazine and

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MUIR, W.W. and HUBBELL, J.A.E. Handbook of veterinary anesthesia. Toronto, Ont.:
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MULDER, J.B. and HAUSER, J.J. A closed anesthetic system for small laboratory
animals. Lab. Anim. Sci. 1984; 34: 77-78.

NATIONAL RESEARCH COUNCIL (U.S). Laboratory animal management: Wild birds.

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anesthetic agent in the Mongolian gerbil (Meriones unguiculatus). Lab. Anim. 1983; 17:
324-329.

OLSON, M.E. Problems associated with urethane anesthesia. CALAS (Can. Assoc. Lab.
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OLSON, M.E. A simple anesthetic chamber. Lab. Anim. Sci. 1986b; 36: 703.

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Comparison of ketamine, ketamine and xylazine, droperidol and fentanyl, and sodium
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PADDLEFORD, R.R., ed. Manual of small animal anesthesia. New York, NY: Churchill
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PARKER, J.L. and ADAMS, H.R. The influence of chemical restraining agents on
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RETTIG, B. An inexpensive anesthetic gas-scavenging device that any technician can

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clinical utility of medetomidine/ketamine and xylazine/ketamine combinations for the
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The effects of pentobarbital, fentanyl-droperidol, ketamine-xylazine and ketamine-
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Lab. Anim. Sci. 1989; 39: 411-416.

XII. EUTHANASIA

A. INTRODUCTION

The term "euthanasia," is derived from the Greek terms "eu" for "good" and "thanatos"
for "death" or an easy death (Bennett, Brown, Schofield et al. 1990). However, the term
"euthanize" will be used in this Chapter, rather than the more accurate "euthanatize."
Whichever term is used, the method must be "humane": that is, it must be painless, must
minimize fear and anxiety, be reliable, reproducible, irreversible, simple, safe and rapid.
If possible, it should also be esthetically acceptable for the person carrying out the
procedure, as well as for any observer.

In the 1950s, the term "euthanasia" was rarely heard; euphemisms included "sacrifice,"
"destroy," "put down," or "put to sleep" (Zweighaft, 1990). However, rarely were these
terms prefixed with the word "humane"; nor was it considered necessary. Humane
Slaughter Acts were not promulgated and applied until the late 1950s and early 1960s in
Canada or the USA. Even with the advent of legislation in this regard, many of the meat-
producing species, e.g., fowl, were not included under the Regulations.

In the use of animals in research, teaching, and testing it is essential that the scientific
community take on the mantle of responsibility for applying scientific judgement and
new knowledge to ensure that, when the life of an animal is taken, it is assured of a "good
death." Even with non-consumptive or non-invasive studies using animals, there are
occasions when it is necessary to euthanize the animal (e.g., return of a wild animal to a
hostile habitat) (see also Categories of Invasiveness found elsewhere in this Guide).

In the first edition of this volume of the Guide, the Chapter on euthanasia states: "The
most important criterion of acceptance of a euthanasia method as humane is that it have
an initial depressive action on the Central Nervous System (CNS) to ensure immediate
insensitivity to pain." Although the principle of this criterion remains sound, to the
original wording should be added "to produce rapid unconsciousness and thus assurance
of insensitivity to pain; this must be followed by cardiac and respiratory arrest."

It is important that Russell and Burch's (1959) "Three R" principle noted elsewhere in
this Guide, be equally applied to methods of euthanasia. Refinement of procedures is an
often-neglected area and one which should be addressed in order to ensure that the
criteria for a humane death are in place.

The application of these euthanasia guidelines requires the use of professional judgement,
technical competence, coupled with an understanding of the animal, its behaviour and its

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physiology, as well as an understanding of the environmental and ecological impact, the
sensitivities of other personnel and the concerns of the general public.

B. CRITERIA FOR A HUMANE DEATH

The person applying the method of euthanasia is a most important factor in ensuring that
an animal's death is humane. Regardless of whether the procedure is applied to an
individual animal or to a group, it must always attempt to meet the following criteria:

a) death without signs of panic, pain or distress;

b) minimum time to loss of consciousness, i.e., shortest lag time;

c) reliability and reproducibility;

d) safety for personnel involved;

e) minimal undesirable physiological and psychological effects on the animal;

f) compatibility with the requirement and the purpose of the scientific study;

g) minimal or no emotional effects on the observer and the operator;

h) minimal environmental or ecological impact;

i) simple, inexpensive mechanical equipment which is relatively maintenance free; and;

j) a location remote and separate from the animal rooms.

It is often difficult to recognize evidence of stress when animals are euthanized in the
presence of other animals. Recent information on pheromones provides evidence that
animals can communicate with one another through various types of signals. In certain
experiments with rats, stress induced by experimental treatment may give rise to the
production of signals that affect non-treated animals housed nearby (Duncan and
Petherick, 1991; Beynen, 1992; Short and Van Poznak, 1992).

C. PAIN AND STRESS

The control of animal pain is discussed elsewhere in this Guide (see Control of Animal
Pain in Research, Teaching and Testing) and should be reviewed by those conducting
euthanasia procedures. The literature is growing regarding animal pain (Dawkins, 1980,
1990; Bateson, 1991; Flecknell, 1984; Wall, 1992; Fosse, 1991; Rowsell, 1992). It is
sufficient to note that, over the past 25 years, there has been a revolution in our
understanding of pain mechanisms which was generated by experimental work on
animals. We cannot get inside the head of the animal; thus, an important part of assessing
animal pain is empathy coupled with the ethical concerns.

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Briefly, it is believed that, for pain to be experienced, the cerebral cortex and sub-cortical
structures must be functional. If the cerebral cortex or sub-cortical structures are rendered
non-functional by any method, such as hypoxia, pharmacological depression, electric
shock or concussion, then the feeling of pain is inhibited. Unfortunately, we have no
critical means of determining the adequacy of anesthesia and the assessment of its depth
in animals, whereas a range of different measures have been used for determining the
adequacy of anesthesia in humans (Whelan and Flecknell, 1992). The crude criteria
available to judge unconsciousness in an animal include, for example, the absence of a
blinking reflex, toe pinch reflex and tail reflex. Rarely is an electroencephalogram (EEG)
available showing complete flattening of the EEG as an indication of brain death. This is
the criterion adopted by the American Academy of Neurology as acceptable for
establishing brain death in young children after other clinical criteria, such as deep coma,
failure to breathe spontaneously, and the absence of reflexes occur (Anon., 1987).

In assessing death, it is important to observe that heart action has stopped, thus ensuring a
cessation of blood delivered to the brain, as well as the cessation of respiration. No
animal should be considered dead until reflex movement as well as cardiac and
respiratory movements, have ceased.

If an animal has been given a curare-like preparation, the absence of reflexes should not
be used to indicate unconsciousness and thus insensitivity to pain.

D. MECHANISMS FOR CAUSING DEATH

Death can ensue when the brain is affected by hypoxia, direct or indirect, when there is
direct depression of neurons essential for living physiological functions, or there is
physical disruption of brain activity to produce unconsciousness.

a) In hypoxia, death must be considered painless and free of stress only when
unconsciousness precedes the loss of muscle activity (paralysis). Paralyzing agents (e.g.,
curariform agents such as curare, succinylcholine, gallamine, nicotine sulphate,
magnesium or potassium salts and other neuromuscular blocking agents) must never
be used alone for killing animals (Rowsell, 1990). Following unconsciousness due to
brain hypoxia, some animals exhibit a degree of reflex motor activity.

b) Direct depression of neurons: the depression of neurons of the brain which produces
unconsciousness and then death, is sometimes associated with vocalization and muscle
activity. Death is produced by hypoxemia, direct depression of the respiratory centres in
the CNS, or cardiac arrest.

c) Physical disruption of brain activity produces immediate unconsciousness;

however, there may be marked physical muscular activity due to the depolarization
of the nerve cells. While the movement is esthetically unpleasant, it is not a
manifestation of pain or distress: the animal feels nothing.

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E. METHODS USED FOR EUTHANASIA

1. Physical

Physical methods of euthanasia include stunning, cervical dislocation, electrocution,

pithing, decapitation, shooting, maceration, microwave radiation, and exsanguination.

In the laboratory, physical methods are normally restricted to those animals which are
easily handled, such as small rodents, poultry, large domestic animals, and some
amphibians and reptiles. If the research protocol requires a physical method of euthanasia
because other methods could invalidate the scientific study, the use of such methods must
be justified by the scientist and approved by the Animal Care Committee (ACC). Prior
sedation or tranquillization should take place whenever possible.

Decisions to use physical methods for euthanasia must be based on professional

judgement and be undertaken only by experienced individuals. Acquiring (or re-
acquiring) the skills to use physical methods of euthanasia may be accomplished by
practising the techniques on dead animals, preferably those recently killed, and be subject
to close scrutiny by those experienced in the methodology.

a) Stunning is sometimes used with small laboratory rodents. The blow must be
delivered to the central skull bones with sufficient force to produce massive cerebral
hemorrhage and thus immediate depression of the CNS, producing rapid
unconsciousness. This technique should not be undertaken in the presence of casual
observers or the uninformed, for it is esthetically unpleasant. However, when properly
applied, the animal is immediately rendered unconscious and thus insensitive to pain.
Subsequent to stunning, the animal's major blood vessels should be cut, and the chest and
the heart opened.

b) Cervical dislocation is suitable for poultry, mice, immature rats or rabbits, or similar
small species. The technique consists of separation of the skull and the brain from the
spinal cord by pressure applied posterior to the base of the skull (Clifford, 1984). When
the separation of the cord occurs, CNS stimulation of respiration and heart beat is
interrupted, leading to death. The supply of blood to the brain continues to nourish it
because the carotid arteries and jugular veins are intact; however, the blood will rapidly
be depleted of oxygen and there will be an increase in carbon dioxide after respiration
ceases, leading to brain disfunction.

Studies have demonstrated that the EEG flattens and the blinking reflex disappears
immediately after the spinal cord separates, thus indicating that the animal is not sensitive
to pain (Allred and Berntson, 1986; Rowsell, 1990; Derr, 1991). In addition, the severed
spinal cord does not deliver a painful stimuli from areas posterior to the separation; thus,
with the separation of the spinal cord from the brain, painful stimuli cannot be perceived.
However, significant muscular movements may take place.

The Report of the AVMA Panel on Euthanasia (AVMA, 1993) differs from its 1986

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predecessor in that it notes that data suggest that electrical activity in the brain persists for
13 seconds following cervical dislocation (Vanderwolf, Buzsaki, Cain et al. 1988). In
addition to mice, it lists as suitable subjects immature rats weighing less than 200 g,
rabbits weighing less than one kg, poultry and other small birds. It also notes that: "In
heavier rats and rabbits, the greater muscle mass in the cervical region makes manual
cervical dislocation physically more difficult; accordingly, it should be performed only
with mechanical dislocators or by individuals who have demonstrated proficiency in
euthanatizing heavier animals." The need for proper training is stressed.

c) Decapitation with guillotine is used primarily to euthanize rodents and small rabbits.
Used alone, it provides a means of ensuring that tissues and body fluids are chemically
uncontaminated as well as providing a means of obtaining an anatomically intact brain
and brain tissues for further study.

After consulting the literature (Vanderwolf, Buzsaki, Cain et al. 1988; Derr, 1991;
Mikeska and Klemm, 1975), the Canadian Council on Animal Care (CCAC) concurs with
the Report of the AVMA Panel on Euthanasia (AVMA, 1993) that pre-sedation before
decapitation or cervical dislocation is not necessary. However, the use of cervical
dislocation and decapitation with guillotine as euthanasia methods must be
scientifically defended by the investigator and approved by the ACC. Well-designed
and easily operated guillotines are available from commercial sources. Guillotines should
not be used by personnel who have not been properly trained in the methodology and
how to properly handle the animal. Decapitation as a means of euthanizing amphibia and
reptiles is not recommended (AVMA, 1986; Cooper, Ewbank, Platt et al. 1989).

d) Pithing, which is used to euthanize frogs and turtles by destroying the brain after the
frog has been anesthetized, requires considerable skill. A sharp, pointed probe is inserted
through the skin between the skull and the atlas. It is then pushed forward through the
foramen magna into the cranial cavity, using a twisting motion. The technique should be
attempted only after acquiring knowledge of anatomy using skeletons, and after a period
of training including practice on dead animals. This method can cause pain and suffering
if the proper regions of the brain are not completely destroyed.

e) Shooting by penetrating captive bolt pistol has been used primarily for pre-slaughter
stunning of food animals. To a lesser extent, it is used in emergency situations such as
roadside injuries or other similar events where no other method, or the expertise
necessary to apply it, is available.

Captive bolt pistols are also used in emergency situations in slaughter of horses.
However, because of their behavioural/physical response of rearing on their hind legs and
falling backwards, unless proper restraints are applied, there is a danger to the operator. It
is essential that the captive bolt pistol be withdrawn from the animal's head
instantaneously. An extension handle is attached to the pistol to allow the operator to put
one or both hands in front of the horse's eyes, thus reducing head tossing. This specially
designed captive bolt pistol for killing horses requires knowledge of the placement
needed in order to penetrate the deep structures of the brain (Watts, 1976).

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Britains' Royal SPCA, following the use of the captive bolt pistol in the abattoir,
developed a smaller, hand-held model for emergency euthanasia of injured dogs and cats
which was used by some SPCAs and humane societies (UFAW, 1968). In Canada,
however, it was considered esthetically repugnant by the general public and therefore its
use was limited to those occasions when the injured animal could be removed from
public view (UFAW, 1988).

More recently, a captive bolt pistol was developed for killing rabbits and goats (Accles
and Shelvoke Ltd., Aston, Birmingham, Eng. B64QD). With this device, the clinical
evidence, including the loss of corneal reflex and organized EEG activity, suggested that
loss of consciousness and cerebral death occurred almost immediately (Dennis, Dong,
Weisbrod et al. 1988).

Only commercially produced penetrating or captive bolt pistols should be used. Proper
utilization is essential. Considerable technical competence and precise knowledge of the
animal's anatomy are necessary; therefore, this method should be used only by an
experienced operator. Exsanguination must follow unconsciousness.

f) Percussion stunning is carried out by means of a non-penetrating form of captive bolt

stunning device with mushroom-shaped tip. The effects on brain function will depend on
the speed of the bolt on impact, its proper positioning, and the thickness of the cranial
bone. Unfortunately, the minimum speeds necessary for effective percussions have not
yet been determined. The value of this method is that it can be used to replace some ritual
slaughtering methods. Percussion stunning has been used as an alternative method to the
penetrating captive bolt by some Canadian abattoirs. Unfortunately, the instruments are
more subject to breakdown and malfunctioning than the standard penetrating captive bolt.
More recently, a percussion-type instrument has been developed for use in animal
control, particularly for dogs. Its feasibility and its humaneness as a technique need to be
established in the field operations.

Captive bolt pistols, including percussion stunning devices, require permits for their
possession in Canada.

g) Shooting is an effective means of humanely destroying animals in the field. Only

experts should carry out this procedure. The subject must be shot at close range and the
bullet must strike the brain so as to render the animal immediately insensitive to pain; 12-
bore or 20-bore shotguns, or 22-calibre rifles or revolvers may be employed, depending
upon the species and size of the animal to be killed. Whether or not prior sedation should
be used is debatable (Ebedes, 1988). The use of shooting by firearms in a laboratory
setting is prohibited. In such settings, there are always other available methods of
euthanasia and individuals with appropriate expertise.

There are advantages of shooting as a euthanasia method in the field, for unconsciousness
is instantaneous if the bullet destroys a significant portion of the brain involving
particularly the vital centres. In the field, this may be the only possible method to render

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the animal immediately unconscious and to produce death. The need for expertise of the
marksman cannot be overemphasized. For use in emergency situations, Guidelines for
Euthanasia of Domestic Animals by Firearms have been prepared by the Animal Welfare
Committee of the Canadian Veterinary Medical Association (CVMA) (Longair, Finley,
Laniel et al. 1991).

h) Electrocution, used mainly to kill domestic animals (Eikelenboom, 1983) is rarely

used in the laboratory setting. If electric shock is used, it must be delivered in two phases:
the first electric shock passes through the brain, stunning the animal; the second,
delivered a fraction of a second later, produces a fibrillation of the heart, killing the
animal. Violent extension of the extremities, part of the seizure induced by electrocution,
is esthetically unpleasant.

i) Microwave radiation is a relatively new technique employed mainly by neuroscientists

who wish to maintain the anatomic, enzymatic and physiological chemical composition
of the animal's brain in an unaltered state (Stavinoha, 1983; Ikarashi, Maruyama and
Stavinoha, 1984). Microwave radiation must be delivered specifically to the brain;
therefore, standard household microwave ovens must not be used (Stavinoha, Frazer and
Modak, 1977); only instruments which have been designed specifically for this
purpose and have the appropriate power and microwave distribution may be
utilized.

j) High altitude decompression is considered unacceptable by the CCAC. At one time,

it was used by some animal control agencies and humane societies for killing unwanted
dogs and cats, but is now not recommended by them (White, 1984). When animals have
upper respiratory problems or gastrointestinal upsets, the gases in these areas, as well as
in the sinuses, expand and cannot be vented, thus producing significant pain and distress
(White, 1984).

k) Exsanguination (depriving an animal of blood) is only acceptable as a euthanasia

procedure if the animal is first rendered unconscious by physical means such as stunning,
or pharmacological means, e.g., injection of an anesthetic (Gregory and Wotton, 1984).
The use of this method must be scientifically justified and approved by an ACC, who
must have established the technical competence of the person who will conduct the
technique.

l) Maceration requires the application of severe restrictions on the size and age of the
animals that may be subjected to this process (Ewbank, 1987). For example, it is used to
kill newborn mice, and in some poultry operations for killing surplus one-day-old male
chicks, where it has been deemed acceptable by Agriculture Canada. Equipment has been
specifically designed for this purpose and, although esthetically unpleasant to consider as
well as to observe, the method produces instantaneous unconsciousness and death when
used properly.

m) High pressure water jet has recently been proposed for stunning slaughter pigs
(Schatzmann, Leuenberger, Fuchs et al. 1991). However, its use has not been reviewed

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by other experts qualified to evaluate humane slaughter techniques.

2. Non-inhalant Pharmacologic Agents

The majority of injectable drugs used as anesthetic agents are acceptable for euthanasia if
an adequate overdose is given. The preferred route is intravenous (IV), and should be
accompanied by adequate restraint, making the animal as comfortable as possible with
minimum distress or anxiety. Pre-sedation or tranquillization may be necessary for wild,
feral or fearful animals not accustomed to restraint.

If the animal is too small to receive intravenous injections, or if anatomically suitable

veins are not visible or apparent, e.g., in small rodents and guinea pigs, the intraperitoneal
(IP) injection of a non-irritating overdose of a pharmacological agent is acceptable.

With most of the available injectable anesthetic agents, the amount to be injected is too
large to use the intramuscular, subcutaneous, intrathoracic, intrapulmonary, and
intrathecal sites. Administration by a route other than intravenous in most cases results in
a delayed onset of the anesthetic effect of the drug. Under these circumstances, it is
essential that the animal be placed in a cage or enclosure to ensure prevention of injuries
through stumbling and or falling, and in order to make the animal more comfortable and
facilitate onset of the overdose of anesthetic.

a) Barbituric acid derivatives (barbiturates) used as anesthetics are effective in

producing euthanasia when given as an overdose. The pharmacological action of these
drugs is to depress the CNS, starting with the cerebral cortex, and progressing through the
stages of anesthesia to produce unconsciousness. With an overdose, deep anesthesia
develops, followed by apnea as the respiratory centre is depressed, followed by cardiac
arrest and death. Some of the combination barbiturate derivatives have a cardiotoxic
effect; however, this is of no consequence because the animal dies before such an effect
histologically manifests itself.

Barbituric acids are controlled substances under regulation by the Bureau of Dangerous
Drugs, Health and Welfare Canada. As controlled substances, they must be stored in
locked cabinets and a written log maintained of date and amount used and the purpose of
use. Euthanasia agents containing barbituric acid derivatives are often coloured to make
them clearly identifiable. The amount used for euthanasia should be in accordance with
the manufacturer's direction. The abuse or misuse, either accidental or deliberate, of such
substances creates a significant risk and associated legal liability. Those possessing such
combinations for euthanasia must provide adequate security.

b) T-61 is manufactured by Hoechst-Roussel Canada Ltd. (4045 Cote Vertu, Montreal,

Quebec, H4R 2E8). It contains a local anesthetic (tetracaine Hcl), a strong hypnotic agent
which depresses the CNS causing unconsciousness (brain death), as well as a curariform
drug which has a paralytic effect on the respiratory centre and a relaxing effect on
skeletal muscles (Rowsell, 1979). A recent study demonstrated that induction of muscle
paralysis and unconsciousness occur simultaneously (Hellebrekers, Baumans, Bertens et

269
al. 1990). These authors concluded that the muscular activity and vocal response seen in
some dogs was not a conscious response.

T-61 should be administered intravenously at the dose and administration rate directed by
the manufacturer. If instructions are not followed, it is possible for T-61 to produce an
excitatory phase and vocalization. It is not registered or restricted by the Bureau of
Dangerous Drugs and may be used by non-medical, technical personnel. However, it
must be ordered by a veterinarian and be shipped directly to the veterinary clinic involved
(Clarke, 1990). Its availability in other countries has been affected because of criticism
levelled primarily as a result of failure to follow the manufacturer's direction for the
dosage and the rate of injection. Although this is not a restricted drug, the same
guidelines concerning its safekeeping apply as to any of the barbituric acid derivatives
and anesthetics, for its use has been abused (Smith and Lewis, 1989).

c) Chloral hydrate is a dissociative anesthetic and produces no loss of the corneal or

blinking reflex. Difficulties include slow onset of action, restraint difficulties, and the
amount that must be administered. Death is due to hypoxemia caused by progressive
depression of the respiratory centre. It may be preceded by gasping, muscle spasms, and
vocalization, causing difficulties in comfortably restraining the animal being euthanized.
Therefore, while it should not be used for dogs and cats, it is acceptable for
intravenous use in large animals and is an effective euthanasia agent for poultry.

A combination of chloral hydrate, magnesium sulphate, and sodium pentobarbital,

administered intravenously to large domestic animals as an overdose, is an acceptable
method of euthanasia.

d) Ketamine hydrochloride, also a dissociative anesthetic, cannot be recommended for

euthanasia because it is difficult to assess what constitutes an overdose.

e) Magnesium sulphate alone is a neuromuscular blocking agent (Hevner and de Johng,

1973); however, it does not depress the CNS. Administration of magnesium sulphate
should only be used in combination with barbituric acid derivatives and administered
only by the IV route. The IP route is not acceptable because of the irritating nature of a
saturated solution.

3. Inhalant Anesthetics

An overdose of inhalant anesthetics such as ether, halothane, methoxyflurane, isoflurane,

and enflurane fulfils the principle of a humane death. Their use, however, poses a risk to
human subjects who may be exposed to their vapours; thus, they are considered an
occupational hazard (see Anesthesia, as well as Occupational Health and Safety).
Chambers are available commercially to expose animals to such anesthetic gases in order
to either produce anesthesia or, by overexposure, to produce euthanasia. Scavenging
systems to remove excess gases are readily adapted to such enclosures. Additionally,
anesthetic masks can be prepared to fit even small rodents. The vapours are inhaled until
respiration ceases. The animal is then checked to ensure that it is dead.

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The soaking of gauze with inhalant anesthetics and then placing it in a container with the
animal(s) to be euthanized may be used only if there are no other methods of delivery of
the anesthetic gases. The fact that inhalant anesthetics are liquid makes it essential that
animals be exposed to vapours only, as the liquid form is a topical irritant. The delivery
system should provide sufficient oxygen with the anesthetic vapour to ensure
unconsciousness precedes hypoxia.

a) In the past, chloroform and ether have been commonly used as anesthetics or, when
exposure to their vapours is of sufficient concentration and duration, to produce
euthanasia. However, chloroform is no longer recommended because of its
carcinogenic, hepatotoxic and nephrotoxic potential. Ether is a flammable and
explosive agent, and should never be used in the presence of flame or where electrical
equipment is not protected and shock resistant.

b) Nitrous oxide is of value as an agent for euthanasia only in combination with other
volatile inhalant anesthetics. It is a combustible agent, but is non-flammable and non-
explosive. With the exception of ether, most inhalant anesthetic agents are expensive and
require special anesthetic delivery mechanisms; thus, their use as euthanasia agents is
limited to species where venipuncture is found to be too difficult or impossible.
Additionally, use of inhalant anesthetics for euthanasia of large animals is expensive
because of the amounts that must be used.

4. Non-anesthetic Gases

Non-anesthetic gases include carbon monoxide, carbon dioxide, nitrogen, argon and
cyanide.

a) Carbon monoxide, even in low concentrations, can harm other animals and humans
exposed to its fumes. A colourless, odourless gas, it is difficult to detect. Carbon
monoxide obtained from the exhaust of a gasoline combustion engine contains impurities
and thus can produce irritation and discomfort. Therefore, delivery of an irritant-free
carbon monoxide is mandatory if this is the chosen method. Carbon monoxide is rarely
used now to destroy unwanted dogs and cats by animal control agencies and humane
societies; however, it continues to be used for some of the fur-bearing species. In the
laboratory, because of the safety problems associated with its delivery, carbon monoxide
is not recommended as a euthanasia agent (Chalifoux and Dallaire, 1983).

b) Nitrogen and argon are inert gases which are both colourless and odourless, non-
flammable and non-explosive. They are considered as having a minimal impact on the
environment or the atmosphere. Both are used in a closed chamber through a process
called "flushing" in which the passage of these gases reduces oxygen levels to a
maximum of 1.5%. At such low levels of oxygen, the animal will collapse, and death will
be produced by hypoxemia. However, dogs, cats, and rabbits may vocalize at the 1.5%
level of oxygen, as well as show increased muscular activity and struggling. Nitrogen and
argon do not produce a narcosis prior to the onset of hypoxia which in itself will lead to

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unconsciousness followed by death resulting from paralysis of the respiratory centre in
the anoxic brain. Carbon dioxide, on the other hand, can induce a narcosis because of its
physiological effect on the CNS (Herin, Hall and Fitch, 1978).

Quine, Buckingham and Strunin (1988) found that use of acepromazine as a tranquillizer
before placing dogs in a nitrogen euthanasia flushing chamber produced longer survival
times. However, it was not recorded whether or not these treated dogs showed the same
amount of hyperventilation prior to or following the loss of consciousness; yelping,
gasping, convulsions and muscular tremors usually accompany this process. Chalifoux
and Dallaire (1983), however, concluded that premedication with tranquillizers improved
the humaneness of carbon monoxide euthanasia.

c) Argon gas is more dense than air and thus tends to remain in the lower layers;
however, both nitrogen and argon have no analgesic or anesthetic properties.

d) Carbon dioxide (CO2) is frequently used to kill rodents and birds in the laboratory.
Although it is a component of room air, pure CO2 is heavier than air and practically
odourless. It concentrates in the lower portion of the euthanasia chamber, and rats and
other burrowing animals will tend to keep their noses in the lower zone containing
adequate concentrations of the gas.

Its use as a euthanasia agent is dependent (for its humaneness) on whether or not it is in
sufficient concentration to produce a narcosis. Maintenance at the correct level requires
some manipulation to physiologically achieve, but will produce a narcosis and if oxygen
levels are not increased, will lead to death. Carbon dioxide also stimulates the respiratory
centre in the brain and in low concentrations of up to 10% of inspired gas is considered a
potent respiratory stimulant causing a tenfold increase in the ventilation rate and a feeling
of profound respiratory distress. The stimulation of the respiratory centre produces
hyperventilation, thus critically affecting the onset of CO2 narcosis. At approximately
40%, the CO2 induces anesthesia which is slow in onset and accompanied by involuntary
excitement. Eventually, there is apnea, a fall in blood pressure, and death (Ontario
Ministry of Agriculture and Food Memo to Pound Operators and Veterinarians in
Ontario, August 12, 1987). Britt (1986) found that the slow induction of narcosis is
preferable, as using a precharged chamber, or passing CO2 into the chamber too rapidly,
can produce obvious signs of distress in the animals. However, he concluded that "neither
(slow nor swift) method was found to be stress-free, so no recommendation can be a
counsel of excellence."

McArthur (1976) reported on a method for delivering a carbon dioxide euthanasia of

small animals, including cats, puppies, mice, rats, gerbils, guinea pigs and hamsters. In
this study, unconsciousness was produced without distress to the animals, and occurred
within a minute when levels of oxygen were maintained at 31 to 33%, with CO2 ranging
from 56 to 63%. Unconsciousness was used to describe animals that had collapsed and
showed complete relaxation. Deep surgical anesthesia was produced by leaving the
animals in this environment for a three-minute exposure period. Once deep surgical
anesthesia was reached, the oxygen supply was turned off and CO2 filled the chamber.

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Carbon dioxide may be purchased in cylinders or in the solid state, as dry ice. It is
relatively inexpensive, non-inflammable, non-explosive and essentially non-hazardous. It
presents no danger to operators or attendants when used with properly designed
equipment in an adequately ventilated space.

The essential criteria for producing a CO2 narcosis is to maintain the oxygen level close
to, or slightly below normal air levels, and increase the percentage of carbon dioxide in
the air. It is important to understand that newborn animals, having existed in an
environment with low oxygen levels prior to parturition, require higher levels of carbon
dioxide in order to produce a humane death. Therefore, newborn animals should not be
removed from chambers charged with carbon dioxide for approximately one-half hour
after all movement has ceased.

Carbon dioxide does not accumulate in tissues; thus, there is no residue in food-
producing animals. Neither does it appear to cause any distortion of the cells, which
appear normal under microscopic examination.

Dogs, cats and other larger animals with an investigative behaviour will often extend
their heads above the zone of effective CO2 concentrations and thus be exposed to levels
that excite rather than depress the CNS; thus, some will hyperventilate, struggle, stagger,
and fall. The important element is to maintain an even distribution of CO2 in the
euthanasia chamber; this requires additional equipment.

Carbon dioxide has proven to be non-effective in killing diving mammals that have
adapted to a relatively anerobic (oxygen-free) environment; 100% CO2 is required to kill
mink. Carbon dioxide is also used for pre-slaughter stunning in swine (Gregory, Moss
and Leeson, 1987).

e) Potassium cyanide is a very potent paralyzing agent of the respiratory centre. It is one
of the most rapidly acting poisons. Death appears to be almost instantaneous and
irreversible through the production of rapid anoxia with CNS depression. However, death
by exposure to cyanide gas is not considered humane because convulsions or seizures
occur prior to death. As well, because of the extreme danger associated with its use,
cyanide is not recommended as a method in the laboratory.

F. SPECIFIC SPECIES

In addition to the euthanasia methods for common laboratory species noted above, the
following apply to:

1. Amphibians, Fishes and Reptiles

The most commonly used method for euthanizing amphibians, fish, and reptiles is by
stunning, using the methodology previously outlined for vertebrates and terrestrial
mammals. This can be followed by decapitation or crushing of the skull.

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Sodium pentobarbital and barbituric acid derivatives may be used intravenously or
introduced directly into the abdominal or pleuroperitoneal cavity of most cold blooded
animals, if their anatomy permits. Tricaine methanesulfonate (MS-222) may be
administered by a variety of routes to induce euthanasia. For aquatic mammals, as well as
amphibians and fish, this material may be placed in the water. Alternatively, in the case of
large fish, a gill cover is placed over the gills and concentrated MS-222 is flushed over
the gills. Benzocaine hydrochloride is a compound similar to MS-222 and may be used as
a bath or in a recirculation system for the euthanasia of fish.

It is worth noting that many experts in the study of amphibians and reptiles approve of
the use of cold for the anesthesia of these animals. Subsequent freezing or decapitation is
not considered to be painful; unfortunately, there is no evidence that cooling to 4oC will
lessen the pain threshold (Cooper, Ewbank, Platt et al. 1989). Cooper (1986) states that
hypothermia would have to be instantaneous if it were to be painless.

Inhalant anesthetic agents, such as halothane, methoxyflorane, etc., may be used to kill
reptiles and amphibia either in a chamber or by a suitably adapted face mask. Special
attention must be paid to ensure they are dead following application of the method.

Carbon dioxide is not suitable for all species and concentrations must be maintained at a
high level. Many semi-aquatic and terrestrial mammals, as well as reptiles and amphibia,
are accustomed to living without oxygen and have a tolerance for hypoxia; thus, they
have exaggerated anaerobic capabilities (Hochachka, 1980) (see also Chapter 22, Wild
Vertebrates in the Field and in Laboratory in Vol. 2 of this Guide [CCAC, 1984]).

Decapitation as a means of killing amphibians and reptiles is unacceptable because of the

differences in their physiology (Cooper, Ewbank, Platt et al. 1989; AVMA, 1986).

2. Domestic Animals Killed for Food

Although this chapter is concerned primarily with experimental animals, there are times
when farm animals in production studies or, in some cases, in biomedical studies, will
ultimately be killed for their food value. Although we do not apply the term "euthanasia"
to the killing of such animals, the principles enunciated in the foregoing and in other
documents apply: it is essential that humane treatment of animals be provided. In North
America, the term "humane slaughter" is commonly used; however, in the United
Kingdom, the term used is "pre-slaughter stunning" (Cockram and Corley, 1991;
Ewbank, Parker and Mason, 1992; Knowles and Warriss, 1992). Pre-slaughter handling,
the amount of time the animal is brought to the area to be killed and when it is stunned,
and the proper design of restraint equipment affect the level of stress during handling,
stunning and slaughter (Grandin, 1992a; 1992b). A comparison has been made between
dislocation and percussion in chickens (Gregory and Wotton, 1990).

All killing of food animals must comply with federal legislation (Agriculture Canada's
Meat Inspection Act S.C., 1985, C17 [May 1985 Part 2, Item 39, for 851078S14]; the

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federal Health of Animals Act [C-66 June, 1990, rev. March 1992]; 38-39 Elizabeth II
Chapter 21]). As well, slaughter is discussed in Agriculture Canada Codes of Practice for
the various livestock species (Agriculture Canada, 1771/E, 1984; 1821/E, 1988b; 1757/E,
1989b; 1833/E, 1990; 1870/E, 1992) as well as mink and foxes (Agriculture Canada,
1819/E, 1988a and 1831/E, 1989a) and provincial legislation (see Legislation). Provincial
legislation on humane slaughter, where it exists, as well as municipal and local bylaws
must also be followed.

3. Fur-bearing Animals

Mink, foxes, chinchillas, nutria and opossums are often maintained in research facilities.
The methods listed above can be applied to the killing of these species. Ranched mink are
commonly killed by the use of carbon monoxide, carbon dioxide and nitrogen (Hansen,
Creutzberg and Simmonson, 1991) or electrical stunning followed by cervical
dislocation.

G. TISSUE EFFECTS OF EUTHANASIA METHODS

Tissue effects of euthanasia methods may be either direct or indirect. They may affect
only components in the intravascular compartment or they may affect fixed tissues, thus
influencing histological or electronmicroscopic findings. In the majority of cases, death is
so rapid that even electronmicroscopic changes are non-existent or minuscule. In most
instances, the concern of the investigator that demonstrable histocytotoxic changes may
occur is unfounded.

1. Direct Effects

In general, the direct effects of the euthanizing method are subtle or lacking, particularly
with the non-inhalant pharmacological agents. The changes produced by methods which
cause anoxia depend upon the rapidity of the induction of the anoxic state, and occur as
the result of changes in blood gases. For example, pulmonary congestion and edema may
be observed on gross observation in the anoxic state; however, the degree is dependent
upon rapidity of death.

Lamellar bodies in Purkinje cells of the cerebellum have been observed in some hypoxic
anesthetized dogs subjected to rapid decompression (Bowman, Cooke and Carry, 1969).
The knowledge of the sequential series of morphological and biochemical events leading
to hypoxic injury to neurons and glial cells has not been adequately studied (Kim, 1975).
There is nothing to suggest that hypoxic changes produced by carbon dioxide make
tissues unsuitable for routine examination of the respiratory tract (Fawell, Thompson and
Cooke, 1972).

Barbiturates ionize on injection into the intravascular compartment. The degree of

ionization will depend upon the dissociation constant of the drug and the Ph of the blood.
Cell penetration can occur only with the undissociated drug. After cell penetration takes
place, dissociation again occurs and the binding of the drug to intracellular organelles

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will take place. Tissue changes due to cellular penetration and intracellular binding
of barbiturates have not been described. At the same time as the barbiturates bind with
plasma proteins forming an equilibrium of bound and unbound drug in circulating blood,
splenic dilation with sequestration of red blood cells produce an enlarged and grossly
blue-black spleen (Lumb, 1974).

2. Indirect Effects

The major indirect effects are due to tissue hypoxia brought on by the death of the
animal. Thus, it is important that tissues for histological and electronmicroscopy
examination be prepared as rapidly as possible following unconsciousness and death of
the animal.

The tissue requirements for oxygen vary widely. The damages to the neurons of the CNS
occur most rapidly, but are dependent upon the degree of tissue hypoxia and time elapsed
after death before tissue preparation. Hypoxia changes to neurons require
electronmicroscopic examination.

In those tissues whose requirements for oxygen are not as great as the neurons (e.g.,
osteocytes, chondrocytes of bone and cartilage, or other less oxygen-sensitive tissues)
changes may be difficult to detect even with electronmicroscopy.

Proper handling of the animal prior to death, with immediate processing of tissues after
death, are important to obtain optimal electronmicrographs with minimal changes.

H. EFFECT ON OBSERVERS

We must recognize that a significant number of people experience emotional uneasiness,

physical and psychological discomfort or distress when animals are killed, even if a
humane method is applied. For example, the 1987 Malouf Commission Report on Seals
and Sealing stated that, although the method used for killing seals (striking the animal's
head with a hakapik) was proven to be humane by numerous veterinary pathologists, the
general public considered the act repugnant. For over a decade, the whitecoat seal on the
ice on the Gulf of St. Lawrence or at the Front of St. Anthony, Newfoundland, was the
only animal that the public had seen being killed in media presentations.

Inexperienced observers might misinterpret any movement, vocalization, or reflex

reactions as indicators of pain and distress. Therefore, it is preferable that euthanasia
methods, in addition to providing a humane death, minimize or eliminate such
involuntary movements.

Over the past several years, recognition has been given to the effect of euthanasia on
those required to carry it out (e.g., humane society workers, laboratory animal
veterinarians, animal caretakers and others whose jobs involve euthanasia) (Rollin, 1986;
Rollin and Kesel, 1990; Grier and Colvin, 1990). Arluke (1992) stated that,
understandably, uneasiness was particularly noticeable among newcomers; with seasoned

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workers, it was most common among animal caretakers; it occurred among technicians,
but was relatively rare among veterinarians and scientists. Owens, Davis and Smith
(1981) have noted that those who must euthanize animals have developed ways and
means of dealing with their emotions by avoiding unnecessary contact with the animals
or by believing that, by being killed, the animal is being spared additional suffering.

Workers can (and will) accept that, when an animal is showing pain and distress that
cannot be relieved, it must be killed. In this, their attitudes differ little from those of an
owner of a companion animal who must face similar decisions. Like the pet owner, the
employee can feel grief; therefore, it is important that strong communications be
established with all staff members, with a willingness to listen and to provide support for
those who may be feeling distressed or disturbed. It is important that such "feelings" not
be suppressed in the research setting. Employees should be aware that, regardless of the
purpose of the experimental study, at an established time and date, the animal must be
humanely destroyed. Seminars or workshops to help staff cope with animal death may be
useful.

I. EUTHANASIA STATEMENTS--OTHER AGENCIES

As noted, the Report of the AVMA Panel on Euthanasia has recently been published
(AVMA, 1993). Euthanasia has been discussed by the U.S. National Research Council
(NRC, 1991). Reference to euthanasia may also be found in the U.K.'s Universities
Federation for Animal Welfare (UFAW) Report on Euthanasia of Unwanted, Injured or
Diseased Animals for Education or Scientific Purposes (UFAW, 1986), its Humane
Slaughter of Animals for Food (UFAW, 1987) and Britain's Animals (Scientific
Procedures) Act, 1986 (Balls, 1986; McKie, 1986; Fisher, 1990). The Australian Council
for the Care of Animals in Research and Teaching has prepared a bibliography which
includes a number of references to euthanasia (ACCART, 1991). Attention is also drawn
to Field Research Guidelines (Orlans, 1988) which lists a number of guidelines (ASM,
1987; AOU, 1988; ASIH, HL, SSAR, 1987; ASIH, AFS, AIFR, 1987; Zwart, deVries and
Cooper, 1989).

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Chapter XIII - The Use of Animals in Psychology

XIII. THE USE OF ANIMALS IN PSYCHOLOGY

The focus of psychology is the organization of behaviour. It is concerned with processes

that control and direct adaptive and maladaptive activity, and so the range of phenomena
studied is broad, and the paradigms and research settings employed are varied. The study
of behaviour may incorporate physiology, pharmacology, ethology, or even sociology,
and for that reason the distinction between psychology and closely related disciplines is
often blurred. Through the history of the discipline, extending back, for example, to
nineteenth century work on reflex organization, through Pavlov's fundamental discoveries
about conditioning, and more recently to the identification of motivational and reward
systems through electrical stimulation of the brain, animal behaviour has played a central
role in research and conceptualization. Although there are clearly recognized limitations
to the use of animals in research (e.g., no access to verbal report), it does permit a control
of hereditary and experiential variables that could seldom be achieved in other ways.
Furthermore, animal research places humans in an evolutionary context and makes

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possible a comparative and biological perspective on human behaviour.

Basic animal research in psychology has played a significant role in advancing our
understanding of processes of learning, memory, perception, motivation, and emotion,
and of behavioural adaptations of individuals and species to their environments. While
much of this work has addressed theoretical issues, it can have direct implications for
contemporary applied problems. Examples of such problems that affect animals directly
include captive and domestic animal care, non-lethal means of predator control, and the
reintroduction of endangered species to their former habitats. Examples of problems that
have a direct effect on human welfare include the control of depression, phobias, pain,
addiction, and the pathological effects of stress and anxiety.

Although it may be convenient to dichotomize research activities as basic and applied, it

is important to recognize that there is a continuum and that it is difficult to know a priori
where along that continuum the implications of some research program will fall.
Applications sometimes follow quite directly from basic research, and fundamental
discoveries often arise in applied research. It is also important to appreciate, as Hebb
(1966) has pointed out: "Before we can have applied science, we must have a science to
apply." This is a position shared by many researchers who, while cognizant of applied
implications, orient their work toward the elucidation of issues that are basically
scientific and theoretical in nature.

Since psychological research approaches a wide range of topics with a diversity of

methodologies, it is not surprising that animal research in psychology may not be well
understood outside the discipline. The following points are directed at research strategies
and general experimental procedures that have apparently been sources of
misunderstanding:

1. Many problems studied by psychologists deal with the understanding and control of
psychopathology, such as depression, phobias, psychosomatic disorders, psychoses,
hyperactivity and learning disabilities, obesity and addiction. Many aspects of these
problems cannot be studied satisfactorily in human patients because of the difficulties
associated with non-experimental paradigms in determining the causal relationship
among variables. In other words, when one studies patients with a form of
psychopathology, all one can establish is that some variable, X, is correlated with the
pathology, P. Yet an understanding and control of the problem requires knowing more
than correlations. It is necessary to establish whether that variable X in some sense causes
or is an antecedent of the pathology P, whether the pathology P is an antecedent for the
change in variable X, or whether the two variables are related only indirectly and non-
causally through their relationship to some common underlying variables yet to be
determined. Seldom is it practicable to study these kinds of important issues in human
patients using the necessary experimental (as opposed to correlational) research designs.
One alternative and productive research approach has been to use animal models. In the
present context, such models refer to "the production, under controlled conditions, of
phenomena analogous to naturally occurring disorders."

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A more extensive discussion of the concept of animal models in psychology may be
found in the literature (Abramson and Seligman, 1977).

Using such models, it becomes possible to conduct experimental studies involving the
active manipulation of variables, and this permits clarification of the relationships among
variables. It is important to recognize, however, that a model is simply that, and requires
validation through a detailed study of its essential features and an analysis of its
similarities to the psychopathology in question.

2. In addition to their use in applied problems, animal models play an important role in
the development of fundamental behaviour theory. Research may study, for example, the
behaviour of animals pressing a lever in an isolated chamber, in order to examine the way
in which the frequency or patterning of lever presses is controlled by the schedule of food
reward or reinforcement. No one is terribly interested in lever pressing as a behaviour in
itself; however, this simple and easily quantified behaviour can be viewed as a model or
analogue of more complex forms of behaviour. The assumption is that if one can
understand the basic principles that control a simple behaviour, one will have at least a
place to start in developing principles that govern more complexly organized behaviour
systems.

Similarly, when researchers use electric shock as a means of producing stress or of

motivating animals to escape or avoid, they are fully recognizant that electric shock does
not normally occur in nature. They do assume, however, that this particular easily
controlled aversive event can serve as a model or analogue of other unpleasant events that
do occur naturally and affect behaviour. This use of models in psychology is often
misunderstood by those outside the discipline. It is important that they note that questions
of the validity of the assumptions and the adequacy and generalizability of a model
cannot be answered a priori and are more productively approached through empirical*
research than logical argument.

* It should be noted that the word empirical has two quite different, but equally correct
meanings of which the reader should be aware. The first refers to work that is based on a
systematic observation and the application of scientific principles in methods. It is in this
sense that the word is normally used in psychology (although there also occurs the word
empiricism, which refers to a philosophy of knowledge). The second meaning of
empirical is found in the medical sphere, and refers to work that "relies on or is based on
practical experience without reference to scientific principles as, an empirical remedy". It
is related to the noun, empiric, which refers to one "ignorant of scientific principles" and
who "lacks regular training and proper qualifications" (Webster's New World Dictionary,
Second College Edition, 1970). The reader should be aware of these two quite opposite
meanings and of what is implied by the psychologist when characterizing research as
"empirical".

3. A basic assumption of contemporary psychology is that the brain is the organ of the

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mind, a term which simply refers to the internal processes that determine the organization
of complex behaviour. Accordingly, one approach to the study of the properties of mind is
to study the functioning of the brain. Such research is sometimes correlational in that it
relates indices of brain function (e.g., electroencephalograms [EEG], evoked potentials)
in humans and animals to behavioural processes. Often, however, and for reasons related
to the control and sorting out of relationships among variables discussed under point 1),
the research involves an experimental study of the effects of manipulation of the brain on
behaviour. No one assumes that the brains of experimental animals are miniature human
brains; however, it is assumed that the basic principles of brain organization are common
across mammalian species and that the brain of the particular animal species may serve
as a model for certain aspects of human brain function.

4. Psychologists incorporate and manipulate motivational variables in their research for

three somewhat dissimilar reasons. The first is when the subject of study is the
motivational system in question, e.g., the control of feeding or drinking behaviour. In
such a context, it is obvious to all why an animal might be on a water or food- restricted
regimen. The second is when the motivational system in question is being used as a
model for other appetitive or aversive motivational systems, as discussed under point 2).
A third reason, and one that is often not well appreciated by those outside the discipline,
is that such manipulation represents an effective means of facilitating the controlled study
of phenomena only indirectly related to the motivational manipulation.

Behaviour that is oriented towards obtaining access to food and water, or escaping or
avoiding an aversive event, can form the basis of sound inferences about non-
motivational processes associated with, for example, learning, memory or perception.
The origin of the misunderstanding is that, although motivation is manipulated, the
dependent variables of the study relate to learning. The misunderstanding is compounded
by the fact that although sometimes the interest is in learning as a process itself, other
times it is in learning as a process that is affected by sensory, perceptual, motor or other
processes that are in fact the principal foci of the study. As a simple example, consider the
case of studying the capacity of an animal for pattern discrimination. One approach might
be to place the animal repeatedly into a chamber with two doors. Behind the door with
pattern A, food is always to be found. Behind the door with pattern B, there is no food.
The animal is then taught to enter one of the doors each time it is placed in the chamber.
To learn to enter the door behind which the food is located requires the animal to be
motivated and interested in finding and consuming the food, and that is why motivation is
manipulated. When the animal can consistently perform the correct response, i.e., when it
has learned to go to the door with pattern A, one is in a position to make some statement
about the perceptual capability of the animal, even though what was measured was
learning, and what was manipulated was motivation. The analysis of learning is central to
experimental psychology, and motivation is necessary for that analysis. The rationale for
the choice of motivation manipulated is not always obvious, but the later discussion of
Guideline 7 attempts to identify some of the considerations that are made. Often these
considerations relate to practicability and minimization of behavioural variability, which
promotes the economical and efficient use of animal resources.

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Over the years, members of the general public, the discipline of psychology itself, and
other disciplines, have expressed concerns about aspects of psychological research
involving animals. In part, this may be due to psychology including as subjects of study
phenomena to which one can easily relate on a highly personal level (e.g., stress, pain,
anxiety, motivation, etc.). In part, this may be due to a poor understanding of the nature
of the model systems as a research tool, and we have tried to address that in these
introductory remarks. However, in large measure, this is undoubtedly due to a language,
vocabulary, or jargon used to characterize phenomena, events, or hypothetical processes
that can sometimes conjure unrealistic, distressing images. Those with concerns are to be
encouraged to become fully informed about the nature of psychological research, and
those engaged in research are to be encouraged to be sensitive to these concerns and to
discuss openly their work and its implications with concerned individuals outside their
discipline.

Certain procedures used in the study of psychological problems undoubtedly do produce

some distress in animal and human subjects, and this places directly on the investigator a
responsibility to question seriously what is being done, why it is being done, and where
the work will lead. The Canadian Council on Animal Care (CCAC) and the scientific
community have had a continuing concern for these and other issues related to the care
and use of experimental animals. In the recognition that there are certain aspects of
behavioural research that are misunderstood and can be problematic with respect to
research ethics, the Canadian Psychological Association (CPA), in consultation with the
CCAC, has prepared the following set of guidelines to assist psychologists as researchers
and instructors in making the ethical decisions viewed to be an integral part of
behavioural research with animals. It is hoped that these guidelines will complement
those of the CCAC and facilitate the conduct of ethically responsible research that will
promote our understanding of basic processes underlying behaviour.

GUIDELINES FOR THE USE OF ANIMALS IN RESEARCH

AND INSTRUCTION IN PSYCHOLOGY:
COMMENTARY AND ELABORATION

The discipline and profession of psychology in Canada shares with contemporary society
a deep concern for the welfare and humane treatment of animals, especially in scientific
research and instruction. While it is recognized that animal research is essential to the
further development of scientific knowledge, it is also recognized that there are limits to
what should be done to animals in the conduct of that research. In response to this
concern, the Canadian Psychological Association (CPA), in consultation with the
Canadian Council on Animal Care (CCAC), has formulated these guidelines to assist
psychologists as researchers and instructors in making the ethical decisions that are an
integral aspect of working with animals. These have been published in a form suitable for
posting in laboratories. The purpose of this commentary is to provide some elaboration of
considerations that psychologists should give with respect to implementation of the
preamble and each guideline.

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Psychologists have an obligation to advance knowledge and promote welfare
through the competent conduct of research, the accurate communication of findings,
and the effective instruction of students. However, their values and goals as scientists
sometimes come into conflict with their values related to the treatment of living
organisms. Dilemmas posed by the conflict cannot be resolved by rigid rules and
regulations, but require a careful weighing of values and alternatives. In many
cases, the decisions reflect a relative judgement of the value of the research and the
effects of the procedures on the animals. Psychologists using animals for research or
instruction should be prepared to make such decisions and to explain the bases of
their decisions to an informed audience. The following guidelines are intended to
assist the scientist in making these ethical decisions.

The CPA's Guidelines for the Use of Animals in Research and Instruction in Psychology
were formulated from the ethical perspective advocated by Diener and Crandall (1978a)
in their book Ethics in Social and Behavioural Research. They consider research ethics to
be a process of decision-making rather than one of devising explicit rules and regulations
intended to govern the conduct of all research under all circumstances. Their approach is
well represented in the following passage:

"The ethical or moral scientist makes individual judgements about research practices in
light of his/her own values. According to this approach to ethics, the moral person is not
one who blindly follows ethical codes, no matter how enlightened. The ethical decision-
maker is one who realizes that his/her choices are related to values, and weighs these
values carefully when making important decisions. For the moral person there may be a
few moral absolutes (Szasz, 1967); however, he or she realizes that most moral decisions
must be made individually in each case (Smith, 1969). This meaning of ethics emphasizes
the process by which the decisions are made as well as the final choice. The decision is
made by a person who is educated about ethical guidelines, carefully examines moral
alternatives, exercises judgement in each situation and accepts responsibility for
his/her choices (Diener and Crandall, 1978b) (emphasis added)."

As reflected in the Preamble to these Guidelines, this ethical framework clearly places the
responsibility for the careful consideration of personal, social, and professional values on
the individual scientist. A corollary of this is that the individual researcher or instructor
must accept responsibility for choices through accountability to an informed public.
Specifically, the investigator must be able to explain the rationale for both the research
problem and its methodology to informed colleagues, peers, and institutional committees.
If necessary, the investigator must be able to defend the research against the criticisms
that suffering inflicted on animal subjects was unnecessary in view of the objectives of
the research, or unconscionable in view of the balance between the suffering inflicted and
expectation of gain in scientific knowledge or education. The present commentary is
intended to provide the scientist with an indication of problematic issues and concerns
that require special attention and consideration.

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A. THE SCIENTIST

1. Prior to undertaking a research or instructional project with animals, the

scientist has a responsibility to be sufficiently knowledgeable to ensure compliance
with these guidelines. When in doubt about compliance, the scientist should consult
with informed colleagues and the institutional Animal Care Committee (ACC) and
give due regard to their advice. Investigators are reminded that it is the policy of the
major granting agencies, e.g., Medical Research Council (MRC) and the Natural Sciences
and Engineering Research Council (NSERC), of government departments and of most
universities that ACC approval must be received prior to commencing any project with
animals.

The position that the ethical scientist is one who can make informed decisions and who is
prepared to give a reasoned judgment for the values and appropriateness of the objectives
and procedures of the research or teaching assumes a considerable depth and breadth of
knowledge by the scientist. The decisions involved are usually complex and multifaceted.
If they are to be informed, scientists must be familiar with the recent literature relevant to
the problem, aware of the current status of the problem, familiar with procedures
involved, either through study of the literature or direct experience with the techniques,
and aware of potential risks. When undertaking research in a new area, or when the
research involves severe stress, pain, or privation, the investigator may have doubts about
the breadth and depth of his/her knowledge and experience. Under such circumstances,
there is an obligation to consult with informed colleagues and/or the ACC and to give due
regard to their advice. Such consultation does not absolve the investigator from
responsibility for the decisions; however, it is evidence of a responsible attitude towards
becoming sufficiently knowledgeable to undertake research in an ethical manner.

2. A scientist trained in research methods and experienced in the care of

laboratory animals should ensure that the comfort, health and humane treatment of
experimental animals are given appropriate consideration.

Psychological research frequently requires that an investigator interact with animal

subjects over an extended period of time. Accordingly, the psychologist has a vested
interest, quite apart from humane considerations, in ensuring that experimental animals
are well treated and healthy; otherwise, it is likely that behavioural data will be unreliable
and the research objectives not achieved. Although the day-to-day maintenance and/or
behavioural testing of animals may be done by trained technical staff, it is the
responsibility of the individual scientist to be able to recognize good and poor practices
by staff and students. This ability requires training in research methods and experience in
animal care procedures. While our attention is naturally drawn to practices that involve
pain or physical illness, the scientist should be sensitive to problems that can arise from
the capacity of some animals to form human attachments (e.g., the distress that might be
experienced by the animal when returned to isolated quarters at the end of a prolonged
study).

3. The scientist should ensure that all individuals under his/her supervision have

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the training and competence needed to carry out their responsibilities for
experimental procedures, care, maintenance, and handling of the species being used.

A variety of technical skills is required in any laboratory to ensure proper care and
handling of animals, and it is the responsibility of the scientist to ensure that all
supervised individuals have the skills and attitudes required to carry out competently their
assigned duties. Since every species has unique biological and social needs, the design of
experimental and maintenance protocols should take into consideration the species'
normal ecology, evolutionary history, and behavioural adaptations to the natural
environment. It is the responsibility of the scientist to acquire sufficient expertise in these
regards, and to ensure that the training of technical staff, students, and research associates
is adequate to meet their respective responsibilities. This is necessary, not only from the
point of view of the humane treatment of animals, but also from the scientific perspective
of generating reliable data.

4. The scientist should be fully cognizant of the CCAC's guidelines and of current
federal, provincial, and local laws and regulations concerning the acquisition, care,
use, and disposal of animals.

This guideline is self-explanatory. As professionals and as members of society,

psychologists have a responsibility to be aware of and to follow federal, provincial, and
local laws and regulations concerning animals. In cases of doubt, the scientists should
consult with the chairman of the ACC or with the CCAC. Compliance with this Guide is
a requirement of Canada's major granting agencies, many journals in Canada, and ACCs.

B. RESEARCH

5. There must be a reasonable expectation that studies involving animals will: a)

increase understanding of structures and processes underlying behaviour; b)
increase understanding of the particular animal species used in the experiment; or
c) result eventually in benefits to the health and welfare of humans or other animals.

This guideline outlines the general spheres to which psychological research should
contribute if animals are legitimately to be involved. It specifically recognizes the value
of research, the implications of which are largely theoretical or philosophical, and is
consistent with the CCAC guidelines in that there is a reasonable expectation that the
development of new scientific knowledge and conceptualization may result in eventual
benefits to the health and welfare of humans or other animals.

This guideline refers more to research programs than to individual studies. It is intended
to recognize that there is no such thing as the "definitive study" and that the significance
of any individual experiment, especially when viewed after the fact, is not always
immediately apparent and can be easily trivialized. Accordingly, a particular experiment

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must be judged within the context of a research program as to whether it will contribute
in a meaningful way to a systematic empirical or theoretical base.

An integral part of any research program is the use of small pilot studies, the results of
which are at best suggestive, but which are important for decisions about directions to
proceed, research design, parameters, etc. The value of such studies is often indirect, and
therefore must be evaluated in the broader context of a research.

This guideline bears also on the problem of replication or reproducibility of results in

psychological research. Replication is a necessary and desirable aspect of science when it
is seen as a manipulation of a special class of factors (e.g., different laboratories, different
experimenters, different times of the year, etc.), which can provide new scientific
knowledge of value in understanding a phenomenon. As with the case of pilot studies,
replications must be seen within the broader context of a research program. To the extent
that there is a reasonable expectation that a replication will contribute to new scientific
knowledge, it is consistent with this guideline.

6. Procedures subjecting animals to pain, stress, privation, or death should be

used only when an acceptable alternative procedure is unavailable.

In psychological research, the subjection of animals to procedures involving pain, stress,

privation, or death occurs in two broad contexts. The first is when the subject of study is
pain, stress, motivational systems, or aspects of death, all of which are legitimate and
important areas of the discipline. In such a context, there are seldom realistic alternatives,
and the attention of the scientist must focus on ways of minimizing discomfort. For
example, the study of stress must necessarily involve manipulations that will produce
stress; nevertheless, the investigator must consider ways to minimize the trauma of these
manipulations. In the second context, procedures are employed to induce motivational
states that facilitate the controlled study of phenomena only indirectly related to the
motivational manipulations (for example, learning discrimination, memory, sensory
thresholds, etc.). Under such circumstances, the scientist has an ethical obligation to
consider whether the research objectives could be met using broad alternatives not
involving pain or discomfort.

7. Scientists should examine methodological and procedural techniques for the

purpose of minimizing discomfort, illness, and pain to animals.

When acceptable alternative procedures to ones that involve pain or privation are not
available, there remains a responsibility to examine methodologies and procedures that
will minimize discomfort, illness, or pain, and that are consistent with the objectives of
the research. The judgement involved clearly requires considerable knowledge of the
species and its behavioural repertoire, as well as the research problem. Issues that may
arise in considering this guideline include: Is the species appropriate for the study? Have
the motivational systems and the biological/social needs of the species been assessed so
that a reasonable judgment can be made about the relative discomfort and stress produced
by various potentially painful procedures or different kinds of levels of privation? Can

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motivational states in the given species be better controlled through appetitive motivation
(e.g., water or food deprivation) or through aversive motivation (e.g., mild electric
shock)? Have parameters of the aversive stimuli or the deprivation been selected
judiciously so as to be optimal in light of the behavioural requirements of the research
and the principle of minimizing discomfort? Could lower levels of shock or privation be
used and still produce reliable behaviour? Could a lower level of aversive stimulation or
privation be used, even though more animals might be required because of less reliable or
stable behaviour? In studies involving food motivation, is privation to a fixed percentage
(e.g., 80%) of ad lib body weight required, or would controlled daily access to food (e.g.,
every 23 hours), perhaps in combination with a preferred incentive, be sufficient?

Manipulations involving surgery can be problematic with respect to discomfort, and

issues that may arise in this regard include: Is the method of an anesthetization optimal?
Are the surgical procedures sufficiently aseptic to minimize post-operative infection and
other stress? Is an analgesic required during post-operative recovery? Does the
manipulation, such as implantation of a chronic cannula or electrode assembly, cause
irritation, and if so are there ways to minimize this? Does a physiological or
pharmacological manipulation (including administration of toxins) cause a generalized
deterioration of the well-being of the animal, and if so, how can this be minimized in a
manner consistent with the research objectives?

It is sometimes suggested that one approach to minimizing discomfort is to minimize the

number of animals used in the research. However, since the detection of treatment effects
is against a background of uncontrollable behavioural variance, one must be careful to
ensure that there is sufficient power to detect those effects when they are there.
Otherwise, the animals that were used would have suffered needlessly. However,
investigators should consider single-subject designs, repeated measures designs, and
other techniques to minimize variance, all of which could lead to fewer total subjects
being required for the research.

8. An experiment should be terminated whenever it becomes apparent to the

scientist or the institutional ACC, that its continuation will result in injury or
suffering that is incompatible with these guidelines.

With the best of planning, some experiments simply do not work out as expected. This
can be due to various reasons, such as equipment failure, procedures having unforeseen
effects, experimenter error, or behavioural variance so large as to obliterate any possible
treatment effects. Also, on occasion, new research results become known and may make
ongoing research redundant. In such situations, the experimenters must give careful
consideration to whether it is worthwhile to continue the work, and do so only if they are
satisfied that it is justified in light of these guidelines.

9. The killing or other disposition of experimental animals at the termination of

the experiment must be accomplished in a humane manner.

At the end of most experiments in psychology, experimental animals are killed. This must

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be done in a humane manner, as described in this Guide, and in Volume 2 (CCAC, 1984).
While this is obvious and is the accepted practice, problems can arise when research
animals are not killed. For example, releasing trapped animals back into the wild may or
may not be humane depending on the species, its territorial behaviour, feeding habits,
time of year, etc. As noted earlier, some animals can form human attachments and
problems can arise after experiments involving long-term interaction with them if they
are returned to isolated laboratory housing. The responsibility of the scientist to his/her
animals extends beyond the actual termination of the experiment, and careful
consideration must be given to whether the means of disposing of the experimental
animal is actually humane.

C. INSTRUCTION

10. The decision to use animals for instructional purposes must be based on a
consideration of educational objectives rather than contributions to new scientific
knowledge. In other respects, ethical practices in the care and treatment of animals
are the same as those that apply to the use of animals in research.

A committee of the British Psychological Society (BPS) has commented that, "...No
psychology undergraduate can for long remain unaware of the extent to which the
empirical basis of much psychological theory is derived from experimental work with
animals. Accordingly, it is appropriate that, as a matter of course, all undergraduate
students of psychology should receive specific instruction on the issues which arise from
animal experimentation, issues scientific, intellectual, methodological, practical, and
ethical" (BPS, 1979).

The CPA shares the view that instruction on the use of animals in psychology is desirable
and necessary. On occasion, the actual use of animals in instruction is required to achieve
educational objectives. In such cases, the same general considerations must apply to
animal use in instruction as in research, that is, a balancing of the expected benefits
against the costs, with the benefits seen in terms of advancing education rather than
incrementing scientific knowledge. Clearly, classroom demonstrations of animal
behaviour by their very nature involve phenomena which may not intrinsically advance
scientific knowledge. However, to the extent that they assist the student's understanding
of existing knowledge, they make a substantive contribution. As contained in the BPS
position, there is an obligation to incorporate material on ethical issues into discussions of
animal use. In this way, instructors can, by example, promote the ethical use of animals
by future scientists and instill in students an appropriate sensitivity to associated issues.

11. Classroom demonstrations involving animals should only be used when

instructional objectives cannot be achieved through the use of videotapes, films, or
other alternative methods. Careful consideration should be given to whether the
type of demonstration is warranted by the anticipated instructional gain.

Videotapes and films represent effective means of demonstrating principles of animal

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behaviour and experimentation. However, there are often advantages of using real
animals, not the least of which is to convey the realism of the phenomenon. In deciding
on the medium of instruction, instructors have an obligation to evaluate carefully their
instructional objectives and

In making the evaluation, the instructor should be sensitive to the possible trauma that the
animals may experience in being brought into a classroom, and to the possibility of
disease transmission to or from the animal. The instructor must also consider whether the
animal will be used solely for the demonstration, or whether it has already been used in
experimentation, or is breeding stock, or is maintained for the purpose of demonstrations.

The guideline makes reference to "type of demonstration" to alert the instructor to the
possibly adverse reactions that a demonstration, live or filmed, may produce in
unprepared students. Procedures which to the naive viewer may appear to involve pain or
stress (e.g., showing animals with chronic cannulae or electrode assemblies, animals
having epileptic seizures, animals being operated on, injected with drugs or social
animals raised in isolation, etc.) are especially problematic.

12. Student projects involving pain or distress to animals should be undertaken

judiciously and only when the training objectives cannot be achieved in any other
way.

Student research projects fall along a continuum from casual classroom projects at one
end, to doctoral dissertation research (which should contribute to new scientific
knowledge) at the other. The value of such projects is seldom to be found in their making
substantive new contributions to knowledge, but is found in their advancing knowledge
through communication. If students are to acquire the knowledge and expertise required
by these guidelines, it is necessary that they gain experience in working with animals.
Especially in the case of students who show every indication of embarking on a research-
oriented career, that experience may involve using procedures that involve minimal pain
or distress. If so, the instructor must consider very carefully the appropriateness of the
project and its procedures against its training objectives for the individual student and the
development of student sensibilities towards animals. Students and instructors are
reminded that ACC approval must be received prior to commencing any project with
animals.

These guidelines shall be conspicuously posted in every laboratory teaching facility

and applied setting where animals are being used.

The CPA recommends that a copy of the guidelines and this commentary be included in
laboratory manuals as well as that selected sections be posted in all laboratory facilities.
It is hoped this will not be seen as an empty formality, but rather as an invitation for all

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laboratory personnel to become acquainted with the ethical process. Like science itself,
ethical procedures are advanced by communication and discussion.

In any situation involving decisions and judgment, there will, on occasion, be

disagreements and misunderstandings. When these arise in the context of the use of
animals in research or instruction in psychology, they should be resolved as quickly as
possible so as not to impede legitimate research nor to prolong unacceptable procedures.
The well-being of the animal must be a paramount concern. In general, there are two
classes of problems. In the first, there may be allegations by students, colleagues, or the
public that some on-going or completed research or instruction is inappropriate in light of
these guidelines. In this case, the concerned individual should attempt to work through
the ACC of the institution in which the research is carried out. In the second, individual
psychologists may find their ACC or a granting agency unwilling to approve some
research which they feel is scientifically and ethically warranted. The CCAC has
developed a formal appeal mechanism of which researchers may take advantage. Part of
this mechanism will involve consultation with the CPA.

D. REFERENCES

ABRAMSON, L.Y. and SELIGMAN, M.E.P. Modelling psychopathology in the

laboratory: History and rationale. In: Maser, J.D. and Seligman, M.E.P., eds.
Psychopathology: experimental models. San Francisco, CA: W.H. Freeman & Co., 1977:
1-26.

BRITISH PSYCHOLOGICAL SOCIETY. Policy statement. Brit. Psych. Soc. Bull. 1979;
32: 50.

CANADIAN COUNCIL ON ANIMAL CARE. Guide to the Care and Use of

Experimental Animals, Vol. 2, Ottawa, Ont.: CCAC, 1984.

DIENER, E. and CRANDALL, R. Ethics in social and behavioural research. Chicago, IL:
University of Chicago Press, 1978a.

DIENER, E. and CRANDALL, R. Ethics in social and behavioural research. Chicago, IL:
University of Chicago Press, 1978b: 4-5.

HEBB, D.O. Textbook on psychology. 2nd Ed. Philadelphia, PA: W.B. Saunders, 1966:
19.

SMITH, M.B. Social psychology and human values. Chicago, IL: Aldine, 1969.

SZASZ, T.S. Moral man: A model of man for humanistic psychology. In: Bugenthal, J.T.,
ed. Challenges of humanistic psychology. New York, NY: McGraw-Hill, 1967.

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 Chapter XIV - Guidelines for the use of Animals in Neuroscience Research

XIV. GUIDELINES FOR THE USE OF ANIMALS IN

NEUROSCIENCE RESEARCH*

A. INTRODUCTION

Research in the neurosciences contributes to the quality of life by expanding knowledge

about living organisms. This improvement in quality of life stems in part from progress
toward ameliorating human disease and disability, in part from advances in animal
welfare and veterinary medicine, and in part from the steady increase in knowledge of the
abilities and potentialities of human and animal life. Continued progress in many areas of
biomedical research requires the use of living animals in order to investigate complex
systems and functions because, in such cases, no adequate alternatives exist. Progress in
both basic and clinical research in such areas cannot continue without the use of living
animals as experimental subjects. The use of living animals in properly designed
scientific research is therefore both ethical and appropriate. Nevertheless, our concern for
the humane treatment of animals dictates that we weigh carefully the benefits to human
knowledge and welfare whenever animal research is undertaken. The investigator using
research animals assumes responsibility for proper experimental design, including ethical
as well as scientific aspects.

The scientific community shares the concern of society at large that the use of animals in
research should conform to standards that are consonant with those applied to other uses
of animals by humans. While it is unlikely that any particular set of standards will satisfy
everyone, it is appropriate for scientific societies to formulate guidelines that apply to the
humane use of laboratory animals in particular areas of research. Ideally, such guidelines
should also be acceptable to society at large as reasonable and prudent.

Most of the more specific sections of this document were formulated with respect to
research using warm-blooded vertebrates. As a general principle, however, ethical issues
involved in the use of any species, whether vertebrate or invertebrate, are best considered
in relation to the complexity of that species's nervous system and its apparent awareness
of the environment, rather than physical appearance or evolutionary proximity to humans.

B. FACTORS THAT RELATE TO THE DESIGN OF EXPERIMENTS

The primary factor used to evaluate humane treatment in animal research is degree of
distress or discomfort, assessed by anthropomorphic judgements made by reasonable and
prudent human observers. The fundamental principle of ethical animal research is that
experimental animals must not be subjected to avoidable distress or discomfort. This
principle must be observed when designing any experiment that uses live animals.

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Although most animal research involves minimal distress or discomfort, certain valid
scientific questions may require experimental designs that inevitably produce these
effects. Such situations, while uncommon, are extremely diverse and must be evaluated
individually. It is critical that distress and discomfort be minimized by careful
experimental design. It is also important to recognize that there is no difference between
distress and discomfort that may be inherent in a valid experimental design and that
which may occur as an unintended side effect. It is therefore incumbent on the
investigator to recognize and to eliminate all avoidable sources of distress and discomfort
in animal subjects. This goal often requires attention to specifics of animal husbandry as
well as to experimental design.

Invasive procedures and paralytic drugs should never be employed without benefit of
anesthetic agents unless there is a very strong scientific justification and careful
consideration is given to possible alternatives. Advances in experimental techniques, such
as the use of devices chronically implanted under anesthesia, can offer alternative
approaches. If these are not feasible, it is essential to monitor nociceptive responses (for
example, recordings of EEG, blood pressure and pupillary responses) that may indicate
stress in the animal subject, and to use these as signals of the need to alleviate pain, to
modify the experimental design, or to terminate the experiment.

When designing research projects, investigators should carefully consider the species and
numbers of animals necessary to provide valid information, as well as the question of
whether living subjects are required to answer the scientific question. As a general rule,
experiments should be designed so as to minimize the number of animals used and to
avoid the depletion of endangered species. Advances in experimental methods, more
efficient use of animals within-subject designs, and modern statistical techniques all
provide possible ways to minimize the numbers of animals used in research. This goal is
completely consistent with the critical importance of replication and validation of results
to true progress in science.

C. FACTORS THAT RELATE TO THE CONDUCT OF EXPERIMENTS

Research animals must be acquired and cared for in accordance with the guidelines
published in the NIH Guide for the Care and Use of Laboratory Animals (National
Institutes of Health, Publication No. 85-23, Revised 1985). The use of an animal
scheduled for euthanasia by a pound or shelter saves the life of another; therefore, the use
of pound or shelter animals is endorsed for research projects in which they are suitable
subjects. In using animals acquired from a pound or shelter, as with all other aspects of
research, investigators must adhere to the relevant local, state and federal law. (The
reference to pound or shelter animals was added to the Guidelines following the
recommendation by the Committee on Animals in Research and approval by Council.)
The quality of research data depends in no small measure on the health and general
condition of the animals used, as well as on the specifics of the experimental design.
Thus, proper animal husbandry is integral to the success of any research effort using
living animal subjects. General standards for animal husbandry (housing, food quality,

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ventilation, etc.) are detailed in the NIH Guide. The experienced investigator can
contribute additional specifics for optimum care for particular experimental situations, or
for species not commonly encountered in laboratory settings.

Surgery performed with the intent that the animal will survive, (for example, on animals
intended for chronic study), should be carried out, or directly supervised, by persons with
appropriate levels of experience and training, and with attention to asepsis and prevention
of infection. Major surgical procedures should be done using an appropriate method of
anesthesia to render the animal insensitive to pain. Muscle relaxants and paralytics have
no anesthetic action and should not be used alone for surgical restraint. Post-operative
care must include attention to minimize discomfort and the risk of infection.

Many experimental designs call for surgical preparation under anesthetic agents with no
intent that the animal should survive. In such cases, the animals ordinarily should be
maintained unconscious for the duration of the experiment. At the conclusion of the
experiment, the animal should be killed without regaining consciousness and death
ensured before final disposition.

Certain experiments may require physical restraint, and/or withholding of food or water,
as methodological procedures rather than experimental paradigms. In such cases, careful
attention must be paid to minimize discomfort or distress and to ensure that general
health is maintained. Immobilization or restraint to which the animals cannot be readily
adapted should not be imposed when alternative procedures are practical. Reasonable
periods of rest and readjustment should be included in the experimental schedule unless
these would be absolutely inconsistent with valid scientific objectives.

When distress and discomfort are unavoidable attributes of a valid experimental design, it
is mandatory to conduct such experiments so as to minimize these effects, to minimize
the duration of the procedure and to minimize the numbers of animals used, consistent
with the scientific objectives of the study.

* Reproduced with permission of the Society for Neuroscience from Handbook for the
Use of Animals in Neuroscience Research (1992). References to American publications
and to American states are not relevant to Canadian researchers.

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 APPENDIX I

HOUSING AND ENVIRONMENT

The following charts are based on successful experiences and professional judgement
ever cognizant of the performance evaluation of the animals being maintained. As there
are few objective data available on the ideal space requirements for individual laboratory
species, veterinary judgement must be considered in evaluating housing requirements.
The Social and Behavioural Requirements of Experimental Animals chapter provides
additional guidelines for specific species.

SPECIES SPACE PER ANIMAL TEMPERATURE oC R.H. Ventilation B.T.U.

(weight) Single- Minimal Group Room/Cage Pen/Free Animal/
Floor or % Changes/ Hour
Area Height Loose o
C Ranging Hour
Housing
CAT >4 kg 0.28 m2 0.76 m 0.56 m2 20-22 15-25 45- 10-18 25-30
0.37 m2 Perch Perches 60
CATTLE
Calf 1.5 m2 2.4 m2 10-25 2-27 40- 5-20 300-800
Cow 3.0 m2 7-10 m2 70
CHICKEN Chapter IV Table 3 18-22 12-27 45- 5-15 30
70
DOG
<12 kg 0.75 m2 0.8 m 1.5 m2 18-21 5-25 45- 8-12 80-150
12-30 kg 0.9 m 2.0 m2 55
>30 kg 1.20 m2 pen-2.0 3.0 m2
m
2.23 m2
GERBIL 116 15 cm pair + 15-24 40- 8-10 4.0
cm2 litter 50
900 cm2
GOAT 1.4 m2 2.0 m 1.0 m2 15-24 7-30 55- 6-12 350-500
65
GUINEA
PIG 300cm2 18 cm 500 cm2 18-22 50- 4-8 5-6
<350 g 22 cm 800 cm2 60
>350 g 650cm2
HAMSTER fem. +
>100 g 100cm2 18 cm litter 21-24 45- 6-10 2.5

300
 18 cm 900 cm2 65
2
120cm
HORSE 4-5 cm2 3 m 13-17 m2 10-24 2-27 25- 4-8
75
MOUSE fem. +
2
<20 g 65 cm 13 cm litter 22-25 50- 8-12 0.6
>20 g 100cm2 15 cm 160 cm2 70
NON-
HUMAN
PRIMATE
Baboon
(Papio sp) 0.74 m2 0.91 m 2.8 m2 21-26 15-30 45- 12-16 60-140
5-12 kg 1.22 m 60
>12 kg 1.39 m2
(Macaca
sp) 0.4 m2 0.81 m 2-3 m2 22-25 18-29 45- 10-15 60-200
<7 kg 0.6 m2 0.91 m perches 60
7-15 kg 0.75 m2 1.2 m
>15 kg
OPOSSUM 0.56 m2 0.75 m 21-25 10-27 45- 10-12
65
PIGEON 0.18 m2 0.38 m 16-20 5-27 45- 12-15 1.2
70
QUAIL 400cm2 15 cm 200 cm2 21-22 20-30 45- 10-15
max. 30 70
cm
RABBIT fem. +
<4 kg 0.37 m2 0.40 m litter 16-22 10-28 40- 10-20 30-40
>4 kg 0.45 m 0.93 m2 shade 50
0.46 m2
RAT fem. +
<150 g 150cm2 18 cm litter 20-25 50- 10-20 4.0
>150 g 18 cm 800 cm2 55
250cm2
SHEEP Chapter IV Table 2 5-21 0-24 50- 15-25 500-800
75
SWINE 2.0 to 4.0 m2 of fem. + 17-24 10-25 55- 15-20 250-500
pen litter; 75
space per animal 5-8.5 m2

301
 APPENDIX II

BREEDING AND REPRODUCTION DATA

SPECIES Breedin Cycle Duration Breeding Gestatio Litter Optimal*** Light

g Age Type* of Behaviour n Size Hour
(age and range Length Sexual ** mean (d) and Reproducti s
weight) female- Receptivi and Range ve
male (days) ty Season** range Span
(yrs/mos)
CAT 7-10 14-24 3-8 d Ph (f. to 62 2-6 6-7 yrs 12-14
mos S.P. irregular m.) 57-65
Jan. to
Sept.
CATTLE 16-24 18-24 10-24 hrs Ph (A1) 283 1 8-10 yrs 8-12
mos P all year 279-290
CHICKE 4-6 mos Poly all 21 9-12 mos 13-14
N year incubatio
n
DOG 10-14 21 4-8 d Ph (f. to 63 breed 6-7 yrs 10-12
mos M m.) 58-68 4-10
Biannual depende
nt
GERBIL 9-12 4-6 14-18 hrs MP 25 4-5 15 mos 12-14
wks P all year 24-26
GOAT 15-18 14-21 48-72 hrs Poly 151 1-2 4-5 yrs 8-12
mos S.P. Sept. to 149-153
Feb.
GUINEA 3 mos 15-19 6-14 d H (1 to 6) 65 2-6 2 yrs 12-15
PIG P all year 59-72
HAMSTE 6-8 wks 4 6-20 hrs M.P. or 16 5-8 15 mos 12
R P H (1 to 5) 15-18
all year
HORSE 36-60 19-24 3 d Ph (f. to 335 1 12-15 yrs 8-12
mos S.P. 2-6 m.) 320-360
Feb. to
Aug.
MOUSE 6 wks 4-5 10-20 hrs H (1 to 4) 20 6-12 7-8 mos 14
P all year 19-21
NON- 48-66 31-32 3-4 day H (1 to 6) 175 1 5-20 yrs 12-14
HUMAN mos menses all year 154-185
PRIMATE day 15-17

302
 optimal
Baboon breeding
(Papio sp) period
(Macaca 36-48 28 3-4 day H (1 to 8) 165 1 5-20 yrs 10-14
sp) mos menses or Ph (f. to 150-180
day 10-12 m.) all year
optimal
breeding
time
OPOSSU 8-12 22-27 7-14 d M.P. marsupia 1-12 5 yrs
M mos P post- l
partum 12-13
PIGEON 6 mos M.P. 13 2 yrs 12-14
all year incubatio
n
QUAIL 6 wks H (1 to 3) 16 5-6 mos 14
or poly incubatio
all year n
RABBIT 6-9 mos induce ovulation Ph (f. to 31 6-10 3 yrs 12-14
d variable m.) 28-34
P all year
RAT 10-12 4-5 10-20 hr H (1 to 6) 21 7-14 9-10 mos 12-14
wks P all year 20-22
SHEEP 18-24 m 16-17 1-1 1/2 d Poly 145 1-3 4-5 yrs 12
S.P. mid-Sept. 144-148
to mid-Jan.
SWINE 9-11 21 2-3 d Poly 114 6-16 3-4 yrs 10-12
mos P all year 112-116

APPENDIX III

PHYSIOLOGICAL AND NUTRITIONAL PARAMETERS*

SPECIES Rectal Resp. Heart Average Urine Daily Feed Digestible

Rate/ Rate/ Daily Excreted Recommendatio
Temp. Mean Mean Water ns Protein**
and and Consumptio Daily
o
C (range (range n %
+0.5 ) )
CAT 38.5 31 150 150 ml 50-120 110-225 g 30
(20-40) (110- 100-200 ml

303
 226)
CATTLE 38.5 29 58 45-65 L 14-23 L. 7.5-12.5 kg 8.5-10
(26-35) (46-55)
CHICKEN 39.5 (12-36) 300 ad lib 85-115 g 13-17
(150-
400)
DOG 39.0 24 110 25-35 ml/kg 65-400 250-1200 g 20
(20-34) (77- body wt ml breed
138) breed dependent
dependen
t
FERRET 38.5 34 240 75-100 ml 26-28 ml 140-190 g 9.5
(33-36) (200-
400)
GERBIL 38.5 90 360 3-4 ml or few drops 10-15 g 15
(70- (260- green feed
120) 600)
GOAT 39.0 19 90 1.5-4 L 1-2 L 1-4 kg 15
(12-35) (70-
135)
GUINEA 39.0 86 280 12-15 ml/ 15-75 ml 20-35 g 25-30
PIG (42- (230- 100 g body + Vit. C supp.
104) 380) wt
HAMSTER 39.0 77 332 8-12 ml 6-12 ml 7-15 g 16
(35- (250-
135) 500)
HORSE 38.0 12 44 25-55 L 3-15 L 8-16 kg 5.5-14
(10-14) (23-70)
MOUSE 37.5 138 470 3-7 ml 1-3 ml 3-6 g 12
(94- (325-
163) 780)
NON-
HUMAN
PRIMATE 39.0 25 115 150-400 1-1.5 kg
Baboon (22-35) (105- 400-600 ml ml + Vit. C supp. 17
(Papio sp) 150)
Cynomolgus 39.0 40 220 350-950 ml 150-550 350-550 g 17
(30-54) (165- ml + Vit. C supp.
(M. 243)
fascicularis)
OPOSSUM 34.5 36-65 (140- 100-200 ml 85-150 g 20-25
220)

304
PIGEON 41.0 25-30 (140- 40-50 ml 25-75 g 10-15
244)
RABBIT 40 260 80-100 50-90
39.0 (32-60) (130- ml/kg ml/kg 75-100 g 14
325) body wt body wt
RAT 92 350
37.0 (70- (250- 20-45 ml 10-15 ml 10-20 g 12
115) 450)
SHEEP 25 76 400-1200
39.5 (20-34) (70-80) 600-1800 ml ml 1-2 kg 5
SWINE 40 70
39.0 (32-58) (60-75) 4.5-6.5 L 2.5-4.5 L 1.5-3 kg 14

* Averages and ranges derived from literature mean values for young adult animals
under various conditions (from various sources).

** Refers to (ideal or digestible protein required; crude protein (CP)) levels in most
prepared laboratory animal diets may be considerably higher.

References

FOX, J. Biology and diseases of the ferret. Lea and Febiger, 1988.

FOX, J., COHEN, B. and LOEW, F. Laboratory animal medicine. Academic Press, 1984.

HARKNESS, J. and WAGNER, J. The biology and medicine of rabbits and rodents. Lea
and Febiger, 1983.

HECKER, J.F. The sheep as an experimental animal. Academic Press, 1983.

MERCK VETERINARY MANUAL. 6th Ed. Merck and Co., 1986.

SWENSON, M. Dukes' Physiology of domestic animals. 10th Ed. Cornell Un. Press,
1984.

APPENDIX IV

HEMATOLOGY*

SPECIES RBC Hb PCV Platelets WBC Neutrophils Lymphocytes Blood

X g/L Vol.
1012/L L/L X 109/L X 109/L 109/L (ml/kg)

305
 109/L
CAT 7.5 120 0.37 12.5 7.5 4.0 66.7
5.0- 80- 0.24- 190-400 5.5- 2.5-12.5 1.5-7.0 45-75
10.0 150 0.45 19.5
CATTLE 7.0 110 0.35 8.0 2.0 4.5 57
5.0- 80- 0.24- 220-640 4.0- 0.6-4.0 2.5-7.5 55-60
10.0 150 0.46 12.0
CHICKEN 3.0 90 0.3 12.0 3.0-6.0 14.0 83
2.5- 70- 0.22- 14-60 12.0- 7.0-17.5 60-90
3.5 130 0.35 30.0
DOG 6.8 150 0.45 11.5 7.0 2.8 83-101
5.5- 132- 0.38- 145-440 6.0- 3.9-12.0 1.0-4.8
8.5 193 0.57 17.0
GERBIL 8.5 150 0.48 4.3- 0.3-4.1 3.2-9.7 60-85
7.0- 121- 0.41- 638 21.6
10.0 169 0.52
GOAT 13.0 100 0.35 9.0 3.2 5.0 70
8.0- 80- 0.24- 250-750 4.0- 1.2-7.2 2.0-9.0 55-80
18.0 120 0.48 13.0
GUINEA PIG 5.2 110- 0.43 3.8- 2.6 6.4-7.5 65-90
4.8- 140 0.37- 450-630 13.5 2.0-3.1
5.9 0.46
HAMSTER 7.5 168 0.5 7.6 1.5-3.5 6.1-7.0 65-80
5.0- 146- 0.46- 300-570 5.0-
9.2 200 0.52 10.0
HORSE 9.0 144 0.41 9.0 4.7 3.9 72
6.8- 111- 0.32- 80-397 5.4- 2.3-8.6 1.7-6.8 75-100
12.9 190 0.53 14.3
MOUSE 9.1 110- 0.37- 5.0- 0.4-2.7 7.1-9.5 70-80
7.9- 145 0.46 600- 13.7
10.1 1200
NON-HUMAN
PRIMATES
Baboon (Papio 5.0 120 0.42
sp) 4.0- 90- 0.36- 135-400 3.0- 2.7-7.3 2.6-5.9 50-70
6.0 150 0.49 11.4
CYNOMOLGUS
5.0
(M. fascicularis) 3.9- 116- 0.38- 90-140 8.1- 1.3-8.1 3.5-8.3 55-75
7.1 145 0.50 21.3
OPOSSUM 5.0 0.42
3.4- 121- 0.30- 235- 3.0- 1.5-6.5 1.9-9.2 45-65

306
 7.1 198 0.58 1235 27.0
QUAIL 4.7 0.42
4.0- 110- 0.3- 12.5- 2.5-5.0 5.0-7.0 55-75
5.5 150 0.45 25.0
RABBIT 6.5 0.40 468
4.5- 94- 0.31- 180-750 4.0- 3.0-5.2 2.8-9.0 57-65
8.5 175 0.50 13.0
RAT 5.4- 115- 0.37- 450-885 4.0- 1.3-3.6 5.6-8.3 50-65
8.5 160 0.49 10.2
SHEEP 12.0 115 0.35 8.0 2.4 5.0
9.0- 90- 0.27- 250-750 4.0- 0.7-6.0 2.0-9.0 58-66.4
15.0 150 0.45 12.0
SWINE 6.5 130 0.42 16.0
5.0- 100- 0.32- 300-700 11.0- 4.0-7.5 6.0-10.0 52-69
8.0 160 0.50 22.0

* The normal values may vary according to age, sex, breed and function of animals.

References

ARCHER, R.K., JEFFCOTT, L.B. and LEHMANN, H. Comparative clinical

haematology. Blackwell Scientific Publications, 1977.

FOX, J.G., COHEN, B.J. and LOEW, F.M. Laboratory animal medicine. Academic Press,
1984.

HAWKEY, C.M. Comparative mammalian haematology; cellular components and blood

coagulation of captive wild animals. William Heinemann Medical Books Ltd., 1975.

JAIN, N.C. Schalm's Veterinary Haematology. 4th Ed. Lea and Febiger, 1986.

MITRUKA, B.M. and RAWNSLEY, H.M. Clinical biochemical and hematological

reference values in normal experimental animals. New York, Paris, Barcelona, Milan:
Masson Publishing USA Inc., 1977.

SWENSON, M.J. Dukes' physiology of domestic animals. 10th Ed. Cornwell Un. Press,
1984.

UNIVERSITY OF GUELPH. Veterinary teaching hospital (Manual), 1988.

307
 APPENDIX V

CLINICAL BIOCHEMISTRY REFERENCE VALUESa

Protein Asparta Alanine Alkaline

SPECIES Glucose Urea Cholest Tota Albu Globu te Phospha
mmol/L erol l min lin Amino- Amino- tase
mmol/L Total g/L g/L g/L transfer transfer U/L
mmol/L ase ase
(AST, (ALT,
SGOT) SGPT)
U/L U/L
CATb 3.89- 14.28- 2.46- 54- 21-33 26-51 26-43 6-83 25-93
6.11 21.42 3.37 78 (272) (397) (359) (2616) (5035)
(5.050. (66
42) 7)
CHICKENb (9.30) (56) (25) (31) (175)
COWb 2.5-4.16 14.28- 2.07- 67- 30-35 30-35 78-132 14-38 0-488
21.42 3.11 75 (331) (322) (10527 (2714) (194126
(3.190. (71 ) )
38) 2)
DOGb 3.61- 7.14- 3.50- 54- 26-33 27-44 23-66 21-102 20-156
6.55 19.99 6.99 71 (292) (345) (3312) (4726) (6636)
(5.050. (12.142 (4.610. (61
67) .86) 98) 5)
GOATb 2.78- 7.14- 2.07- 64- 27-39 27-41 167-513 24-83 93-387
4.16 14.28 3.37 70 (333) (365) (21976)
(3.490. (10.711 (69
39) .43) 5)
GUINEA PIGc 4.94- 15.35- 48- 24-27 46-48 38-45 66-74
5.29 17.99 56 (25) (47) (41) (70)
Hartley (500- (5.12) (16.67) (52)
800g)
HAMSTERc 3.61- 14.85- 4.71- 64- 32-37 53-124 21-50 8-18

308
Syrian (100g) 4.07 21.49 6.13 73 (352) (7932) (3511) (135)
(3.84) (18.333. (5.42) (675
08) )
HORSEb 4.16- 7.14- 1.94- 52- 26-37 26-40 226-366 3-23 143-395
6.39 17.14 3.89 79 (313) (337) (1411) (244101
(5.300. (2.880. (63 (29670 )
47) 04) 6) )
MOUSEd 9.71- 12.14- 1.27- 42- 21-34 18-82 55-251 28-184 28-94
CD-1 18.60 20.59 2.48 60 (28) (22) (139) (95) (67)
[Crl:CD- (15.00) (16.07) (1.89) (51)
1(ICR)BR]e
CF-1 [Crl:CF- 9.10- 8.57- 2.72- 54- 30-40 18-31 30-314 76-208 67-303
1BR]e 20.48 19.99 4.16 65 (35) (24) (177) (143) (167)
(14.46) (14.99) (3.49) (60)
B6C3F1 7.6-26.0 4.3-13.5 1.53- 47- 26-34 17-29 0-111 46-289
[B6C3F1/CrlB (7.85) 3.63 60 (30) (22) (43) (207)
R]f (17.3) (2.29) (52)
NON- (6.721. (63 (374) (253) (164)
HUMAN 16) 6)
PRIMATE
Baboon
(Papio sp)c
Cynomolgus 2.20- 3.80- 1.91- 68- 34-45 27-47 9-68 0-138 102-1163
(M. 4.70 10.00 4.52 86
fascicularis)g
Rhesus (3.890. 12.07- 3.31- 66- 43-44 27-79 27-42
c
(M. mulatta) 57) 14.85 4.43 80 (5527) (35)
(13.46) (3.87) (70 (149)
8)
PIGb 4.72- 7.41- 0.93- 79- 53-64 32-84 31-58 118-395
8.33 21.42 1.40 89 (596) (6126) (4514) (19484)
(6.610. (84 (267)
96) 7)
RABBITb 2.78- 0.14-
5.18 1.86
(4.080. (10.212. (0.690. (64 (273) (47) (79) (12014)
53) 14) 41) 3)
RATd 4.71- 11.42- 1.20- 63- 33-49 24-39 39-92 17-50 39-216
Wistar[Crl: 7.33 19.28 2.38f 86 (47) (31) (64) (32) (123)
(W)BR]h (6.22) (14.64) (1.79) (73)
F-344i 4.24- 7.85- 0.54- 60- 34-43 24-35 56-436 108-375 147-399
[CDF(F- 20.04 19.99 2.22 78 (39) (29) (233) (248)

309
344)CrlBR] (10.85) (10.00) (1.29) (66) (232)
CD[Crl:CD(S 5.55- 9.28- 1.18 59- 28-44 26-39 39-262 110-274 46-264
D)BR]j 16.71 22.13 (0.52- 79 (38) (32) (129) (161)
(11.69) (14.64) 1.914) (70) (216)
SHEEPb 2.78- 5.71- 1.34- 60- 24-30 35-57 68-387
4.44 14.28 1.97 79 (272) (445) (178102
(3.800. (1.660. (72 (30743 (304) )
33) 31) 5) )

a
Ranges with the means and standard deviations in parenthesis. Reported in S.I. units.
b
KANEKO, J.J., ed. Clinical chemistry of domestic animals. Academic Press, 1989: 886-
891.
c
LOEB, W.F. and QUIMBY, F.W., eds. The Clinical Chemistry of Laboratory Animals.
Pergamon Press, 1989: 417-476.
d
Sexes combined, 19-21 weeks.
e
Baseline haematology and clinical chemistry values for Charles River outbred mice:
Crl:CD-1(ICR)BR. Crl:CF-1BR. Charles River Laboratories Techn. Bull., 1986.
f
Values from Parke Davis Research Institute, Mississauga, Ontario.
g
CLARKE, D., TUPASI, G., WALKER, R. and SMITH, G. Stability of serum
biochemical parameters in Beagle Dogs and Cynomolgus monkeys. Clin. Chem. Newsl.
(In press).
h
Baseline haematology and clinical chemistry values for Charles River Wistar rats (CRL:
(W)BR) as a function of sex and age. Charles River Techn. Bull., Vol. 1, No. 2, 1982.
i
Baseline haematology and clinical chemistry calues for Charles River Fischer-344 rats -
CDF(F-344)CrlBR as a function of sex and age. Charles River Techn. Bull., Vol. 3, No. 1,
1984.
j
Baseline haematology and clinical chemistry values for Charles River
CD[Crl:CD(SD)BR] rats as a function of sex and age. Charles River Techn. Bull., Vol. 3,
No. 2, 1984.

310
 APPENDIX VI

SERUM ELECTROLYTE REFERENCE VALUESa

SPECIES Sodium Potassiu Chlorid Bicarbon Phosphor Calcium Magnesiu

mmol/L m e ate us mmol/L m
mmol/L mmol/L mmol/L mmol/L mmol/L
CATb 147-156 4.0-4.5 117-123 17-21 1.45-2.62 1.55-2.55
(152) (4.3) (2.00)
(2.06+0.2 (0.90)
4)
CHICKENb (2.52) (7.10)
COWb 132-152 3.9-5.8 97-111 17-29 1.81-2.10 2.43-3.10 0.74-0.95
(142) (4.8) (104) (0.84+0.1
(2.78+0.1 0)
5)
DOGb 141-152 4.37- 105-115 18-24 0.84-2.00 2.25-2.83 0.74-0.99
5.35 (1.39+0.2 (0.86+0.1
9) (2.55+0.1 2)
5)
GOATb 142-155 3.5-5.4 99-110 2.88-3.20 0.90-0.31
(150+3.1 (105+2. (1.03+0.1
) 9) (4.62+0.2 (2.58+0.1 2)
5) 5)
GUINEA PIGc 122-125 4.7-5.3 92-97 22-24 1.71-1.72 2.40-2.67 0.97-1.01
Hartley (500-
800g)
HAMSTERc 128-145 4.9-5.1 94-99 1.71-2.13 2.60-3.09 0.91-1.03
Syrian (100g)
(30+2.9)
b
HORSE 132-146 2.4-4.7 99-109 20-28 1.00-1.81 2.80-3.40 0.19-1.15
(139+3.5 (3.5+0.6 (104+2. (1.03+0.1
) ) 6) (3.10+0.1 3)
4)
MOUSEd
CD-1
[Crl:CD- 143-150 3.8-10.0 96-111 2.68-3.62 2.77-3.02

311
1(ICR)BR]e (148) (6.3) (105) (3.08)
(2.90)
CF-1 [Crl:CF- 139-157 4.8-8.9 104-119 2.91-4.65 2.25-2.89
1BR]e (148) (6.9) (111) (3.76)
(2.57)
NON-HUMAN
PRIMATE
Baboon (Papio (142+3.5 (3.8+0.5 (107+3. (2.26+0.4 (2.10+0.0
sp)c ) ) 7) 8) 2)
Cynomolgus 142-153f 3.0-4.8f 101-112f 1.18-2.30g 2.17-
(M. fascicularis) (149) (3.9) 2.55g
(107) (2.36)
Rhesus 154-158 3.6-4.6 110-114 1.41-1.62 2.42-2.70 0.68-0.75
(M. mulatta)c
PIGb 135-152 4.4-6.7 94-106 18-27 1.71-3.10 1.78-2.90 1.11-1.52
(1.31+0.2
(2.41+0.2 0)
5)
RABBITb 85-105.3 1.46-3.60

(141+0.9 (5.30.5) (96.5+6. (47) (1.34+0.1 (0.92+0.0

3) 7) 5) 7)
RATd 141-150h 5.2-7.8h 99-114h 1.99-3.77f 2.67- 1.07-1.28c
Wistar[Crl: (6.2) (106) 3.43f
(W)BR] (145) (2.95) (3.05)
i
F-344 139-150 3.9-7.5 82-99 2.42-5.62 2.47-3.32
[CDF(F- (145) (5.7) (93) (4.13)
344)CrlBR] (2.82)
CD[Crl:CD(SD) 139-150 3.6-8.4 84-99 2.42-5.62 2.47-3.22 0.66-1.79c
BR]j (145) (5.7) (93) (4.13)
(2.82)
SHEEPb 139-152 3.9-5.8 95-103 20-25 1.62-2.36 2.88-3.20 0.90-0.31
(1.03+0.1
(2.78+0.0 2)
7)

a
Ranges with the means and standard deviations in parenthesis. Reported in S.I. units.
b
KANEKO, J.J., ed. Clinical chemistry of domestic animals. Academic Press, 1989: 886-
891.
c
LOEB, W.F. and QUIMBY, F.W., eds. The clinical chemistry of laboratory animals.
Pergamon Press, 1989: 417-476.

312
d
Sexes combined, 19-21 weeks.
e
Baseline haematology and clinical chemistry values for Charles River outbred mice:
Crl:CD-1(ICR)BR. Crl:CF-1BR. Charles River Laboratories Techn. Bull., 1986.
f
Values from Parke Davis Research Institute, Mississauga, Ontario.
g
CLARKE, D., TUPASI, G., WALKER, R. and SMITH, G. Stability of serum
biochemical parameters in beagle dogs and cynomolgus monkeys. Clin. Chem. Newsl.
(In press).
h
Baseline haematology and clinical chemistry values for Charles River Wistar rats (CRL:
(W)BR) as a function of sex and age. Charles River Techn. Bull., Vol. 1, No. 2, 1982.
i
Baseline haematology and clinical chemistry values for Charles River Fischer-344 rats -
CDF(F-344)CrlBR as a function of sex and age. Charles River Techn. Bull., Vol. 3, No. 1,
1984.
j
Baseline haematology and clinical chemistry values for Charles River
CD[Crl:CD(SD)BR] rats as a function of sex and age. Charles River Techn. Bull., Vol. 3,
No. 2, 1984.

APPENDIX VIII

COMMON BLEEDING SITES

SPECIES ANT, CEPHA EAR FEMOR HEAR JUGUL ORBIT TAI TOE/T WIN
VEN LIC VEIN AL T AR AL L AIL G
A VEIN VEIN VEIN SINUS V/A NICK VEI
CAV N
A
CAT Xc Xc,r,t Xc,r,t
COW Xj,t Xt
BIRD Xc,f Xc Xc,f
DOG Xc,r,t,u Xc Xc,r,t,u
FERRET Xl Xl Xl Xl
FISH Xe Xe
FROG Xg
GERBIL Xv Xv Xv

313
GUINEA Xaa Xn,o Xo Xo Xc,o
PIG
HAMSTE Xx Xp Xp,x Xx Xx
R
NHP Xc,w Xc
MOUSE Xc,h,i, Xc,h,n,s Xh,s Xc,i,n,s
n,s
OPOSSU Xk Xk
M
RABBIT Xa,c,b Xa,c,b
,n
RAT Xc,n,z, Xy Xc,n,z,y Xd,y Xc,n,z,y
y
SMALL Xt Xt
RUMINA
NT
SNAKE Xm
PIG Xq,t Xq,t
TURTLE Xm Xm

X Recognized bleeding site, followed by reference.

References
a
ADAMS, C.E. The laboratory rabbit. In: Poole, T.B., ed. UFAW (Universities
Federation for Animal Welfare) handbook on the care and management of laboratory
animals. 6th Ed. Harlow, Essex: Longman Scientific and Technical, 1987: 415-435.
b
BIVIN, W.S. et TIMMONS, E.H. Basic biomethodology. In: Weisbroth, S.H., Flatt, R.E.
and Kraus, A.L., eds. The biology of the laboratory rabbit. New York, NY: Academic
Press, 1974: 74-89.
c
BIVIN, W.S. et SMITH, G.D. Techniques of experimentation. In: Fox, J.G., Cohen, B.J.
and Loew, F.L., eds. Laboratory animal medicine. Orlando, FL: Academic Press, 1984:
564-588.
d
BOBER, R. Technical review: Drawing blood from the tail artery of a rat. Lab. Anim.
July/August, 1988: 33-34.
e
CAMPBELL, T.W. Fish cytology and hematology. In: Stoskopf, M.K., ed. The
Veterinary Clinics of North America. Small Animal Medicine. Tropical Fish Medicine.
Philadelphia, PA: W.B. Saunders, 1988: 349-364.

314
f
COOPER, D.M. et HARRY, E.G. The domestic fowl and turkey. In: Poole, T.B., ed.
UFAW (Universities Federation for Animal Welfare) handbook on the care and
management of laboratory animals. 6th Ed. Harlow, Essex: Longman Scientific and
Technical, 1987: 640-662.
g
CRAWSHAW, G.J. Amphibian medicine. In: Kirk, R.W. and Bonagura, J.D., eds.
Current veterinary therapy XI small animal practice. Philadelphia, PA: W.B. Saunders,
1992: 1219-1230.
h
CUNLIFFE-BEAMER, T.L. Biomethodology and surgical techniques. In: Foster, H.L.,
Small, J.D. and Fox, J.G., eds. The mouse in biomedical research. Vol. III. Normative
biology, immunology, and husbandry. New York, NY: Academic Press, 1983: 402-439.
i
CUNLIFFE-BEAMER, T.L. et LES, E.P. The laboratory mouse. In: Poole, T.B., ed.
UFAW (Universities Federation for Animal Welfare) handbook on the care and
management of laboratory animals. 6th Ed. Harlow, Essex: Longman Scientific and
Technical, 1987: 275-308.
j
EWBANK, T. Cattle. In: Poole, T.B., ed. UFAW (Universities Federation for Animal
Welfare) handbook on the care and management of laboratory animals. 6th Ed. Harlow,
Essex: Longman Scientific and Technical, 1987: 525-534.
k
FINNIE, E.P. Monotremes and marsupials. Restraint. In: Fowler, M.E., ed. Zoo and wild
animal medicine. Philadelphia, PA: W.B. Saunders, 1986: 181-184.
l
FOX, J.G. Anesthesia and surgery. In: Fox, J.G., ed. Biology and diseases of the ferret.
Philadelphia, PA: Lea and Febiger, 1988: 289-302.
m
FRYE, F.L. Hematology of captive reptiles. In: Fowler, M.E., ed. Zoo and wild animal
medicine. Philadelphia, PA: W.B. Saunders, 1986: 181-184.
n
HARKNESS, J.E. et WAGNER, J.E. Clinical procedures. In: The biology and medicine
of rabbits and rodents. Philadelphia, PA: Lea and Febiger, 1989: 55-76.
o
HOAR, R.M. Biomethodology. In: Wagner, J.E. and Manning, P.J., eds. The biology of
the guinea pig. New York, NY: Academic Press, 1976: 13-17.
p
HOBBS, K.R. Hamsters. In: Poole, T.B., ed. UFAW (Universities Federation for Animal
Welfare) handbook on the care and management of laboratory animals. 6th Ed. Harlow,
Essex: Longman Scientific and Technical, 1987: 377-410.
q
HOLTZ, W. Pigs and minipigs. In: Poole, T.B., ed. UFAW (Universities Federation for
Animal Welfare) handbook on the care and management of laboratory animals. 6th Ed.
Harlow, Essex: Longman Scientific and Technical, 1987: 496-515.

315
r
KIRK, R.W., BISTNER, S.I. et FORD, R.B., eds. Routine diagnostic procedures. In:
Handbook of veterinary procedures and emergency treatment. 5th Ed. Philadelphia, PA:
W.B. Saunders, 1990: 447-455.
s
KRAUS, A.L. Research methodology. In: Baker, H.J., Lindsey, J.R. and Weisbroth,
S.H., eds. The laboratory rat. Vol. II. Research applications. New York, NY: Academic
Press, 1980: 5-10.
t
LUCAS, M.J. et LUCAS, S.E. Diagnostic sampling and treatment techniques. In:
McCurrin, D.M., ed. Clinical Textbook for Veterinary Technicians. 2nd Ed. Philadelphia,
PA: W.B. Saunders, 1990: 191-209.
u
MACARTHUR, J.A. The dog. In: Poole, T.B., ed. UFAW (Universities Federation for
Animal Welfare) handbook on the care and management of laboratory animals. 6th Ed.
Harlow, Essex: Longman Scientific and Technical, 1987: 456-473.
v
NORRIS, M.L. Gerbils. In: Poole, T.B., ed. UFAW (Universities Federation for Animal
Welfare) handbook on the care and management of laboratory animals. 6th Ed. Harlow,
Essex: Longman Scientific and Technical, 1987: 360-376.
w
OLSON, M.E., MORCK, D.W. et NABROTZKY, V.C.A. Manual of standard operating
procedures for animal facilities. Calgary, Alberta: University of Calgary, 1992.
x
SILVERMAN, J. Biomethodology. In: Van Hoosier, G.L. and McPherson, C.W., eds.
Laboratory hamsters. Orlando, FL: Academic Press, 1987: 7-91.
y
WAYNFORTH, H.B. et FLECKNELL, P.A. Methods of obtaining body fluids. In:
Experimental and surgical techniques in the rat. 2nd Ed. London, U.K.: Academic Press,
1992: 68-87.
z
WEIHE, W.B. The laboratory rat. In: Poole, T.B., ed. UFAW (Universities Federation
for Animal Welfare) handbook on the care and management of laboratory animals. 6th
Ed. Harlow, Essex: Longman Scientific and Technical, 1987: 309-330.
aa
WHORTON, J.A. Collecting blood via the anterior vena cava in the guinea pig. Lab
Animal 1982; 11(6): 66-68.

316
 APPENDIX IX

TRANQUILLIZER, SEDATIVE AND ANTICHOLINERGIC DRUG DOSAGES

Acepromazine Xylazine Midazola Diazepam Atropine Glycopyrrolatec

Maleate (Rompu ma a
Sulfatec
d
(Atravet) m) (Versed) (Valium)
SPECIE Mg/ Routeb Mg/ Rou Mg/ Rou Mg/ Rou Mg/ Rou Mg/ Route
S Kg Kg te Kg te Kg te Kg te Kg
CAT 0.2- IM IV 1-3 IM 0.2- IM 1.0- 0.02- SC 0.011 IM
0.5 PO SC 0.5 IV max IM IV
1-3 5 mg 0.05 IV 0.005
CATTLE 0.1 IV 0.1 IM not
effective
DOG 0.1- IM IV 1-2 IM 0.2- IM 1.0 IM 0.02- SC 0.011 IM
0.5 SC 0.5 IV max IV IM IV
20.0 0.05 IV 0.005
GUINEA 5.0 IP 2.5 IM 0.02- SC
IP IM
PIG 0.05 IV
HAMST 5.0 IP 5.0 IP 0.02- SC
ER/ 5.0 IP IM
GERBIL 0.05 IV
MOUSE 5.0 IP 1.0 IM 0.1- SC
IV 0.2
IM
IV
NON- 0.5-1 SC IM 1-2 IM 1.0 IM 0.05 SC
HUMAN IV IM
IV
PRIMAT
E
RABBIT 1.0 SC IM 1-3.0 IM 2.0 IP 1.0 IM 0.1- SC 0.1 SC
IV 0.2 IM
IV
RAT 2.5 IP 2.5 IP 0.02- SC
IM
0.05 IV
SHEEP/ 0.1- IM IV 1.0 IM 0.05 SC
GOAT 0.2 0.05 IM IM
SWINE 0.2 IM IV 1-2 IM 0.05- SC

317
 IV IM
0.1 IV

a
The tranquillizers listed are not marketed in Canada under veterinary labels. These are
human Schedule F, Part II (H&W Canada)
drugs requiring prescription.
b
PO = oral; SC = subcutaneous; IM = intramuscular; IP = intraperitoneal; IV =
intravenous.
c
Atropine and Glycopyrrolate should be administered 35-50 minutes prior to surgery, SC
or IM.
d
Xylazine--is an analgesic as well as a sedative.

APPENDIX X

ANALGESIC DRUG DOSAGES

SPECI Acetylsalicy Meperidine Fentanyl + Morphine Butorphano Buprenorph

ES lic Acid HCI Droperidol** l ine
(Aspirin) (Pethidine)
(Demerol) (Innovar-
vet)*
Mg/ Durati Mg/ Durati ml/k Duratio Mg/ Durati Mg/ Durati Mg/ Durati
Kg on Kg on g n Kg on Kg on Kg on
Rout Rout Rout Rout Rout Rout
e e e e e e
CAT 2-6 2-3 hr 0.05- 4 hr 0.4 3-4 hr 0.005 8-12 hr
IM 0.1 SC -0.01
SC SC SC
IV
CATTL
E
DOG 10 8-12 2-6 1-2 hr 0.2- 0.3- 2-4 hr 0.2- 3-4 hr 0.01- 8-12 hr
PO hr SC 0.5 2.0 0.4 0.02
IM IM SC SC IM
IM IM SC
IV
GOAT up to 4 hr up to 4 hr 0.005 8-12 hr
200 10 SC
total total IM
dose dose
IM

318
 IM
GUINE 85 4 hr 10- 2-3 hr 2-5 2-4 hr 0.05 6-12 hr
A PIG PO 20 SC SC
SC
IM
HAMST 0.02- 0.5 6-8 hr
ER 0.05 SC
ml/
100 g
IM
0.1 of
1:10
diluti
on IP
MOUSE 120- 4 hr 10- 2-3 hr 0.02- 2-5 2-4 hr 1-5 4 hr 0.05- 6-8 hr
300 20 0.05 SC SC 0.1
PO SC ml/ SC
IM 100 g
IM
NON- 10- 6 hr 2-4 3-4 hr 0.05- required 1-2 4 hr 0.025 4 hr 0.01- 8-12 hr
HUMA 20 IM 0.2 dosages SC IM 0.05
N PO IM vary IM IM
PRIMA greatly IV
TE with
differen
t NHP
species
RABBI 10 4 hr 10- 2 hr 0.15- 2-5 2-4 hr 0.1- 4 hr 0.02- 8-12 hr
T PO 20 2-4 hr 0.3 SC 0.5 0.05
SC IM IM IV SC
IM IV
5 IV IM
RAT 100 4 hr 10- 2-3 hr 0.10- Sedatio 2-5 2-4 hr 2 SC 4 hr 0.01- 8-12 hr
PO 20 0.25 n/ SC 0.5
SC IM Anesthe SC
IM 0.2- sia.
0.5 Dilute
IM to 10%
solution
prior to
adminis
tra-
tion
SHEEP up to 4 hr up to 4 hr 0.005 4-6 hr

319
 200 10 IM
total total
dose dose
IM IM
SWINE 10 4 hr 2 IM 4 hr 0.5 up to 4 hr 0.1- 4 hr 0.1 8-12 hr
PO IM 10 0.3 IV
0.03 total IM IM
IV dose
IM

* Innovar-vet = Fentanyl 0.4 mg/ml + Droperidol 20 mg/ml. ** Neurolepranalgesic.

References

Anesthesia and analgesia in laboratory animals. In: Proc. American College of

Laboratory Animal Medicine (ACLAM) Forum, 1990.

FLECKNELL, P. Laboratory animal anesthesia. Orlando, FL: Academic Press, 1987.

FLECKNELL, P. Anesthesia of laboratory animals. Short course. University of Guelph,

Guelph, Ontario 1992.

APPENDIX XI

INJECTABLE ANESTHETIC AGENTS--DOSAGE

SPECIE Pentobarbital Thiopenta Ketamine Urethaneb Ketamine/ Alphaxalone/

S l HCIa Alphadolone
Xylazine (Saffan)c
Mg/K Route Mg/K Rout Mg/K Rout Mg/K Rout Mg/K Rout Mg/K Route
g g e g e g e g e g
CAT 25 IV 10-15 IV 20 IM 1250 IV 15/ IM 9-12 IV
1 IM 12-18 IM

320
 SC
DOG 20-30 IV 25 IV 1000 IV contraindicated
GOAT 30 IV 15 IV 20 IM
GUINEA 37 IP 20 IV 100- IM 1500 IP 40- IM 40 IP
200 IV 100/ IM
4-5 SQ
PIG
HAMST 50-90 IP 20-40 IV 150 IP
ER IP
MOUSE 30-40 IP 30-40 IV 100- IM 200/ IM 10-15 IV
IP 200 10 IP
NON- 5-15 IV 15-20 IV 5-25 IM 7/ IM 6-9 IV
HUMAN 0.6 IM 12-18 IM

PRIMAT
E
RABBIT 45 IV 20 IV 50 IM 1000 IV 35- IM 6-9 IV
IP 50/ IM
5-10
RAT 40 IP 20-40 IV 60- IM 1000 IP 90/ IP 10-12 IV
IP 100 5-10 IM
IP
IM
SHEEP 30 IV 15 IV 20 IM
SWINE 30 IV 6-8 IV 10 IM 20/ IM 2 IV
2 IM 5 IM

a
Ketamine useful for birds at 15-20 mg/kg for immobilization and from 40-100 mg/kg
for anesthesia in healthy birds, alone or in combination with a suitable tranquillizer.
b
May only be used for non-survival surgery--gives prolonged anesthesia. CAUTION:
Urethane is carcinogenic.
c
Saffan useful as anesthetic for birds given IV rapidly at 12-14 mg/kg body weight.

APPENDIX XIV

METHODS FOR EUTHANASIA BY SPECIES

(Methods in order of Acceptability)

321
 Species Most Acceptable Acceptable
AMPHIBIANS Barbiturates Double pithing
Inhalant anesthetics Decapitation and
Tricaine methanesulfonate MS-222 pithing
Benzocaine Stunning and
pithing
CO2 + O2
mixture
AVIAN SPECIES Barbiturates Electrocution
(birds) Inhalant anesthetics stunning
followed by
exsanguination

CO2 + O2
mixture
Physical
stunning
followed by
exsanguination
or
decapitation
BOVINE SPECIES Barbiturates Shooting
(calves, cows, goats, sheep Penetrating captive bolt followed by followed by
and other ruminants) exsanguination
exsanguination
CATS Inhalant anesthetics CO2 + O2
Barbiturates mixture
DOGS Inhalant anesthetics CO2 + O2
Barbiturates mixture
EQUINES (horses) Barbiturates Shooting
Penetrating
captive bolt
FISH Tricaine methanesulfonate MS-222 Stunning
Benzocaine followed by
cervical
dislocation or
decapitation
FUR BEARING Barbiturates CO
ANIMALS Electrical stunning using special CO2 + O2
(mink, fox, others raised equipment followed by cervical
for fur) dislocation
Inhalant anesthetics in specially
designed chamber

322
 INVERTEBRATES Tricaine methanesulfonate MS-222 CO2 bubbling
(cephalopods, crustacea) Benzocaine through water
MARINE MAMMALS Barbiturates Stunning
(seals, porpoises, cetacea) Etorphine hydrochloride followed by
exsanguination
NON-HUMAN Barbiturates Tranquillization
PRIMATES Inhalant anesthetics and CO2 + O2
mixture
RABBITS Barbiturates CO2 + O2
Inhalant anesthetics mixture
REPTILES Inhalant anesthetics in special CO2 + O2
chamber mixture
Barbiturates
RODENTS (and similar Inhalant anesthetics in special ' CO2 + O2
small species) chambers mixture
Barbiturates CO
Microwave irradiation in specially
designed units
SWINE Barbiturates Electrocution
Inhalant anesthetics using special
equipment
WILD AMIMALS Shooting by expert marksman Sedation
(200 species, etc.) Immobilization followed by followed by
barbiturates penetrating
captive bolt

Barbiturates includes an overdose of all pharmacological agents which depress the

Central Nervous System (CNS) producing irreversible unconsciousness and death (see
also Euthanasia and CCAC policy statements).

Competence and knowledge of agents and procedures are mandatory requirements for
personnel conducting euthanasia.

Any other means (not listed here) of killing an experimental animal, must not be
undertaken without prior review and approval by an Animal Care Committee (ACC), and
should only be done in expert hands.

APPENDIX XV-A

ETHICS OF ANIMAL INVESTIGATION

323
The use of animals in research, teaching, and testing is acceptable only if it promises to
contribute to understanding of fundamental biological principles, or to the development
of knowledge that can reasonably be expected to benefit humans or animals.

Animals should be used only if the researcher's best efforts to find an alternative have
failed. A continuing sharing of knowledge, review of the literature, and adherence to the
Russell-Burch '3R' tenet of 'Replacement, Reduction and Refinement' are also requisites.
Those using animals should employ the most humane methods on the smallest number of
appropriate animals required to obtain valid information.

The following principles incorporate suggestions from members of both the scientific and
animal welfare communities, as well as the organizations represented on Council. They
should be applied in conjunction with the Canadian Council on Animal Care's (CCAC)
'Guide to the Care and Use of Experimental Animals.'

1. If animals must be used, they should be maintained in a manner that provides for their
physical comfort and psychological well-being, according to CCAC's 'Policy Statement
on Social and Behavioural Requirements of Experimental Animals.'

2. Animals must not be subjected to unnecessary pain or distress. The experimental

design must offer them every practicable safeguard, whether in research, in teaching or in
testing procedures; cost and convenience must not take precedence over the animal's
physical and mental well-being.

3. Expert opinion must attest to the potential value of studies with animals. The following
procedures, which are restricted, require independent, external evaluation to justify their
use:

i) burns, freezing injuries, fractures, and other types of trauma investigation in

anesthetized animals,
concomitant to which must be acceptable veterinary practices for the relief of pain,
including adequate
analgesia during the recovery period;

ii) staged encounters between predator and prey or between conspecifics where
prolonged fighting and injury
are probable.

4. If pain or distress is a necessary concomitant to the study, it must be minimized both in

intensity and duration. Investigators, Animal Care Committees (ACC), grant review
committees and referees must be especially cautious in evaluating the proposed use of the
following procedures:

a) experiments involving withholding pre- and post-operative pain-relieving medication;

324
b) paralyzing and immobilizing experiments where there is no reduction in the sensation
of pain;

c) electric shock as negative reinforcement;

d) extreme environmental conditions such as low or high temperatures, high humidity,

modified atmospheres,
etc., or sudden changes therein;

e) experiments studying stress and pain;

f) experiments requiring withholding of food and water for periods incompatible with the
species specific
physiological needs; such experiments should have no detrimental effect on the health
of the animal;

g) injection of Freund's Complete Adjuvant (FCA). This must be carried out in

accordance with 'CCAC
Guidelines on Acceptable Immunological Procedures.'

5. An animal observed to be experiencing severe, unrelievable pain or discomfort, should

immediately be
humanely killed, using a method providing initial rapid unconsciousness.

6. While non-recovery procedures involving anesthetized animals, and studies involving

no pain or distress are considered acceptable, the following experimental procedures
inflict excessive pain and are thus unacceptable:

a) utilization of muscle relaxants or paralytics (curare and curare-like) alone, without

anesthetics, during surgical
procedures;

b) traumatizing procedures involving crushing, burning, striking or beating in

unanesthetized animals.

7. Studies such as toxicological and biological testing, cancer research and infectious
disease investigation may, in the past, have required continuation until the death of the
animal. However, in the face of distinct signs that such processes are causing irreversible
pain or distress, alternative endpoints should be sought to satisfy both the requirements of
the study and the needs of the animal.

8. Physical restraint should only be used after alternative procedures have been fully
considered and found inadequate. Animals so restrained, must receive exceptional care
and attention, in compliance with species specific and general requirements as set forth in
the 'Guide.'

325
9. Painful experiments or multiple invasive procedures on an individual animal,
conducted solely for the instruction of students in the classroom, or for the demonstration
of established scientific knowledge, cannot be justified. Audiovisual or other alternative
techniques should be employed to convey such information.

Revised October 1989

APPENDIX XV-B

CATEGORIES OF INVASIVENESS IN ANIMAL EXPERIMENTS

Investigators and teachers who consider it essential to use vertebrates or invertebrates in

their research, teaching or testing in the laboratory or in the field, must adhere to
humane principles, and take cognizance of the Canadian Council on Animal Care's
(CCAC) Ethics of Animal Investigation and other CCAC documentation in assigning a
category. Protocols must be submitted to an appropriate review committee for all studies
and courses which involve the use of vertebrates and some invertebrates in Categories B
through E. Cephalopods and some other higher invertebrates have nervous systems as
well developed as in some vertebrates, and may therefore warrant inclusion in Category
B, C, D, or E.

The following list of categories provides possible examples of experimental procedures

which are considered to be representative of each category.

A. Experiments on most invertebrates or on live isolates

Possible examples: the use of tissue culture and tissues obtained at necropsy or from the
slaughterhouse; the use of eggs, protozoa or other single-celled organisms; experiments
involving containment, incision or other invasive procedures on metazoa.

B. Experiments which cause little or no discomfort or stress

Possible examples: domestic flocks or herds being maintained in simulated or actual

commercial production management systems; the short-term and skillful restraint of
animals for purposes of observation or physical examination; blood sampling; injection
of material in amounts that will not cause adverse reactions by the following routes:
intravenous, subcutaneous, intramuscular, intraperitoneal, or oral, but not intrathoracic or
intracardiac (Category C); acute non-survival studies in which the animals are completely
anesthetized and do not regain consciousness; approved methods of euthanasia following
rapid unconsciousness, such as anesthetic overdose, or decapitation preceded by sedation
or light anesthesia; short periods of food and/or water deprivation equivalent to periods of
abstinence in nature.

326
C. Experiments which cause minor stress or pain of short duration

Possible examples: cannulation or catheterization of blood vessels or body cavities under

anesthesia; minor surgical procedures under anesthesia, such as biopsies, laparoscopy;
short periods of restraint beyond that for simple observation or examination, but
consistent with minimal distress; short periods of food and/or water deprivation which
exceed periods of abstinence in nature; behavioural experiments on conscious animals
that involve short-term, stressful restraint; exposure to non-lethal levels of drugs or
chemicals. Such procedures should not cause significant changes in the animal's
appearance, in physiological parameters such as respiratory or cardiac rate, or fecal or
urinary output, or in social responses.

Note: During or after Category C studies, animals must not show self-mutilation,
anorexia, dehydration, hyperactivity, increased recumbency or dormancy, increased
vocalization, aggressive-defensive behaviour or demonstrate social withdrawal and self-
isolation.

D. Experiments which cause moderate to severe distress or discomfort

Possible examples: major surgical procedures conducted under general anesthesia, with
subsequent recovery; prolonged (several hours or more) periods of physical restraint;
induction of behavioural stresses such as maternal deprivation, aggression, predator-prey
interactions; procedures which cause severe, persistent or irreversible disruption of
sensorimotor organization; the use of Freund's Complete Adjuvant (FCA) (see CCAC
Guidelines on Acceptable Immunological Procedures).

Other examples include induction of anatomical and physiological abnormalities that will
result in pain or distress; the exposure of an animal to noxious stimuli from which escape
is impossible; the production of radiation sickness; exposure to drugs or chemicals at
levels that impair physiological systems.

Note: Procedures used in Category D studies should not cause prolonged or severe
clinical distress as may be exhibited by a wide range of clinical signs, such as marked
abnormalities in behavioural patterns or attitudes, the absence of grooming, dehydration,
abnormal vocalization, prolonged anorexia, circulatory collapse, extreme lethargy or
disinclination to move, and clinical signs of severe or advanced local or systemic
infection, etc.

E. Procedures which cause severe pain near, at, or above the pain tolerance
threshold of unanesthetized conscious animals

This Category of Invasiveness is not necessarily confined to surgical procedures, but may
include exposure to noxious stimuli or agents whose effects are unknown; exposure to
drugs or chemicals at levels that (may) markedly impair physiological systems and which
cause death, severe pain, or extreme distress; completely new biomedical experiments
which have a high degree of invasiveness; behavioural studies about which the effects of

327
the degree of distress are not known; use of muscle relaxants or paralytic drugs without
anesthetics; burn or trauma infliction on unanesthetized animals; a euthanasia method not
approved by the CCAC; any procedures (e.g., the injection of noxious agents or the
induction of severe stress or shock) that will result in pain which approaches the pain
tolerance threshold and cannot be relieved by analgesia (e.g., when toxicity testing and
experimentally-induced infectious disease studies have death as the endpoint).

Revised February 1991

APPENDIX XV-C

CCAC GUIDELINES ON ACCEPTABLE IMMUNOLOGICAL

PROCEDURES

When beginning an immunization, choosing the correct adjuvant may be difficult. As a

general suggestion, Freund's Complete Adjuvant (FCA) may be used when only small
amounts of soluble immunogens are available. FCA is considered to be an emulsion
consisting of equal volumes of FCA to antigen (1 part FCA or less to 1 part antigen). If
large amounts of particulate, or highly immunogenic immunogens are available, other
adjuvants should be considered.

An important aspect in immunization procedures is the utilization of skilled, competent,

technical staff experienced in the handling of the species being used and in performing
the technique. They must be knowledgeable and capable of recognizing signs of distress
in all injected animals, and be responsible for taking action when necessary.

FCA should be used only for the most problematic immunization situations. It must
never be given either intravenously or in repeated doses. FCA must not be used in
horses.

Intradermal Route

Sound scientific evidence and justification must be available if the intradermal route of
injection of FCA is to be used, because of the frequent ulceration and infections that
occur at the site of such injections. The use of the intradermal route may be justified only
when the purpose is to induce cell-mediated response.

In rabbits, volumes of inoculum in excess of 0.05 mls (50 microliters) per site should not
be used. The location of the site(s) should be carefully selected so as to prevent
mutilation. A minimal number of sites should be selected, and the distance between each
site be maximized.

The intradermal route is inappropriate in the mouse. Nor is it recommended in other

328
rodents.

Subcutaneous Route

In guinea pigs, up to a total volume of 0.4 ml (400 microliters) of inoculum may be

injected subcutaneously dorsally in the neck, in one or divided into several sites. In
rabbits, the site of choice is the interscapular region (between the shoulder blades) on the
dorsum (back), administering up to 0.25 ml of inoculum (250 microliters) per site, to a
maximum of four sites. The distance between sites should be maximized. In the mouse,
up to 0.1 ml (100 microliters) may be administered in the neck region.

Intramuscular Route

In rabbits, intramuscular injections of FCA may be administered in the thigh muscle; up

to 0.5 ml (500 microliters), preferably in one site. Intramuscular injection of FCA is not
recommended for small laboratory animals such as rats, mice, hamsters, gerbils, etc.. For
larger animals such as cats, dogs and poultry, up to 1 ml of FCA injected into the thigh
muscles is acceptable. In livestock such as pigs, cattle, sheep and goats, the intramuscular
route is acceptable.

Intraperitoneal Route

The intraperitoneal route for injection of FCA is permitted in small rodents only. FCA
should be administered only once, and be limited to minimal volumes of up to 0.1 ml
(100 microliters).

Intravenous Route

FCA is not to be injected intravenously.

Footpad Injection

FCA should not be injected in the feet of rabbits. Footpad injection of FCA in rodents is
not permissible unless there is scientific evidence indicating this route is essential as a
specific requirement for the production of immune response. In rats and mice, only one
footpad may be used. Animals should be maintained on soft bedding and not on wire-
bottomed cages.

Induction of Ascites Fluid in Animals

Pristane or other recognized priming agent(s) (excluding FCA) may be used.

Ascites may be collected only for as long as the animal is not experiencing pain or
distress, is in good body condition, and does not show signs of debilitation, dehydration
or other complications from the procedure. Upon recognition of loss of condition, pain, or
distress the animal must be euthanized according to a method approved by the Canadian

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Council on Animal Care (CCAC).

Observation of Injection Sites

The injection site(s) must be observed by the investigator or his/her designate, a

minimum of three times per week, for four weeks after each injection.

If a lesion(s) develops at any injection site, it must be reported through established

channels, e.g., the animal resources supervisor or veterinarian, and must receive
appropriate veterinary treatment. Such lesions should be inspected at least three times per
week by the investigator or his/her designate, until all lesions are healed.

Related Links

The following are links to websites which may be useful to visitors to the Canadian
Council on Animal Care (CCAC) website. This page will be expanded and updated
regularly. If you are aware of another link which you feel should be included in this list,
or if you have found that anyone of the links are no longer active, please contact the
CCAC at ( [email protected] ).

Canadian Organizations

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 AAC - Animal Alliance of Canada

AAFC - Agriculture and Agri-Food Canada (AAFC)

ACFD - Association of Canadian Faculties of Dentistry

AFMC - Association of Faculties of Medicine of Canada

AUCC - Association of Universities and Colleges of Canada

CAHI - Canadian Animal Health Institute

CALAM - Canadian Association for Laboratory Animal Medicine

CALAS - Canadian Association for Laboratory Animal Science

CAZA - Canadian Association of Zoos and Aquariums

CAZWV - Canadian Association of Zoo and Wildlife Veterinarians

CCFAVM - Confederation of Canadian Faculties of Agriculture and Veterinary Medicine

CBS - Canadian Bioethics Society

CFBS - Canadian Federation of Biological Societies

CFHS - Canadian Federation of Humane Societies

CIHR - Canadian Institutes of Health Research

CMA - Canadian Medical Association

CSZ - Canadian Society of Zoologists

CVMA - Canadian Veterinary Medical Association

DFO - Department of Fisheries and Oceans Canada

DND - Department of National Defence

EC - Environment Canada

HC - Health Canada

HSFC - Heart and Stroke Foundation of Canada

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 NCIC - National Cancer Institute of Canada

NRC - National Research Council

NSERC - Natural Sciences and Engineering Research Council

RSC - The Royal Society of Canada

Rx&D - Canada's Research-Based Pharmaceutical Companies

Canadian Guidelines/Legislation

Codes of Practice and Factsheets for the Care and Handling of Farm Animals, Canadian
Agri-Food Research Council (CARC)

Health Canada - Laboratory Biosafety Guidelines(2004)

Schedule III, Tri-Agency (CIHR, NSERC & SSHRC) Memorandum of understanding on

the Roles and Responsibilities in the Management of Federal Grants and Awards (MOU)

Canadian Universities

Simon Fraser University, Research Policies

University of Calgary, Research Policy-Animal Care

University of Guelph, Animal Care Services

University of Manitoba, Research Policies, Guidelines and Procedures

University of Northern British Columbia, Policy and Procedures on Animal Care

US & International Organizations

AAALAC - Association for Assessment and Accreditation of Laboratory Animal Care

International

AALAS - American Association for Laboratory Animal Science

ACLAM - American College of Laboratory Animal Medicine

AFIP - Armed Forces Institute of Pathology Department of Veterinary Pathology

ANZCCART - Australian and New Zealand Council for the Care of Animals in Research
and Teaching

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 APHIS - Animal and Plant Health Inspection Service (US)

APS - American Physiological Society

ASLAP - American Association of Laboratory Animal Practitioners

ASPCA - American Society for the Prevention of Cruelty to Animals

AWIC - Animal Welfare Information Center

AVMA - American Veterinary Medical Association

CAAT - Center for Alternatives to Animal Testing

CDC - Centers for Disease Control

ECVAM - European Centre for the Validation of Alternative Methods

FBR - Foundation for Biomedical Research

FRAME - Fund for the Replacement of Animals in Medical Experiments

ICLAS - International Council for Laboratory Animal Science

IICA - Inter-American Institute for Cooperation on Agriculture

ILAR - Institute of Laboratory Animal Resources

LASA - Laboratory Animal Science Association

MISMR - Michigan Society for Medical Research

NABR - National Association for Biomedical Research

NCRR - National Center for Research Resources (NIH-USA)

OHRP - Office for Human Research Protections (US)

PHS - Public Health Service

PRIM&R - Public Responsibility in Medicine and Research

RDS - Understanding Animal Research in Medicine

SCAW - Scientists Center for Animal Welfare

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 UFAW - Universities Federation for Animal Welfare

USDA - United States Department of Agriculture

US Guidelines/Legislation

APA - Guidelines for Ethical Conduct in the Care and Use of Animals

Institutional Animal Care and Use Committees (AWIC Document)

US Animal Welfare Act

US Guide to the Care and Use of Laboratory Animals

US Government Principles for the Utilization and Care of Vertebrate Animals Used in
Testing, Research and Training

European Guidelines/Legislation

Animals (Scientific Procedures) Act 1986 (United Kingdom)

Introduction to Norwegian Legislation on the Use of Laboratory Animals (Norway)

Lega antivivisezione (LAV) (Italian)

Reference Centres and Databases

ALTWEB

Collection of abstracts and bibliographic references to worldwide biological and medical

literature

ERGATT FRAME ECVAM Databank of In Vitro Techniques in Toxicology

Federal Register

MEDWEB

MICROBES.INFO - The Microbiology Information Portal

National Centre for the Replacement, Refinement and Reduction of Animals in Research

NETVET

NORINA - Norwegian Inventory of Alternatives - Audiovisual Alternatives to Laboratory

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 Animals in Teaching
Norwegian Reference Centre for Laboratory Animal Science and Alternatives

Primate Enrichment Database

References: The Ethics of Using Transgenic Animals

References for Users of Transgenic Animals

Useful Websites Compiled for American Institutional Animal Care and Use Committees
(IACUCs)

Alternatives

AWIC - Animal Welfare Information Center

ALTWEB

ECVAM - European Centre for the Validation of Alternative Methods

ERGATT FRAME ECVAM Databank of In Vitro Techniques in Toxicology

FRAME - Fund for the Replacement of Animals in Medical Experiments

NORINA - Norwegian Inventory of Alternatives - Audiovisual Alternatives to Laboratory

Animals in Teaching
Norwegian Reference Centre for Laboratory Animal Science and Alternatives

Netherland Centre Alternatives (NCA)

Animal Welfare Sites

The Animal Welfare Institute

Educational Sites

The American Psychological Society (APS) and the Humane Use of Animals in Research

Animals in Research Questions and Answers (APS)

Blood collection in mice using the saphenous vein-an alternative to retro-orbital

collection

Brain Briefings: topics on neurosciences for the lay reader

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 Partners in Research

Virtual tour of an animal facility: Norwegian College of Veterinary Medicine

Journals/Newsletters

ILAR - Institute for Laboratory Animal Research

Lab Animal

Laboratory Animals

Laboratory Primate Newsletter

List of Online Veterinary Journals

Nature

New Scientist

Science

The Scientist

The Shape of Enrichment

Libraries

Bibliothque de mdecine vtrinaire de l'Universit de Montral

CISTI - Canada Institute for Scientific and Technical Information

The Electronic Zoo

Livestock Virtual Library at Oklahoma State University

National Agricultural Library

National Library of Medicine - PUBMED

Virtual Library Biosciences

Virtual Library Veterinary Medicine

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LIST OF ABBREVIATIONS

Espaol English Franais

Asociacin Canadiense de Canadian Association for Association canadienne
Ciencias de Animales de Laboratory Animal pour la science des
Laboratorio (ACCAL) Science (CALAS) animaux de laboratoire
(ACSAL)
Asociacin Canadiense de Association of Canadian
Ciencias de Animales de Medical Colleges Association des facults de
Laboratorio (ACCAL) (ACMC) mdecine du Canada
(AFMC)
Asociacin Canadiense de Association of Canadian
Facultades de Medicina Faculties of Dentistry Association des facults
(ACFM) (ACFD) dentaires du Canada
(AFDC)
Asociacin Canadiense de Canadian Food Inspection
Facultades de Odontologa Agency (CFIA) Agence canadienne
(ACFO) d'inspection des aliments
Canadian Association for (ACIA)
Agencia Canadiense de Laboratory Animal
Inspeccin de Alimentos Medicine (CALAM) Association canadienne
(ACIA) pour la mdecine des
Canadian Veterinary animaux de laboratoire
Asociacin Canadiense de Medical Association (ACMAL)
Medicina de Animales de (CVMA)
Laboratorio (ACMAL) Association canadienne
Canadian Psychological des mdecins vtrinaires
Asociacin Canadiense de Association (CPA) (ACMV)
los Medicos Veterinarios
(ACMV) Pharmaceutical Socit canadienne de
Manufacturers' psychologie (SCP)
Asociacin Canadiense de Association of Canada
Psicologa (ACP) (PMAC) Association canadienne de
l'industrie du mdicament
Asociacin de la Industria International Air (ACIM)
Farmacutica de Canad Transportation Association
(AIFC) (IATA) Association de transport
arien internationale
Asociacin International World Veterinary (ATAI)
de Transporte Areo Association (WVA)
(AITA) Association mondiale
Association of vtrinaire (AMV)
Asociacin Mundial Universities and Colleges
Veterinaria (AMV) of Canada (AUCC) Association des universits
et Collges du Canada

337
Asociacin de Atomic Energy Control (AUCC)
Universidades y Colegios Board (AECB) of Canada
de Canad (AUCC) Comission de contrle de
Canadian Council of l'energie atomique (CCEA)
Comisin de Control de la Departments of du Canada
Energa Atmica (CCEA) Psychology (CCDP)
del Canad Conseil canadien des
Canadian Council on dpartements de
Consejo Canadiense de Animal Care (CCAC) psychologie (CDDP)
Facultades de Psicologa
(CCFP) Confederation of Conseil canadien de
Canadian Faculties of protection des animaux
Consejo Canadiense de Agriculture and Veterinary (CCPA)
Proteccin de los Medicine (CCFAVM)
Animales (CCPA) Confdration des facults
Medical Research Council d'agriculture et de
Confederacin de (MRC) mdecine vtrinaire du
Facultades Canadienses de Canada (CFAMVC)
Agricultura y Medicina National Research
Veterinaria (CFCAMV) Council (NRC) Conseil de recherches
mdicales (CRM)
Consejo Mdico de Natural Sciences and
Investigacin (CMI) Engineering Research Conseil national de
Council (NSERC) recherches (CNR)
Consejo Nacional de
Investigacin (CNI) Heart and Stroke Conseil de recherches en
Foundation of Canada sciences naturelles et en
Consejo Nacional de (HSFC) gnie (CRSNG)
Investigacin en Ciencias
Naturales e Ingeniera Canadian Federation of Fondation des maladies du
(CNICNI) Biological Societies coeur du Canada (FMCC)
(CFBS)
Fundacin Canadiense de Fdration canadienne des
las Enfermedades del Canadian Federation of socits de biologie
Corazn (FCEC) Humane Societies (CFHS) (FCSB)

Federacin Canadiense de National Cancer Institute Fdration des socits

las Sociedades de Biologa (NCI) canadiennes d'assistance
(FCSB) aux animaux (FSCAA)
Agriculture and Agri-Food
Federacin de las Canada (AAFC) Institut national du cancer
Sociedades Canadienses (INC)
de Proteccin de los Department of National
Animales (FSCPA) Defense (DND) Agriculture et Agro-
alimentaire Canada (AAC)
Instituto Nacional del Environment Canada (EC)

338
Cncer (INC) Ministre de la Dfense
Fisheries and Oceans nationale (MDN)
Ministerio de Agricultura Canada (FOC)
y Agro-Alimentos de Environnement Canada
Canad (MAAAC) Health Canada (HC) (EC)

Ministerio de Defensa Canadian Bioethics Ministre des Pches et

Nacional (MDF) Society (CBS) Ocans (MPO)

Ministerio del Medio- Canadian Society of Sant Canada (SC)

Ambiente (MMA) Zoologists (CSZ)
Socit canadienne de
Ministerio de Ocanos y Royal Society of Canada biothique (SCB)
Pesca (MOP) (RSC)
Socit canadienne de
Ministerio de Salud de zoologie (SCZ)
Canad (MSC)
La Socit royale du
Sociedad Canadiense de Canada (SRC
Biotica (SCB)

Sociedad Canadiense de
Zoologa (SCZ)

Sociedad Real de Canad

(SRC)

339