To use animals properly for researchin Okayama University
by the Committee of the Animal Care and Use, Okayama University

Haruko Hirayama, DVM, PhD
Attending Veterinarian and Assistant Professor,
Department of Animal Resources,
Advanced Science Research Center, Okayama University

1

To use animals properly for research---

Animal experiments have been widely used as an
indispensable means of medical and life science research,and the results have contributed to the development of
medical technology and the overcoming of humandiseases. 
But,The ethical treatment of sacriﬁcial animals has attracted
attention, and even countries that have introduced stricter
animal experiment regulations than Japan are under strong
criticism. 
Therefore,
Introduce the concept of animal welfare in order tomaintain the framework for conducting necessary animal
experiments, comply with it, and maintain the current
situation as much as possible.
2

Responses to society required ofresearchers using animals-

Animal experiments can be conducted legally and sociallyas long as they are properly conducted for good reasons,but in reality, there are pros and cons to animal
experiments.

-

It is necessary to form a way of thinking about animal
experiments as a social norm that should be applied to all
people, to make it highly universal and objective, and toreach consensus among the people about theeﬀectiveness of animal experiments.

3

About this lecture
Purpose:To help ensure high-quality science andanimal well being
Object person:
All the researchers and the students
who are involved with use of living
animals for research and education
The term of validity:
Up to 5 ﬁnancial years

4Todayʼs contents
Law, Standards, andFundamental Guideline in Japan
University Policy and Regulation,and the other5

Law, Standards, andFundamental Guidelines in JapanAct on Welfare and Management of Animals- Law No. 105, 1973https://www.env.go.jp/nature/dobutsu/aigo/1̲law/ﬁles/aigo̲kanri̲1973̲105̲en.pdf

Standards Relating to the Care and Management of Laboratory Animals and Relief of Pain
- Notice No. 88 of the Ministry of the Environment, 2006
https://www.env.go.jp/nature/dobutsu/aigo/2̲data/laws/nt̲h25̲84̲en.pdf

Fundamental Guidelines for Proper Conduct of Animal Experimentand Related Activities in Academic Research Institutions- Notice No. 71 of the Ministry of Education, Culture, Sports,
Science and Technology, 2007
http://www.lifescience.mext.go.jp/policies/pdf/an̲material011.pdf
6

History of Humane Treatment of Animals
for Education and Research in Japan
1973“Act on Welfare and Management of Animals (Law No.
105)” was promulgated1970s Centralized animal facilities, like the main building of ourdepartment, were built in the medical schools, several nonmedical universities, and research institutions
1980s Institutional animal care and use committee (IACUC) started
to be organized, for the management of animal
experiments, including the review of animal use protocols
2005

Revision: The principles of the Three Rs was added, etc.

2012

Revision: “Lifelong breeding” was obligated, etc.

2019

Revision: Tightening regulations and penalties for animal
handling industry, etc.

7

The principles of the Three Rs
for humane experiments
What are the 3Rs?
First described by W. M. S. Russell and R. L. Burchin 1959 in their book,“The Principles of Humane Experimental Technique”
Replacement
Methods which avoid or replace the use of animals
Reduction
Methods which minimise the number of animals
used per experimentReﬁnement
Methods which minimise animal suﬀering andimprove welfare
Reference : NC3Rsʼs website : https://www.nc3rs.org.uk/the-3rs
8

Deﬁnition of “Animal”
by “Act on Welfare and Management of Animals”-“Animal” means any live animals
including both warm- and cold-blooded
animals (for instance, pets, agriculture
animals, wildlife animals and laboratory
animals)-

This term dose not exclude animals foruse in education and research, and for
agriculture.

-

This term excludes dead animals.
9

Demandof “Act on Welfare and Management of Animals”-

The “goal” of this law is to promote humanecare, use and treatment of animals.

-

To use animals for education and research,
the law directs the research community toaccept responsibility for the humane careand use of animals during all phases of theresearch eﬀort, according to the principlesof the Three Rs.

-

Researchers must euthanize animals, andthen put a terminate to any experiments
and operations.
10Penal provisions
by “Act on Welfare and Management of Animals”-

The Japanese government is trying tostop people from abuse animals by
amercement or imprisonment.

-

In the worst case, there is possibility
that Japanese court may be sentenced
to ﬁve yearʼs imprisonment or less.

11Standards Relating to the Care and Managementof Laboratory Animals and Relief of Pain
Notice No. 88 of the Ministry of the Environment, 2006

Care and management of laboratory animals
1) Fundamentals of care and management
2) Cage environment and animal room environment(1) Housing space(2) Environmental temperature and humidity(3) Ventilation (4) Lighting (5) Food (6) Water(7) Retention of Records

Health management of laboratory animals
Safety management and the other12Fundamental Guidelines for Proper Conductof Animal Experiment and Related Activitiesin Academic Research Institutions
Notice No. 71 of the Ministry of Education, Culture,
Sports, Science and Technology, 2007
Articles
1. Deﬁnitions
2. Responsibilities of the director of the academic research institution
3. Institutional Animal Experiment Committee
4. Conduct of animal experiments and related activities
5. Rearing and maintenance of laboratory animals
6. Miscellaneous Provisions

Inspection by the Institutional Animal Care and
Use Committee (IACUC)
Preparation of the animal use protocol
considering the principle of the 3Rs.
13

Self-regulation System to enhance animal
well-being for education and research use
To enhance well-being oflaboratory animals

To do animal experiments
properly

The Ministry of
the Environment

The other Ministries
cooperation

Propagating of the lawʼs principle

Administrative guidance

Philosophy

Regulation
Self-regulation system

Universities, and Research Institutes
The responsibility of the Institutional Oﬃcial and the role of the IACUC have been
determined by the oﬃcial documents.
The inspection of the IACUC and the other have been performed.
14

University Policy and Regulation,and the other
Policy and regulations of animal use and care, Okayama University- Notice No. 6 of President of Okayama University, 2008
http://www.cc.okayama-u.ac.jp/~animal/library̲main/iinkai/kisoku.pdf ←Japanese

Guide for the Care and Use of Laboratory Animals - Eighth Edition- by National Research Council of the National Academies, USA

Animal Research: Reporting of In Vivo Experiments (ARRIVE)
guidelines- by National Centre for the Replacement, Reﬁnement andReduction of Animals in Research (NC3Rs), Englandhttps://www.nc3rs.org.uk/arrive-guidelines

15

The role of the Animal Care and Use
Committee, Okayama University
The committee shall have the following duties;(i) Upon a request from the president of Okayama
University, the committee shall review the animal use
protocol submitted by the principal investigator todetermine whether it complies with the law, Care andManagement Standards, the Fundamental Guidelines
concerning animal experiments and related activities, andthe Institutional regulations,(ii) and shall report the results of the ﬁndings to the directorof the institution.
(iii)The committee receives a report from the director of theacademic research institution of the results of the animal
experiments performed according to the animal use
protocol,(iv)and shall give advice as required.
16

The role of the Animal Care and Use
Committee, Okayama University
President

consult

Executive Director for Research

reportapproval
Laboratories foranimal experiment

The Animal Use
and Care
Animal use protocol
Committee(IACUC)
issue
submitinspect

submitMembers:
Principal investigator

Managere.g. Dean / Director(Superviser)

Experimenter
Animal keeper

Representativeof the faculty,
Member of
Animal Resourcesincl. Students, Part­timer
17

What is needed to do research using
animals in Okayama University?
Attend the lecture
Approved laboratory for animal use
Approved animal use protocol…
Knowledge
Technique18

Where can we do animal experiments
in Okayama University?
Animal facilities
Facilities for care and keeping experimental
animals constitutively,
and for doing animal experiments
Laboratories for animal experiment
Laboratories for doing experimentswith live animals,
and for keeping experimental animals
temporarily ( 48 hrs)
19

Animal Facilities ofDepartment of Animal Resources (DAR)
Animal facilities in Shikata,
the core facility of DAR
Medical School
Dental School
and the others

Tsushima South (Tsuhima-minami)
Branch of DARʼs facilities
Faculty of Science,
Engineering, and Education

Faculty of Agriculture,and Pharmaceutical Sciences

Tsushima North (Tsushima-kita)
Branch of DARʼs facilities
20About locations (lab.) for animal use
outside animal facilities
In Okayama University, laboratories for animal experiments
need to be approved for animal use by the president,
after reviewing by the committee.
Each researcher must obey the following rules.
1) Using animals is not allowed in laboratories without approvalof the president.
2) Breeding and keeping (one month and over) of animals is notallowed in all the laboratory.
3) If the keeping of animal may run over 48 hours, the animal
use protocol, includes the description with rationale, must be
approved preliminarily.
4) Exhibit a label including bring in-date, species, strain, title, anduser name.
The animal use and care committee may conduct
inspections of your laboratories every other year.
21

Process of the protocol review
Principal investigator

1 wk
or moreDepartment of Animal Resources (DAR)
Shikata
Including conﬁrmation of
the compliance with Japanese laws

Check the form etc.
Pre-review by the committee members in DAR
2 wks
or moreThe committee members
Review

1 wk
or moreDAR Shikata
Notice of approvalor not by the president
Issue of certiﬁcations

Principal investigator
22

Animal Use Protocol for research and education
Animal Use Protocol is a detailed description of the
proposed use of laboratory animals.
In Okayama University, this is one of the most important
document about the animal use.
Consideration of the preparation of the protocol• Rationale and purpose of the proposed use of animals• Clear and concise sequential description of the procedure
involving the use of animals.
• Justiﬁcation of the species and number of animals proposed• Appropriate sedation, analgesia, and anesthesia• Appropriate conduction of the procedure of surgery, otherinvasion, and stress• Post-procedural care and observation• Description of rationale for anticipated or selected endpoint• Criteria and process for timely intervention, removal ofanimals from study or euthanasia if painful or stressful
outcomes are anticipated.
• Method of euthanasia or disposition of animals.
23

Animal Use Protocol for research and education

title
protocolpurposeimportance / signiﬁcancenecessity
person in charge

pain
categories
experimentersanesthesia
euthanasia

where to keep
and use animals

periodabout animals

approval by
the president

page 1

page 224

Frequently asked questions
about animal use protocol
Q Are there any sample forms?
A Yes, such as for breeding, freezing embryos, and frozen
A embryo transferring. Please ask to IACUC.
Q I want to do preliminary test.
A Please apply the animal use protocol just for
A preliminary test.
Q As I feel uneasy about the experimental technique, I
A want to practice before the actual test.
A Please apply the animal use protocol just for practice.

25

Choice of Anesthesia and Analgesia-> Select by technique / purpose
Widely accepted
anesthesia method
Barbiturates + analgesia
e.g. Secobarbital with Medetomidine

Features- Convenient- Large inﬂuence on the cardiovascular system- Better to use for perfusion ﬁxation or primary culture

Inhalant anesthetics
e.g. Isoﬂurane, Sevoﬂurane- Needs dedicated equipment (Inhalators are arrangedat each facility of DAR)

Ketamine combinations
e.g. with Xylazine or Medetomidine- The special license is needed to use ketamine- Wide margin of safety

Combinations with opioid
Butorphanol combinations with
Medetomidine and Midazolam- Troublesome adjustment- Stable induction of anesthesia- Long duration of anesthesia- Wide margin of safety

Low temperature anesthesia- Anesthetic eﬀect by lowering the temperature- Not available for large animals- Waterboarding is an abuse, so care must be taken toprevent animals from getting wet

26

Do not use as anesthesia
Drugs / Methods

Features- Easy- Narrow margin of safety

Barbiturates solely- Immobilized dose has no analgesic eﬀect- The committee does not recommend to use barbiturates
as an anesthetics solely for animal experiment

Diethyl ether- Respiratory irritation- Flammability- Regulated by law

Halothane- May cause health hazard- No analgesic eﬀect

Muscle relaxant solely

Urethane- Respiratory distress with no unconscious eﬀect- Cruel- Carcinogenic- Use with draft chamber
27

Acceptable euthanasia method
Drugs / Methods

Features

Overdose administrationof barbiturates- Simple- Pentobarbital was discontinued- Secobarbital 150mg/kg

Cervical dislocation
Decapitation- For experienced person- For mice and rats (Cervical dislocation:

CO2- Do not use dry ice- CO2 can be used in each ﬂoor of Shikata facility of DAR

Exsanguination- Do under anesthesia- For small animals (rodents, rabbits, etc.)

Intravenous injection of
KCl- Do under anesthesia- For large animals (pigs, dogs, etc.)- 20ml of KCl1mEq/ml for BW 30kg
- Use under anesthesia

Muscle relaxant- Poisonous drug
- Expensive

28

200g)

Experimental and Humane endpoints
Experimental endpoint of a study occurs when thescientiﬁc aims and objectives have been reached.

Humane endpoint is a point at which pain anddistress in an experimental animal is prevented, terminated,or relieved.

Special consideration
Tumor models, infectious diseases, vaccine challenge,
pin modelling, trauma, production of monoclonal antibodies,
assessment of toxicologic effects, organ or system failure,and model of cardiovascular shock.

Describe specific statement in the protocole.g. If the tumor diameter increases to XX cm or more,
and the condition of animals deteriorates,
the experiment will be stopped immediately andanimals will be euthanized. 29

What is needed to do research using
animals in Okayama University?
Attend the lecture
Approved laboratory for animal use
Approved animal use protocol…
Knowledge
Technique30

University Policy and Regulation,and the other
Policy and regulations of animal use and care, Okayama University- Notice No. 6 of President of Okayama University, 2008
http://www.cc.okayama-u.ac.jp/~animal/library̲main/iinkai/kisoku.pdf ←Japanese

Guide for the Care and Use of Laboratory Animals - Eighth Edition- by National Research Council of the National Academies, USA

Animal Research: Reporting of In Vivo Experiments (ARRIVE)
guidelines- by National Centre for the Replacement, Reﬁnement andReduction of Animals in Research (NC3Rs), Englandhttps://www.nc3rs.org.uk/arrive-guidelines

31

ARRIVE guidelines
Animal Research: Reporting of In Vivo Experiments

https://arriveguidelines.org/

32

National Centre for the Replacement
Reﬁnement & Reduction of Animals in Researchhttps://www.nc3rs.org.uk/
33

Practical training class ofmice and/or rats for beginner in DAR
Contents
Handling (including acclimatisation for handling)
Administration techniqueoral administration, intraperitoneal administration,and intravenous administration
Collecting bloodfrom tail vein, inferior vena cava, and heart
Euthanasia
Dissection

Class tuition
Held regularly in spring and autumn¥5,400 (mice) / ¥5,100 (rats)
Follow-up class¥2,300 (within 1hr)
Held temporary as you want (with 1hr follow-up class)¥7,500 (mice) / ¥7,200 (rats)
34

Videos of animal experiments technique35

How to apply to watch videos

１２36How to apply to watch videos

NameE-mail address

Agreement<- Submit!
37

For animal welfare
For animal welfare, as well as getting beneﬁt fromanimal experiments,
breeding, handling, methods, experiments, andrelated activities must be conducted with as little
pain and distress as possible to animals we can.
Letʼs attempt to communicate your supervisor
and/or Japanese colleagues before and/or
during animal use for research.

38

Letʼs start research!
You use live animals...
in your lab andthe animal facility

only in your lab

Attend this lecture and pass the exam on “Moodle”
Add your name to the listof the approved animal experiment protocol in Okayama Univ.
= ask your supervisor, PI, or professor,
and hand the application form to us
For Shikata
Please check our website

Attend the induction
for new usersof each animal facility

For Tsushima-north and -south
Please ask us!
39

Contact information
Laboratory for experiment animals, Shikata,
Department of Animal Resources,
Advanced Science Research Center, Okayama University
URL of the facility:
http://www.cc.okayama-u.ac.jp/~animal/
URL of the IACUC:
http://www.cc.okayama-u.ac.jp/~animal/committee.html
E-mail:iinkai@md.okayama-u.ac.jp
Extension number:
Shikata 7445

Do not forget to try and pass
the exam on Moodle!
1: Attendance conformation
2: Knowledge conformation40



Ministry of Education, Culture, Sports, Science and Technology, Notice No. 71
Fundamental Guidelines for Proper Conduct of Animal Experiment and Related
Activities in Academic Research Institutions under the jurisdiction of the Ministry of
Education, Culture, Sports, Science and Technology are provided as follows.
June 1, 2006
Kenji Kosaka 
Minister of Education, Culture, Sports, Science and Technology㻌 
Fundamental Guidelines for Proper Conduct of Animal Experiment and Related
Activities in Academic Research Institutions under the jurisdiction of the Ministry of
Education, Culture, Sports, Science and Technology PreambleA scientific understanding of the biological activities of living organisms on theplanet is essential for the solution of many issues, including human welfare, andconservation and restoration of the environment. Animal experiments and related
activities are necessary and unavoidable for gaining scientific insights into the
biological activities of living organisms, but they must be performed properly taking
into consideration the welfare of the animals. To ensure the appropriateness of animal experiments, efforts have been made inacademic research institutions to establish and implement institutional guidelines on
animal experimentation, through establishment of institutional animal experimentcommittees which conform to the Notice of “Animal Experimentation in Universities
and Similar Institutions.” (Director General, Science and International Affairs
Bureau, Ministry of Education, May 25, 1987). In the future, animal experiments andrelated activities are expected to continue, in order to facilitate advancement of thelife sciences and the development of medical treatments and other technologies.
Since then, the Law for Partial Amendment of the Law for the Humane Treatment
and Management of Animals (Law No. 68, 2005) was promulgated in June 2005. In
this Law, in addition to the provision of Refinement (i.e., techniques to reduce painand distress to animals used, while still allowing attainment of the scientificobjective) of the so-called 3R principles of animal experiment and related activities,
the provisions of Replacement (i.e., substituting animals with non-animal researchmethods, while still allowing attainment of the scientific objective) and Reduction(i.e., minimizing the number of animals used, while still allowing attainment of thescientific objective) were included.㻌 

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With these changes in the circumstances surrounding animal experiment andrelated activities, it is important for Academic Research Institutions to perform
properly animal experiment and related activities not just out of scientific necessity,
but also with consideration of both the scientific rationale and the welfare of theanimal, pursuant to the provisions of the Law for the Humane Treatment andManagement of Animals (Law No. 105, 1973; “the Law” hereinafter), as well asStandards Relating to the Care and Management of Laboratory Animals and Reliefof Pain (Notice No.88 of the Ministry of the Environment dated April 28, 2006;“Care and Management Standards” hereinafter).
On the basis of the current state of affairs, these Fundamental Guidelines for Proper
Conduct of Animal Experiment and Related Activities in Academic ResearchInstitutions (“the Fundamental Guidelines” hereinafter) are provided to ensure
proper conduct of animal experiments and related activities.

Article 1 Definitions
The following terms used in the Fundamental Guidelines are defined below:㻔㻝㻕Animal experiments and related activities



Utilization

testing,ofanimalsforeducation,

researchanddevelopment,

manufacture of biological products, or other scientific purposes.
(2)Laboratory animals

Animals of mammalian, avian or reptilian species, reared or maintained in
facilities of academic research institutions, for use in animal experiments andrelated activities.
(3)Academic Research InstitutionsAny of the following institutions where testing, research or development are
performed, in science, technology, or for academic research:
i Universities
ii Inter-University Research Institute Corporation
iii Colleges of Technology
iv Facilities and institutions under the jurisdiction of the Ministry of Education,
Culture, Sports, Science and Technology
v Independent Administrative Institutes (limited to those under the jurisdictionof the Ministry of Education, Culture, Sports, Science and Technology; excluding
the Independent Administrative Corporation Institute of National Colleges ofTechnology)
(vi) Corporations established pursuant to the provisions of Article 34 of the Civil

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

Code (Law No. 89 1896) (limited to those under the jurisdiction of the Ministry of
Education, Culture, Sports, Science and Technology)
(4) Animal experiment protocol
Protocol for conduct of animal experiment or related activities.
(5) Animal experiment researcher
Individual performing the animal experiment or related activities.
(6) Principal investigator
The animal experiment researcher who is responsible for all activities concerning
the animal experiment and related activities.

Article 2

Responsibilities of the director of the academic research institution

1. Responsibilities of the director of the academic research institution
The director of the academic research institution bears ultimate responsibility for
conduct of animal experiments and related activities in the institution. The director
shall establish the institutional animal experiment committee, establish theinstitutional regulations specified in Item 2 below, approve the animal experiment
protocols, comprehend the results of the animal experiments performed based on
the protocol, and implement any other necessary measures required for proper
conduct of animal experiments and related activities.

2. Establishment of institutional regulationsThe director of the academic research institution shall establish the institutional
regulations (“Institutional regulations” hereinafter) specifying methods for the
maintenance and management of animal experimentation facilities, as well as
methods for proper conduct of animal experiments and related activities, pursuant to
the provisions of the Law, Care and Management Standards, the Fundamental
Guidelines, and other laws and ordinances (including notices, the same hereinafter)
concerning animal experiments and related activities.

3. Approval of animal experiment protocolsThe director of the academic research institution shall have the principal
investigator submit the animal experiment protocol prior to the start of the animal
experiment and related activities, and shall approve or reject the submitted animal
experiment protocol after a review by the Institutional Animal ExperimentCommittee.


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4. Comprehension of the results of the animal experiment and related activities.
After completion of the experiment, the director of the academic research institution
shall receive the report of results of the experiment performed according to theanimal experiment protocol, and shall implement whatever measures are necessary
for improvement in the proper conduct of the animal experiment and related
activities, as required.

Article 3 Institutional Animal Experiment Committee
1. Establishment of the Institutional Animal Experiment Committee
The director of the academic research institution shall establish the Institutional
Animal Experiment Committee.
2. Roles of the Institutional Animal Experiment Committee The Institutional Animal Experiment Committee shall have the following duties.
(i) Upon a request from the director of the academic research institution, theCommittee shall review the animal experiment protocol submitted by the principal
investigator
Management

todetermineStandards,

whethertheitcomplies

Fundamental

with

the

Guidelines

Law,

Care

concerning

andanimalexperiment and related activities, and the Institutional regulations, and shallreport the results of the findings to the director of the institution.
(ii) The Committee receives a report from the director of the academic research
institution of the results of the animal experiments performed according to theanimal experiment protocol, and shall give advice as required.
3. Membership of the Institutional Animal Experiment Committee  The Institutional Animal Experiment Committee shall consist of members who areappointed from the following individuals by the director of the academic research
institution, with consideration given to achieve appropriately the roles of theCommittee.(i) Individuals with excellent knowledge and experience regarding animal
experiments and related activities.
(ii) Individuals with excellent knowledge and experience regarding laboratory
animals.
iii Individuals with scholarship and experience.

Article 4 Conduct of animal experiments and related activities
1 Assurance of scientific rationale  The principal investigator shall prepare the animal experiment protocol and shall

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properly perform the animal experiment and related activities, based on thefollowing items, from the standpoint of assuring the reliability of data obtained fromthe animal experiment and related activities.
(1) Selection of proper methods for the animal experiment and related activities
The animal experiment and related activities shall be performed by selecting theproper methods based on the following items.(i) Use of alternative methods (Replacement)
In the animal experiments and related activities, consideration must be given to
the proper use of laboratory animals by replacing laboratory animals with otherresearch materials where possible, while still allowing attainment of the scientificobjective.
(ii) Selection of laboratory animals (Reduction)
In the animal experiments and related activities, consideration must be given to
the appropriate use of laboratory animals by reducing the number of animals
used to as few as possible, while still allowing attainment of the scientificobjective. In these cases, it is necessary to consider the selection of laboratory
animal species appropriate for the purpose of the animal experiment and related
activities, the number of laboratory animals which will determine the accuracy
and reproducibility of the results of the animal experiment, the genetic andmicrobiological quality, and the rearing conditions.
iii Reduction of pain (Refinement)
In the animal experiments and related activities, methods must be employedwhich cause as little pain and distress as possible to the laboratory animals,
while still allowing attainment of the scientific objective, based on the Law andCare and Management Standards.
(2) Facilities and equipments for animal experiment and related activities
Animal experiments and related activities shall be conducted using appropriately
maintained and managed facilities and equipment.

2. Animal experiments and related activities that require special attention
concerning safety management  The director of the academic research institution shall consider the following
items when animal experiments and related activities are to be performed which
require special attention concerning safety management.
(i) When animal experiments and related activities are to be conducted which
involve physical or chemical agents or pathogens, or which may affect the safety or
health of humans or the surrounding environment, special attention to assuranceof safety and maintenance of the health of the animal experiment researchers
must be paid, while still properly maintaining facilities and equipment in the

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academic research institution.
(ii) The facilities and equipment shall be maintained so that the laboratory animals
are not harmed by contamination of the rearing environment, while consideration
shall also be given to maintenance of the health of the laboratory animals byimplementing quarantine or other measures as required.(iii) When animal experiment and related activities that might affect the ecosystem
are to be

performed, such as animal experiments or related activities using

genetically modified animals, special attention shall be given to prevention of
escape or other undesirable events of the genetically modified animals, while
facilities and equipment in the academic research institution shall be managed
properly.

Article 5 Rearing and maintenance of laboratory animals
Rearing and maintenance of laboratory animals in the animal experiment andrelated activities shall be performed properly from the standpoints of both scientificrationale and animal welfare, based on the Law and Care and Management
Standards.

Article 6 Miscellaneous Provisions
1. Conduct of education and training
The director of the academic research institution shall ensure that education andtraining are provided to animal experiment researchers and workers who engage inthe rearing and maintenance of laboratory animals (“animal experimentresearchers and workers” hereinafter), with the aim of guaranteeing that animal
experiment researchers and workers shall acquire the basic knowledge required forthe proper conduct of animal experiments and other activities, and for the proper
rearing and maintenance of laboratory animals. The director shall also implement
any other necessary measures to improve the level of skills and professionalism ofanimal experiment researchers and workers.

2. Self-inspections, evaluation and verification of compliance with the Fundamental
GuidelinesTo assure transparency concerning the conduct of animal experiments and related
activities, the director of the academic research institution shall periodically
implement self-inspections and evaluations of compliance with the Fundamental
Guidelines for the animal experiments and related activities in the institution.
Furthermore, the director shall endeavor to have the results of these self-inspections
and evaluations verified by a third party, independent of the research institution

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concerned.
3. Public disclosure of information
The director of the academic research institution shall publicly disclose
information regarding the animal experiments and related activities in theinstitution (e.g.,Institutional regulations, self-inspections and evaluations regarding
animal experiments and related activities, results of verification by a third party
independent of the institution concerned, and conditions of rearing andmaintenance of laboratory animals) through the Internet, distribution of annual
reports or by other appropriate means, approximately once a year. Supplementary Provision
The Fundamental Guidelines shall come into effect beginning June 1, 2006.

Reserch Promotion Bureau, Life Sciences Divisions -7-Guidelines for Proper Conduct of Animal Experiments
June 1, 2006
Science Council of Japan

Preface
The necessity of basic considerations for the handling of laboratory animals in Japan had been based on the
Law for the Humane Treatment and Management of Animals (Law No. 105, 1973) and Standards Relating to theCare and Management of Experimental Animals (Notice No. 6 of the Prime Minister’s Office 1980).
Under these conditions, the rationalization of animal experimentation was based on administrative
guidance rather than laws and regulations because of its importance in the advancement of scientific research. The
Science Council of Japan submitted a recommendation to the government in 1980 entitled “Establishment of Animal
Experimentation Guidelines.” In response to this recommendation, the Ministry of Education issued a notification
to related institutions entitled “Animal Experimentation in Universities, etc.” (Director General, Science andInternational Affairs Bureau, 1987). Based on this notification, research institutions established policies for more
appropriate conduct of animal experiments and Institutional Animal Care and Use Committees, and applied them indetail. As a result, it became possible to conduct highly creative scientific research in a free and open manner andJapanese medicine and life sciences made remarkable progress on an international level.
For progress in life science, it is recommended to have a voluntary system of animal experimentation under
the responsibility of researchers who best understand the necessity of such experimentation. There are also calls forthe exercise of government authority in animal experimentation. Therefore, establishment of guidelines on animal
experimentation became an urgent necessity and Subcommittee 7 of the Science Council of Japan issued a proposal
entitled “Promotion of public understanding of animal experimentation” in 2004.
On receipt of this proposal, the Ministry of Education, Culture, Sports, Science and Technology and
Ministry of Health, Labor and Welfare compiled “Fundamental guidelines for proper conduct of animal experimentand related activities in academic research institutions under the jurisdiction of the Ministry of Education, Culture,
Sports, Science and Technology” and “Basic policies for the conduct of animal experimentation in the Ministry of
Health, Labor and Welfare.” The two ministries requested the Science Council of Japan to prepare detailed
guidelines to serve as a reference material or a model when research institutions compile their own specifications foranimal experimentation in accordance with the above fundamental guidelines and basic policies.
Handling of laboratory animals is influenced by the religion and culture of each country. The so-called
North American model specifies voluntary management of animal experimentation without relying on legal
restrictions on scientific procedures, while Japan favors the establishment of a system based on Japanese customs.
With such a system, it is always hoped that animal experimentation will be promoted appropriately with theunderstanding of the people and will contribute to advances in life science research.
Index
Basis and objectives
No.1

Definitions
1) Animal experiment2) Facilities
3) Laboratory animal
4) Institution5) Director of institution6) Animal experiment protocol
7) Researcher(s)8) Principal investigator
9) Manager
10) Laboratory animal manager11) Animal technician
12) Manager, etc.
13) Policies
14) Regulations

No.2

Responsibility of the director of the institution

No. 3

Institutional Animal Care and Use Committee
1) Roles of the Institutional Animal Care and Use Committee12) Institutional Animal Care and Use Committee organization
No. 4

Animal experiment protocol drafting and experimental procedures
1 Drafting of animal experiment protocol
1) Items requiring consideration when drafting an animal experiment protocol
2) Animal experiment protocol form
2 Experimental procedures
1) Laboratory and laboratory equipment
2) Animal restraint
3) Food and drinking water restrictions4) Surgical procedures
5) Analgesic procedures, anesthetics and postoperative management
6) Humane endpoint
7) Euthanasia procedures
8) Safety management considerations
9) Reports of animal experiment results

No. 5

Laboratory animal selection and receipt
1) Introduction of laboratory animals
2) Quarantine and acclimatization
3) Transport
4) Provision of information on delivery and receipt of laboratory animals

No. 6

Care and management of laboratory animals
1) Fundamentals of care and management
2) Cage environment and animal room environment
 (1) Housing space (2) Environmental temperature and humidity (3) Ventilation
 (4) Lighting
 (5) Food (6) Water
3) Retention of records

No. 7

Laboratory animal health management

No. 8

Facilities

No. 9

Safety management
1) Understanding and dealing with risk factors
2) Prevention of injury by laboratory animals
3) Measures when laboratory animals escape4) Dealing with emergencies
5) Maintenance of the living environment

No. 10 Education and training
No. 11 Others
Additional provisions
Revision of these guidelines
References
Appendix2

Basis and objectives
Animal experiments are indispensable in medical and life science education, research and testing. Theyshould be managed and conducted voluntarily under the responsibility of each research institution. The researcher
must draft the animal experiment protocol based on scientific rationale and also should consider the welfare of theanimal. The researcher must have the Institutional Animal Care and Use Committee review the suitability of the
proposed animal experimentation protocol when conducting an animal experiment.
These Guidelines were prepared with the objective of appropriate implementation of animal experimentsfrom a scientific standpoint in accordance with policies on the conduct of animal experiments formulated bygovernment organizations with jurisdiction over institutions conducting animal experiments (Ministry of Education,
Culture, Sports, Science and Technology, Ministry of Health, Labor and Welfare, etc.) (“Fundamental guidelines forproper conduct of animal experiment and related activities in academic research institutions under the jurisdiction of
the Ministry of Education, Culture, Sports, Science and Technology” Notice of the Ministry of Education, Culture,
Sports, Science and Technology dated June 1, 2006 and “Basic policies for the conduct of animal experiments inresearch institutions under the jurisdiction of the Ministry of Health, Labor and Welfare,” Notification of the
Ministry of Health, Labor and Welfare dated June 1, 2006). The handling of laboratory animals is specified in“Standards Relating to the Care and Management of Laboratory Animals and Relief of Pain” (Notice No.88 of the
Ministry of Environment dated April 28, 2006).
These Guidelines consist of chapters on the responsibility of institutions concerning voluntary management
and the Institutional Animal Care and Use Committee at the beginning followed by chapters on animal experiment
protocol; drafting and experimental procedures and selection of laboratory animals. These are followed by care andmanagement of laboratory animals, laboratory animal health management, facilities and safety management,
education and training of personnel and others including in-house inspections and assessment and information
disclosure.
Each institution should formulate voluntary in-house regulations for proper scientific conduct of animal
experiments based on these Guidelines.
Note: These Guidelines are not intended for application in education, research or testing aimed at improvements incare and management of livestock or in breeding in the agricultural sector, but should be used in such fields as
required.

3No. 1

Definitions
In these Guidelines, the terms are defined as follows.

1) Animal experiment
Utilization of animals for education, testing, research, manufacture of biological products or other scientificpurposes
2) Facilities
Facilities and equipment used to perform animal experiments
3) Laboratory animal
Animal of mammalian, avian or reptilian species used in animal experiments
4) Institution
Organization (university, institute, independent administrative body, company, etc.) where animal
experiments are performed
5) Director of institution
Person with overall responsibility in the institution for proper and safe conduct of the animal experiments,(dean, director of an institution, principal of a school, chairperson of the board of directors, president, headof an institute, etc.)
6) Animal experiment protocol
Protocol drafted beforehand for the conduct of an animal experiment
7) Researcher(s)Person(s) performing the animal experiment8) Principal investigator
The researcher who is in charge of all duties related to the animal experiment protocol
9) Manager
Person in charge of the laboratory animals and facilities under the director of the institution (head of theanimal experimentation facilities, department head, etc.)
10) Laboratory animal manager
The laboratory animal manager assists the manager and is in charge of management of the laboratory
animals11) Animal technician
Person in charge of care and management of laboratory animals under the laboratory animal manager or
researcher12) Manager, etc.
Director of the institution, manager, laboratory animal manager, researchers and animal technicians
13) Policies
Fundamental guidelines and basic policies specified by government agencies related to animal experiments
and “Guidelines for Proper Conduct of Animal Experiments” (these Guidelines) specified by the Science
Council of Japan.
14) Regulations, etc.
In-house regulations of research institutions specified for the proper conduct of animal experiments and theproper care and management of laboratory animals based on related laws and ordinances and the policies

No. 2

Responsibilities of the director of the institution

The director of the institution bears the final responsibility for all experiments conducted in his or her
institution. The director of the institution prepares the facilities considered necessary for proper care andmanagement of the laboratory animals and proper and safe conduct of the animal experiments, appoints the manager
and appoints a person with knowledge and experience related to laboratory animals as the laboratory animal
manager. The director of the institution also provides education for related persons including the researchers andanimal technicians with the cooperation of the manager and laboratory animal manager to inform them of the related
laws and policies.
In each institution, in-house regulations including the authority and responsibilities of the director of theinstitution, standard operating procedures (SOP) for the conduct of animal experiments, proper care andmanagement of laboratory animals and methods of maintenance and management of facilities should be establishedbased on the policies.
An Institutional Animal Care and Use Committee should be established in each institution. The director of
the institution requests the Institutional Animal Care and Use Committee to review the animal experiment protocols
submitted by principal investigators based on scientific rationale and in consideration of animal welfare. The
director of the institution then approves or does not approve the protocol based on the report of the Institutional
Animal Care and Use Committee. After completion of the animal experiment, the director of the institution
examines the results obtained and instructs the principle investigator and manager to make improvements based on
advice of the Institutional Animal Care and Use Committee.

4The director of the institution retains the animal experiment protocols, results obtained from the animal
experiments and the minutes, etc. of the meetings of the Institutional Animal Care and Use Committee; assurestransparency of the animal experiments and publishes the results within a range that does not interfere with researchor corporate activities in consideration of protecting private information and research information. The director of
the institution should take the necessary measures to provide education and training to improve the quality oflaboratory animal managers, researchers and animal technicians.

No. 3

Institutional Animal Care and Use Committee

The Institutional Animal Care and Use Committee objectively reviews and inspects animal experiments at
an institution to assure that they are planned and conducted properly. To achieve this, the Institutional Animal Careand Use Committee should be established independently from any organizations involved in administration of the
facilities. The role and organization of an Institutional Animal Care and Use Committee are indicated below.
1) Roles of the Institutional Animal Care and Use Committee
Following consultation with the director of the institution, the Institutional Animal Care and Use
Committee reviews from the standpoint of scientific rationale the animal experiment protocol submitted by theprincipal investigator in consideration of the “Law for the Humane Treatment and Management of Animals”,“Standards Relating to the Care and Management of Laboratory Animals and Relief of Pain” and policies, andreports the results of the review to the director of the institution. The Institutional Animal Care and Use Committee
also receives the results of implementation of the animal experiment protocol from the director of the institution,and examines the actual conditions at the facilities, etc., as required before reporting back to the director of theinstitution and providing advice.
The Institutional Animal Care and Use Committee obtains details of the situation regarding the educationand training of laboratory animal managers, researcher(s) and animal technicians, and offers the director of theinstitution advice. The Institutional Animal Care and Use Committee may also participate in education and training
as required. Items discussed by the Institutional Animal Care and Use Committee are recorded as the meeting
minutes that must be maintained and retained. Institutional Animal Care and Use Committee meeting minutes
include the items below.
(1) Day, time and location of meeting(2) Names of members who participated in the meeting(3) Details of items discussed at the meeting (details of questions from committee members and answers
from principal investigators, etc.), and the results of discussions.
2) Institutional Animal Care and Use Committee organization
The Institutional Animal Care and Use Committee is composed of members appointed by the director of
the institution. To assure that committee members possess the knowledge required to fulfill the role of theInstitutional Animal Care and Use Committee, those appointed are researchers conducting animal experiments,
laboratory animal specialists and other persons of knowledge and experience.
The number of committee members is decided taking into consideration factors such as the size of theinstitution, the scope of the research, and the number of animal experiment protocols to be submitted. A committee
member should not participate in the review of an animal experiment protocol for an experiment for which he or she
is principal investigator.

No. 4

Animal experiment protocol drafting and experimental procedures

When conducting animal experiments, the significance of the research and the reasons why animal
experiments are required must be explained. Animal experiments must be conducted based on scientific rationale.
At the same time, they should be conducted in compliance with the internationally accepted 3R principles of animal
experimentation as clarified in amendments of the “Law for the Humane Treatment and Management of Animals”(Law No.105, 1973; the latest amendment on June 22, 2005), namely Replacement: the application of alternative
methods that do not require the use of animals within limits that allow scientific objectives to be achieved,
Reduction: the use of as few animals as possible within limits that allow scientific objectives to be achieved, andRefinement: the application of methods that do not distress the animals or subject them to pain within limits
required for use. These 3R principles are the ideology behind both animal experimentation and the handling oflaboratory animals. Consequently, within the limits required to achieve the objectives of research, they should betaken into consideration and applied appropriately when conducting animal experiments.
1.

Drafting of the animal experiment protocol
In accordance with the above principles, the principal investigator should prepare an animal experiment
protocol recording the necessary items in the form (2 below), and submit it to the director of the institution for5

approval. The director of the institution requests the Institutional Animal Care and Use Committee to review the
protocol content from a specialized standpoint. The Institutional Animal Care and Use Committee promptly reviews
the protocol and immediately reports the results of the review to the director of the institution. The animal
experiment can begin only after the principal investigator has received approval from the director of the institution.
The principal investigator conducts the animal experiment in compliance with the protocol approved by the
director of the institution. If changes to the protocol are required that go beyond the approved scope of theexperiment, procedures stipulated in the in-house regulations should be followed. After completion of theexperiment, a report to that effect should be submitted to the director of the institution in compliance with the inhouse regulations. If improvements indicated by the director of the institution are to be implemented, the principal
investigator should confer sufficiently with the laboratory animal manager as required.
Below are examples of items that the principal investigator should consider when preparing a protocol,
together with details of the animal experiment protocol form.
1) Items requiring consideration when drafting an animal experiment protocol* The objective and necessity of the animal experiment* Whether or not the animal experiment is unnecessary repetition* Whether an in vitro experiment could be conducted or the animal could be replaced by a phylogenetically lower
species (use of alternative methods)* Whether a change could be made to a less invasive animal experimentation method.
* The species of laboratory animals used and the genetic and microbiologic quality* The number of laboratory animals used
* Educational and training experience of the researcher(s) and animal technicians.
* Reasons why special cages and rearing environment are required* The anticipated disorders, symptoms and severity of pain resulting from experimental procedures* Measures to alleviate pain when it is anticipated that the laboratory animal will suffer severe pain
* The use of sedatives, analgesics and anesthetics* Whether major surgical procedures should be repeated* Postoperative management methods* Terminal treatment of laboratory animals (method of euthanasia, etc.)* Whether the animal experiment could possibly affect people or the environment. If so, required measures andprocedures* Issues concerning the occupational health and safety of the researcher(s) and animal technicians.
Concerning new animal experiments on as yet unrecognized research subjects, determining the experimental
method and number of animals to use may pose problems. In such cases, attempts should be made to prepare a final
protocol after conducting preliminary experiments to ascertain possible appropriate methods and number of animals.
For protocols that entail unavoidable, severe pain for the animals, the principal investigator should conduct literaturesearches to determine whether alternative methods are available. If there are no alternative methods, in cases where
the relief of pain through the use of measures such as anesthetics and analgesics is thought to be difficult, it isdesirable that advice be obtained from a laboratory animal specialist as required. When this is necessary, it should
be clearly noted in the protocol.
2) Animal experiment protocol form
It is advisable to prepare the animal experiment protocol form referring to the examples below.
(1) Principal investigator
Name
Affiliation and position
Contact address
Animal experiment experience and education and training(2) Name(s) of the researcher(s) [name(s) of subinvestigators](3) Research subject(4) Research objective(5) Detailed experimental procedure to be used on the laboratory animals(6) Term of animal experiment(7) Type of animal experiment (details of specific content)
Testing, research
Education and training
Other (enter specific details)(8) Laboratory animal species, strain, sex, age, etc, and number of animals used
(9) Location where the animal experiment be performed(10) Rearing methods (rearing location, group or individual rearing, number of animals per cage in the case ofgroup rearing, food)(11) Reasons why an animal experiment is necessary (specific details)

6

No alternative method
Sensitivity and precision of alternative methods are insufficient
Other (reasons: )(12) Severity of pain in animals to suffer due to each of the procedures (refer to pain classification indicated in
the Consensus Recommendation on Effective Institutional Animal Care and Use Committees drawn up by the
Scientists Center for Animal Welfare (SCAW), (Laboratory Animal Science. Special Issue: 11-13, 1987)(13) Pain alleviation methods for laboratory animals (specific details)
No measures implemented since pain is mild
No particular problems are thought to exist because restraint and constraint are short term
Anesthetics and analgesic will be used (drug name: )No pain alleviation method exists that does not prevent the scientific objective being achieved (reasons: )Long-term restraint and constraint are unavoidable (reasons: )A humane endpoint will be applied (endpoint determination: )Other ( )(14) Euthanasia procedures
Overdose of anesthetic
Inhalation of carbon dioxide
Cervical dislocation
Other ( )(15) Disposal of laboratory animal carcasses(16) Physical, chemical and biological risk factors, use of genetically engineered animals
2 Experimental procedures
When conducting animal experiments, pain suffered by the laboratory animals should be reduced as muchas possible within limits that allow the scientific objective to be achieved. Since scientific requirements differ foreach animal experiment, the principal investigator should describe specific experimental procedures and theanticipated severity of pain in the animal experiment protocol and receive approval from the director of theinstitution after review by the Institutional Animal Care and Use Committee.
The principal investigator should retain test reagents, drugs and laboratory equipment appropriately. In
particular, the laws and ordinances concerning the storage of controlled substances such as narcotics, poisons anddeleterious substances must be observed.
When conducting experimental procedures researchers should bear in mind the following points.
* Acquisition of skills in restraining laboratory animals, administering drugs, obtaining samples and othertechniques.
* Acquisition of skills relating to surgical procedures (prolonged operative procedures such laparotomy,
thoracotomy, craniotomy, orthopedic surgery and other procedures should be conducted under the guidance of a
specialist with sufficient knowledge and experience of those procedures.)* Pain relief procedures for laboratory animals.
* Observation of experiment discontinuation and completion criteria (humane endpoint).
* Acquisition of knowledge and skills related to euthanasia procedures.
1) Laboratory and laboratory equipment
Laboratories designed for performing experimental procedures on animals and also for analyzing
physiological functions should be constructed to prevent animals escaping and to enable easy cleaning anddisinfection to prevent contamination by excrement and blood. Clean, hygienic conditions should be maintained at
all times, and every effort made to organize the laboratory to assure that even if a laboratory animal escapes, it canbe easily recaptured.
Laboratories for surgical procedures (operating rooms) differ in equipment requirements depending on thephysical attributes of the study animal, the number of animals, complexity of the surgical procedure, the number andsize of devices used and other factors. Experiments using rodents can be conducted in an ordinary laboratory if it ispossible to use aseptic techniques to prevent microbial contamination in the surgical field. With large laboratory
animals, surgical procedures are often complex, surgery is prolonged and it is conducted by a surgical team. Thisrequires a correspondingly large operating table, inhalation anesthesia devices, surgical lighting, life monitoring
devices and other equipment. Support areas such as examination rooms, X-ray rooms and changing rooms in close
proximity to operating rooms should also be provided. In particular, when the objective is to conduct an animal
experiment in which the animal lives for a long period after surgery, preventive measures against contamination
should be implemented, the laboratory should be designed to enable easy cleaning after use and an air conditioning
system to supply clean air should also be taken into consideration.
When it is necessary to conduct experimental procedures on laboratory animals in an animal rearing room, theinfluence on other laboratory animals being reared in the same room should be minimized as much as possible. For
dogs, cats, monkeys and other animals exhibiting a high degree of emotional behavior, it is particularly important toassure that they do not feel any anxiety from sharing the same room.

72) Animal restraint
Physical restraint refers to localized or general restriction of the normal movements of laboratory animals
manually or with devices for examinations, sample collection, dosing and treatment. Restraining devices (restrainers,etc.) should be an appropriate size and easy to use, and should cause laboratory animals as little discomfort andinjury as possible. When using restraining devices, training of laboratory animals is required to enable them tobecome accustomed to the devices and researchers. With dogs, cats and monkeys, if they are conditioned for
aggressive restraint, their limbs extend outwards and they assume an immobile posture for short experimental
procedures in many cases.
Restraint for a prolonged period in a monkey chair or other device should be avoided unless it is essential forachieving the research objective. Light restraint such as a leash for restraining monkeys or other devices that do notinterfere with the natural posture of the animal is applicable within the range of experimental purposes. Items thatshould be considered concerning restraining devices are indicated below.
* A restraint period only as long as that required to achieve the research objective.
* Frequent observation of the condition of laboratory animals.
* Release from the restraining device of laboratory animals suffering from trauma or poor physical condition due
to restraint.
* Restraining devices should not be considered as rearing devices.
* Restraining devices should not be used as convenient tools for rearing management.
3) Food and drinking water restrictions
Achieving the objective of some research requires food and water restrictions for laboratory animals. Even if
it is indispensable to improve reliability and reproducibility of data from animal experiments, the following items
should be carefully considered.
* Even when food and water restrictions are required for experimental reasons, plans should be made so that the
animals ingest the minimum amount of food and water required.
* A scientific rationale is required for the restriction of food and water for research purposes.
* To monitor the state of dehydration, physiological and behavioral indices should be observed, and body weight
and other parameters measured.
4) Surgical procedures
When subjecting laboratory animals to invasive surgical procedures, particular attention should be paid to thefollowing points to alleviate pain as much as possible to within the limits that do not prevent the objective of theresearch being achieved.
* As well as aseptic techniques for the surgery itself, postoperative aseptic techniques and postoperative
management are important.
* Antibiotics should be administered in cases where the alimentary tract or other non-sterile site is surgically
exposed or where there is the possibility of reduced immune function due to the surgical procedure. However,administration of antibiotics is not an alternative for aseptic techniques.
* For major surgery (laparotomy, thoracotomy, craniotomy, etc.), aseptic techniques, anesthetic and analgesic
procedures, fluid infusion and keeping the animals warm are essential since body cavities are invaded andexposed resulting in substantial physical and physiological injury.
* For minor surgery (wound suturing, cannulation of a peripheral blood vessel, etc.), the conditions are not as strict
as those for major surgery since body cavities are not exposed and there is virtually no physical injury or it rarely
occurs. However, equipment must be sterilized and anesthetics should be used appropriately.
* Highly invasive, major surgery should be conducted under the guidance of a specialist with sufficient knowledge
and experience of the techniques involved.
5) Analgesic procedures, anesthetics and postoperative management
The alleviation of pain in laboratory animals is important not only from the standpoint of animal welfare but
also to assure the reliability and reproducibility of animal experiments.
* Analgesic procedures should be initiated when symptoms of pain are perceived in a laboratory animal. When an
animal feels pain, species-specific behavior includes vocalization, depressed behavior, abnormal expressions or
posture and lack of movement.
* To be able to perceive an abnormality, it is important to understand the behavioral, physiological andbiochemical characteristics of that species (or individual) when at rest and at ease.
* To select analgesic and anesthetic methods that do not interfere with the objective of the research, advice should
be obtained from a physician, veterinary surgeon, pharmacist or other specialist as required.
The degree of observation of animals required during the postoperative recovery period depends on the animal
species and the contents of the surgery. Attention should be focused on environmental temperature control,
monitoring of cardiovascular and respiratory function and postoperative pain, with particular attention paid tosymptoms of recovery from anesthesia.

8

* To deal with unexpected situations, advice should be obtained from a laboratory animal health management
specialist.
* Monitoring items include depth of anesthesia and physiological functions as well as evaluation of clinicalsymptoms and general condition.
* Maintaining normal body temperature is effective for preventing cardiovascular and respiratory disorders causedby anesthetics.
* During the recovery period after anesthesia, laboratory animals should be kept in a clean location at an
appropriate temperature and humidity, with their condition monitored frequently.
* Consideration should be given to parenteral infusions to maintain the water/ electrolyte balance, and toadministration of analgesics and other agents for management of the surgical field.
6) Humane endpoint
The humane endpoint refers to the timing of termination of an experiment (in other words, the timing of theapplication of euthanasia procedures) to release a laboratory animal from severe pain and suffering. It is a term usedin contrast to “death” as an endpoint that is used in protocols of animal experiments where the experiment continues
until the animal’s death.
* As a rule, euthanasia procedures should be available for termination of animal experiments.
* At the final stage of an animal experiment or when analgesics, sedatives or other agents do not provide relief,euthanasia procedures should be performed to release the laboratory animal from pain and suffering (one pain
relief method).
* Indications of when humane endpoint is applicable include food and water intake difficulties, moribund
symptoms (self-injurious behavior, abnormal posture, respiratory disorders, vocalization, etc.), abnormal
appearance over a prolonged period with no visible indications of recovery (diarrhea, bleeding, soiled genital area,etc.), weight loss (20% or more over several days), and marked increase in tumor size (10% or more of bodyweight).
* Reference should be made to pertinent international guidelines for details concerning determination of thehumane endpoint.
* When conducting animal experiments in which the degree of pain and suffering is high, such as lethal toxicity
studies, infection experiments and radiation experiments, the principal investigator should examine setting of thehumane point in the planning stage of the animal experiment.
7) Euthanasia procedures
When disposing of laboratory animals on completion of the experiment in accordance with the animal
experiment protocol or due to the laboratory animals being subjected to severe pain and suffering during the courseof the experiment when anesthetics and analgesics can not be used in the research, the researcher(s) should conducteuthanasia.
Selection of the agent and method used for the euthanasia procedure depends on the animal species and theobjective of the experiment. In general, a chemical method (overdose of a barbiturate anesthetic, administration of anon-explosive inhalation of anesthetic or carbon dioxide gas) or a physical method (cervical dislocation,
decapitation, exsanguination under anesthesia, etc.) is used. However, from the standpoint of animal welfare, theprincipal investigator should seek the advice and guidance of a laboratory animal specialist as required since there
are slight international differences on what are judged to be appropriate methods of euthanasia for laboratory
animals.
* Euthanasia procedures refer to procedures resulting in the rapid loss of consciousness and then death of alaboratory animal not associated with pain or suffering. In addition to Guidelines on Methods of Sacrificing
Animals (Notice No.40 of the Prime Minister's Office, July 4, 1995), international guidelines should be taken into
consideration.
* Euthanasia should be performed by methods that do not cause distress to other animals in the laboratory. Thisrequires careful attention because until animals lose consciousness they can vocalize and release pheromones.
* A person who has acquired the skills required for handling a particular animal species should conduct euthanasia
procedures, and the death of the animal should always be verified.
8) Safety management considerations
Genetic engineering experiments, animal experiments using radioactive materials or radiation, experiments
using poisons, deleterious substances or psychotropic drugs, and animal experiments using pathogenic agents or
hazardous chemicals must be conducted in strict compliance with related laws and ordinances. Animal carcasses andlaboratory waste must be disposed of appropriately using the methods stipulated in in-house regulations. In
particular, laws and ordinances related to waste material regulated by law must be respected.
9) Reports of animal experiment results
In compliance with in-house regulations after conducting an approved animal experiment, the principal
investigator must report the number of laboratory animals used, whether any changes were made to the protocol and9the results of the experiment to the director of the institution. The director of the institution should consult theInstitutional Animal Care and Use Committee on the appropriateness of the experiment as required.

No. 5

Laboratory animal selection and receipt

Microbiologic control of the rearing environment is particularly important, the same as genetic control oflaboratory animals. It can easily influence scientific reliability of animal experiments including accuracy andreproducibility of data. Consequently, when selecting animals for experiments, sufficient attention should be paid toboth genetic and microbiologic quality. Health management and safe rearing of introduced laboratory animals
should be based on “Standards Relating to the Care and Management of Laboratory Animals and Relief of Pain”
and the “Standards Relating to the Care and Management of Domestic Animals.”
1) Introduction of laboratory animals
The following points shall be taken into consideration when introducing laboratory animals.
* The animals must be procured legally. The laws relating to the delivery and receipt of genetically modified
animals and designated alien species, and the laws relating to the importation of rodents and primates must be
observed (Note).
* The use of purpose-bred laboratory animals with accompanying microbiologic monitoring records or infectious
disease test records is desirable. This animal production site data is useful for determining whether to accept thelaboratory animals or not.
* Each time a laboratory animal is introduced, the order requirements and abnormalities in external appearance
should be checked, and quarantine and acclimatization performed according to the animal species and conditionsof the facilities.
Note: To prevent the occurrence of infectious diseases in humans due to animals when importing living mammals
and birds and the carcasses of rodents and lagomorphs, the import of animals is controlled by the “Law Concerning
the Prevention of Infectious Diseases and Medical Care for Patients with Infectious Diseases,” the “Enforcement
Regulations of the Law Concerning the Prevention of Infectious Diseases and Medical Care for Patients ofInfectious Diseases” and other regulations. The system of notification of import also applies to rodents to be used aslaboratory animals. In addition, when introducing laboratory animals that fall under the category of invasive alien
species (cynomolgus monkeys, rhesus monkeys, Taiwan monkeys, etc.), procedures must be completed incompliance with the “Invasive Alien Species Law” and related laws; for laboratory animals that fall under the
category of designated animals (Japanese macaques and other monkeys, poisonous snakes, etc.), the “Standards for
Designated Animals” based on the “Law for the Humane Treatment and Management of Animals” apply; forlaboratory animals that fall under the category of genetically modified animals, the “Law Concerning theConservation and Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified
Organisms” and related laws apply; for laboratory animals that fall under the category of domestic animals (pigs,
sheep, goats, etc.), the “Domestic Animal Infectious Disease Control Law” and related laws apply; for dogs, theRabies Prevention Law and related laws apply.
2) Quarantine and acclimatization
To prevent the spread of infectious diseases in facilities, newly introduced laboratory animals should be
kept away from animals already present until their health condition has been verified. If symptoms are observed orwhen deemed necessary, microbiologic examinations should be performed. The following items regarding
quarantine and acclimatization must be taken into consideration.
* Depending on the construction and sanitary conditions of the facilities, animal species and the objective of theanimal experiment, the manager should respect the opinions of the laboratory animal manager andcomprehensively determine the infectious diseases to be eliminated in the facilities.
* The principal investigator and laboratory animal manager should discuss necessary microbiologic control for
individual animal experiments.
* Reports of microbiologic monitoring at the supply source can be used as quarantine reference materials.
Obtaining information from the producer is important when considering quarantine procedures.
* The laboratory animal manager should determine the required quarantine period, risk to personnel and otheranimals, and whether treatment is required during the quarantine period.
* For mice, whether or not microbiologic cleaning by in vitro fertilization, embryo transfer or Caesarian section isrequired should be determined.
* Before use in animal experiments, it is necessary to set up an acclimatization period from the physiological,
psychological and nutritional viewpoints for the laboratory animal. The acclimatization period differs dependingon the method of transport and the required period of time, the animal species and intended purpose of thelaboratory animal used.
* Primates should be given sufficient time to become acclimatized to the environment with respect to both
personnel and laboratory equipment.

103) Transport
Transporting laboratory animals refers to moving laboratory animals between facilities. The following
items should be taken into consideration during transport.
* Personnel involved in the transport of laboratory animals should endeavor to take care of the health and safety of
the laboratory animals and to prevent any hazard, etc., for humans occurring due to the animals.
* In addition to causing laboratory animals fatigue and distress, transport influences data from animal experiments.
To conduct scientifically correct animal experiments, transport should be completed in as short a time as possible.
* During transport, laboratory animals should be supplied with food and water as required while maintaining anappropriate temperature using air conditioning, ventilation, etc.
* To prevent environmental contamination by laboratory animals during transportation, care and management
should be undertaken in essentially the same way as at the institution. Since transport involves the use of public
transportation or highways, every effort should be made to prevent environmental contamination in the event of
unforeseen circumstances. To do this, containers that not only prevent laboratory animals from escaping, but also
from which it is difficult for microorganisms, waste material, etc., to reach the exterior should be used.
* Containers used for transport should have the structure and strength to prevent laboratory animals escaping. They
should also be lightweight and compact, designed so they do not easily fall over, have lids that do not open due tovibrations, etc., and be well ventilated. In case laboratory animals show abnormal symptoms, escape, etc.,
containers should display a contact address (address, telephone number, etc.).
* When transporting across borders, the “International Air Transportation Association (IATA) Live Animal
Regulations” concerning the transportation of live laboratory animals require careful consideration.
4) Provision of information on delivery and receipt of laboratory animals
Persons who supply or sell laboratory animals should provide information and explanations concerning theanimals’ physiology, ecology, behavior and the correct care and management methods for the animals,
microbiologic quality, infectious diseases, etc. At facilities receiving animals, in addition to quarantine, appropriate
microbiologic cleaning when required (in vitro fertilization, embryo transfer, Caesarian section, foster nursing, etc.)
or administration of drugs, vaccinations should be provided as required.
On delivery and receipt of genetically modified laboratory animals, the “Law Concerning the Conservationand Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified Organisms,”“Ministerial Ordinance providing Containment Measures to be taken in Type 2 Use of Living Modified Organisms
for Research and Development” and other regulations must be observed.

No. 6

Care and management of laboratory animals

At facilities, laboratory animals must be correctly cared for and managed to increase the scientificreliability of animal experiments in consideration of animal welfare while also assuring the safety of theresearcher(s) and animal technicians. In this respect, the following items should be taken into consideration.
* The objective of care and management (testing, research, education, etc.).
* Individual characteristics such as species, strain, sex, age, physical attributes, behavior, history and healthcondition.
* Whether authorization is required for care based on related laws (when caring for and managing animals that fallunder the category of designated animals according to the Law for the Humane Treatment and Management of
Animals or invasive alien species according to the Invasive Alien Species Law, authorization is required from thegovernment or local authorities).
* Individual or group housing* Rearing period* Details of procedures in animal experiments (physical invasiveness, degree of invasiveness, items for daily
observation, etc.)* Genetic engineering procedures, immunosuppression procedures, infectious agents or administration ofcarcinogens.
1) Fundamentals of care and management
Laboratory animal managers and animal technicians should care for and manage laboratory animals withthe aim of maintaining the inherent physiology, ecology and behavior of the animals and minimizing stress as muchas possible. The manager, etc. should independently decide on the ideal rearing environment for laboratory animals
taking into consideration the scientific objectives. When caring for and managing different animal species or
numerous laboratory animals in the same facilities, the animals should be accommodated taking into consideration
this combination within the limits to assure it does not prevent the objective of animal experiments, etc., being
achieved. Animal technicians should supply the animals with food and water appropriately within the limits toassure that it does not interfere with the objective of animal experiments, etc., being achieved. In the event of11

facilities, etc., being closed, transfer to other facilities of the animals present should be considered for the effective
use of laboratory animals.
2) Cage environment and animal room environment
The environments inside cages and in the animal room are connected by ventilation but differences intemperature, humidity, etc., may occur. In general the temperature, humidity and concentration of gaseous andparticulate substances reach higher levels inside cages than in the animal room. If attention is not paid to this point,
it may unexpectedly influence the metabolism, physiological function, etc., of the laboratory animals being reared,or alter their susceptibility to disease.
The following items concerning cages and other rearing equipment should be taken into consideration.
* Cages with the structure and strength to prevent escape of the animals* Easy access to food and water for individual laboratory animals* Maintenance of normal body temperature* Urination, defecation and preservation of natural posture* Depending on the behavior inherent to the animal species, the cage and laboratory animals must be kept clean
and dry.
* Depending on typical behavior of the animal species, social contact between animals and the formation ofhierarchies* Cages safe for laboratory animals (no sharp edges or protrusions, no gaps in which an animal’s body or limbs canbecome wedged)* As far as possible, observation without interfering with the animal behavior* Easy supply of food and water and easy changing of feeding and watering devices* Cage structure permitting easy cleaning, disinfection, sterilization etc., and cage materials able to withstand theseprocedures* Necessity and frequency of replacement of bedding, etc.
(1) Housing space
Since determining whether housing space is appropriate involves various factors, only taking into
consideration an animal’s weight and cage size alone is not sufficient. Rather than simply a larger floor area,
increased cage height or increased cage wall area, a sheltered area, or complex cage structures are required for someanimal species. Whether housing space is appropriate can be determined taking animal traits and behavior as indices.
In addition to referring to literature references such as the “ILAR Guide for the Care and Use of Laboratory
Animals”, the opinions of specialists and the necessity of conducting the research should be taken into consideration.
(2) Environmental temperature and humidity
To assure that homeothermal animals are comfortable, their body temperature should be maintained within
a normal range. Scientifically based recommendations for the appropriate range of temperature and humidity
required by different animal species are specified in textbooks and other sources. For laboratory animals notcompletely awake after being anesthetized for a surgical procedure, laboratory animals without hair, newborn
animals separated from their mother, chicks within a few days after hatching, the room temperature setting shouldbe raised or topical warming is required.
(3) Ventilation
The objective of ventilation is to supply the proper degree of oxygen by creating a static pressure difference
between adjacent spaces. Ventilation also eliminates the heat load generated by animals, lighting equipment andother devices and equipment, reduces the concentration of gaseous and particulate substances, and regulates room
air temperature and humidity. Ventilation inside cages does not necessarily reflect ventilation in the animal room.
An air conditioning system is extremely effective to maintain the proper laboratory animal rearing environment.
Consequently, as well as understanding the operating condition of the air conditioning equipment, measurement ofanimal room temperature, humidity, air change rate is required together with periodic inspection and maintenance ofequipment.
(4) Lighting
Lighting has physiological and morphological effects on various animals and can alter behavior.
Inappropriate lighting periods, lighting intensity and light spectrum cause laboratory animals stress. The fact thatmany laboratory animals frequently used are nocturnal must be taken into consideration. In addition, it should be
remembered that albino rats are more susceptible to retinopathy due to phototoxicity than non-albino animals, andeven a small amount of light entering the room during the dark period when the lights are off can influence the
estrus cycle of rodents.
(5) Food
Unless instructed otherwise by the principal investigator, the animal technician should correctly provide
laboratory animals with suitable nutritional, uncontaminated food every day. Attention should be paid to pathogens,vectors that transmit them (insects, etc.) and chemical contaminants that may be in the food so that they are notintroduced into the facilities. The area where food or its ingredients are processed and stored should be kept clean
and measures to prevent infestation by insects should be taken. Based on the storage temperature, food quality andother factors, the storage life of each kind of food should be determined by the laboratory animal manager. Feeding12devices that enable easy food intake should be used and care is required to assure they do not become contaminatedwith feces and urine.
(6) Water
The animal technician should check water bottles and automatic watering systems every day to verify thatthey are functioning correctly and kept clean. Animals unaccustomed to automatic watering systems may fall into astate of dehydration if observation is neglected. In such cases, the animals should be carefully trained to use thesystem. To prevent the transmission of microorganisms, rather than refilling water bottles it is preferable that thebottles themselves be changed.
3) Retention of records
The manager, etc. should keep record books and ledgers related to laboratory animal sources, rearing
history, history of disease, etc., and strive to manage laboratory animal records properly to aid in laboratory animalcare and management in facilities. This kind of documentation is also useful for assessing data reliability of animal
experiments.

No. 7

Laboratory animal health management

Laboratory animal health management should be conducted scientifically. To prevent laboratory animals
suffering injuries unrelated to the objective of the animal experiment or contracting a disease, the laboratory animal
manager and researcher(s) should conduct necessary health management. In the event of injury or disease unrelated
to the objective of the animal experiment, appropriate treatment should be provided within the limits that do notinterfere with achieving the objective of the animal experiment. For this purpose, there should be exchange of
information related to laboratory animal health conditions between the laboratory animal manager, the researcher(s)and the animal technician, and necessary measures should be implemented as quickly as possible.
Since laboratory animal health management requires understanding of animal species ecology, behavior
and physiological and anatomical characteristics, the ability to distinguish when they are normal or abnormal andthe ability to distinguish whether abnormalities are due to experimental procedures or other factors, the cooperationof all those involved as well as the laboratory animal manager is essential. If required, the advice of specialists on
individual animal species, diseases, etc. should be sought.
When conducting laboratory animal health management, preventing infectious diseases influencing animals
and humans as well as the experimental results is particularly important. Depending on the animal species and theobjective of the animal experiment, the need to conduct quarantine, isolation and microbiologic monitoring must be
conducted. When examining these points, the following items must be into consideration.
* Reported cases of infection of humans by laboratory animals include lymphocytic choriomeningitis originatingfrom mice and hamsters, hemorrhagic fever with renal syndrome from rats and herpes B virus infection andbacillary dysentery from primates.
* When using bird species in animal experiments, infections by Chlamydia psittaci and salmonella require special
attention.
* With regard to turtles, salmonella infection from green turtles has been reported.
* When rearing imported primates, advanced notification of the facilities and notification if there is an outbreak ofbacillary dysentery are required in compliance with the regulations stipulated in the “Law Concerning thePrevention of Infectious Diseases and Medical Care for Patients of Infectious Diseases” and related laws.
* When conducting laboratory animal health management using domestic animals, consideration must be given to
infectious diseases (domestic animal infectious diseases, notifiable infectious diseases) pursuant to the provisionsof the “Domestic Animal Infectious Disease Control Law” and related laws and ordinances, and diagnosis andadvice from a veterinarian must be sought if any abnormalities are observed.
* One important issue concerning quarantine is the detection of inapparent infections. Prevention of the
provocation of an inapparent infection due to stress caused by experimental procedures assures the reliability ofexperimental results as well as preventing the spread of infections diseases in facilities. Items to be consideredregarding quarantine are detailed in part 5.
* With some animal species, even if the pathogenicity of the pathogen is low resulting in an inapparent infection,
there are cases where other animal species become infected and develop a disease, so isolated rearing should be
provided for each animal species as a general rule.
* After an animal experiment has started, in addition to early detection and treatment of abnormalities byobservation of laboratory animal symptoms, periodic microbiologic monitoring is effective for verifying healthconditions in small animals such as mice, rats, guinea pigs and rabbits. The results of microbiologic monitoring
are also useful for preventing infectious diseases in facilities.
* When providing another institution with laboratory animals, the provision of a health certificate issued by aveterinarian may be required. When facilities cannot conduct their own examinations or issue a health certificate,a laboratory animal checking organization can be requested to do so.

13

No. 8

Facilities

Respecting the opinions of the laboratory animal manager, the manager should provide the conditions
necessary to meet the requirements for conduct of the research, for animal physiology, ecology and behavior, andsanitary management while establishing and administrating the facilities. Cleaning and disinfecting floors, insidewalls, ceilings, auxiliary equipment in the facilities should be easy, and the structure should facilitate maintenanceof sanitary conditions to eliminate the possibility of laboratory animals being injured by projections, holes,
depressions, inclined planes, etc. Inspections and maintenance of cages should be conducted to prevent laboratory
animals escaping, to keep laboratory animals physically comfortable, and to make hygiene management and daily
operations easy. The following items should be considered when setting up facilities.
* Equipment for care and management of laboratory animals, sanitation equipment for cleaning and disinfecting
materials and devices, and experimentation equipment* Assurance of structures and strength to prevent entry of wild animals* Assurance of structures and strength to prevent laboratory animals escaping* In facilities where animal experiments on infection with pathogens or using radioactive materials are conducted,
effective equipment to prevent escape of infected animals, laboratory animals administered chemicals andlaboratory animals treated with radioactive materials* Facilities with the structure necessary to reduce odor and noise and storage facilities required for waste material.
* Within the limits that do not interfere with achieving the objective of the animal experiment, proper space,temperature, humidity, ventilation, lighting, etc., to assure laboratory animals are not subjected to excessive stress* Depending on the animal species and the objective of the experiment, air conditioning equipment necessary tomaintain a constant environment in the facilities* In facilities for rearing laboratory animals extremely susceptible to infectious diseases, sanitation and air
conditioning equipment necessary to control microorganisms* Assurance of structures that enable researcher(s) and animal technicians to work without risk.
* Facilities and equipment such as safety cabinets, draft chambers and localized air exhaust equipment to prevent
work related accidents as required. Also education and training of researcher(s) and animal technicians tofamiliarize them with safe operating procedures.
* When conducting gas sterilization of rearing equipment such as vinyl isolators, gas masks should be worn as
required.
* For autoclaves and ethylene oxide gas sterilizers, periodic inspections as indicated by laws and ordinances, aswell as daily checks before starting work

No. 9

Safety management

The director of the institution should endeavor to assure health and safety during operation of the facilities
based on related laws and ordinances. Necessary facilities should be provided and health management to prevent
laboratory animal managers, researcher(s) and animal technicians from contracting diseases from laboratory animals
should also be conducted. For animal experiments that involve materials posing a physical or chemical risk or thatinvolve pathogens (“risk factors” hereinafter), and animal experiments using living modified organisms, it must be
assured that they do not have any influence on human or laboratory animal safety and health or the ecosystem, andthat disturbance of laboratory animals does not reduce the reliability of experimental results. Measures required toprevent disruption of public health, living environment and the ecosystem must also be implemented.
1) Understanding and dealing with risk factors
The director of the institution and the manager should have an understanding of the risk factors related tooccupational safety and health. When assessing risk factors, the opinion of a specialist should be sought if necessary.
At the planning stage of an experiment, the principal investigator should explain to the laboratory animal manager
the types of risk factors and degree of risk involved, and request cooperation in care and management locations andthe use of safety equipment for laboratory animals with risk factors. The principal investigator should also provide
the researcher(s) and animal technicians with necessary information concerning risk factors, the degree of risk,
methods of dealing with risk factors and accident prevention.
* To assess the degrees of risk posed by pathogens and chemical agents, reference should be made to related
guidelines and databases.
* In areas or rooms where risk factors are present, the risk factors should be displayed. For genetic engineeringexperiments and animal experiments that use ionizing radiation, labeling is required as specified by law.
2) Prevention of injury by laboratory animals
Typical risk factors concerning animal experiments include infection of humans by a laboratory animal
naturally infected with a pathogen, allergies caused by laboratory animal hair, etc., and bites and scratches bylaboratory animals. The laboratory animal manager, researcher(s) and animal technicians should exchange
information concerning the handling of animals posing such risks to prevent injury.

14

* To prevent being bitten by a laboratory animal when conducting an animal experiment, researcher(s) and animal
technicians should receive prior education and training.
* Emergency medical supplies should be available and a system enabling prompt emergency measures by aphysician at the time of accidents involving bites by poisonous animals such as poisonous snakes, as well as
primates, dogs should be established.
* Measures required to prevent people not involved in the care and management of laboratory animals or in the
animal experiment from touching laboratory animals should be taken.
3) Measures when laboratory animals escapeThe manager, etc., should implement measures necessary to prevent laboratory animals escaping from afacility where they are being kept. Outside of working hours, cage lids should be properly closed and cage doors
should be locked.
* The animal room door should be closed at all times and locked when necessary.
* The number of laboratory animals should be checked when starting and when finishing work.
* In-house regulations as a safeguard in anticipation of a laboratory animal escaping from the facility should beestablished* If a laboratory animal that might injure or cause harm to humans escapes from a facility, the relevant authorities
should be contacted at once.
4) Dealing with emergencies
In coordination with the government agencies concerned, the manager should take measures to comply
with anti-disaster measures, and draw up plans beforehand related to measures to be implemented in the event of an
emergency such as an earthquake or fire.
* In an emergency situation, measures should be taken to protect laboratory animals and prevent injury or harm tohumans or problems related to environmental protection arising due to escape of laboratory animals.
* A network for communication on holidays, at night and at times of emergency should be established, together
with an emergency communication system.
5) Maintenance of the living environment
The manager, etc. should treat laboratory animal waste material appropriately, keep the facilities, clean at
all times, and prevent contamination of the environment by microorganisms, spread of odors, and infestation by
insects. Also noise-proofing should be provided.
* Carcasses of laboratory animals and laboratory waste must be treated appropriately in compliance with wastematerial classification of the local authorities concerned. When carcasses and animal waste are temporarily stored,measures to prevent spread of odors and entry of insects and to assure sanitation should be taken.
* Syringes and needles used in animal experiments should be treated as infectious medical waste, collected inbiohazardous material containers, stored carefully to prevent the contents of the containers from escaping, anddisposed of in accordance with the regulations of the local authorities concerned.

No. 10 Education and training
The director of the institution should endeavor to provide separate education and training of the laboratory
animal manager, researcher(s) and animal technicians as required. Education and training should be provided prior
to engaging in animal experiments and should also be provided later as required.
Education and training should be conducted in accordance with in-house regulations, and the dates of
instruction, educational content and the names of the instructor and those receiving instruction should be recorded
and retained.
Education and training content should be specified in the in-house regulations taking into account activities
undertaken in the institution. From the viewpoint of proper conduct of animal experiments, the following items
should be included in education and training.
* Items related to pertinent laws and ordinances, bylaws, guidelines, in-house regulations* Items related to animal experiments, etc., and the handling of laboratory animals* Items related to the care and management of laboratory animals* Items related to safety assurance* Items related to the use of facilities

No. 11 Others
With the aim of implementing scientific animal experiments, taking into consideration animal welfare, thedirector of the institution should conduct periodic in-house inspections and assessments to determine whether theexperiments comply with policies and in-house regulations. Consideration should be given to receiving verification15

by a person or persons not connected with the institution. Records of in-house inspections and assessment records
should be retained for a fixed period of time.
Information related to animal experiments conducted at the institution based on in-house regulations should
be disclosed by the director of the institution by a method determined appropriate in accordance with effects on theprotection of personal and research information and proper corporate activities. Efforts should be made to improve
the social transparency of information related animal experiments in the institution.

Additional provisions
Revision of these guidelines
These guidelines should be revised in accordance with developments in related areas of research andchanges in society, as well as amendments of related laws and ordinances as required.

16

References
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3)
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Related laws and ordinances, policies, etc.
Law for the Humane Treatment and Management of Animals
http://www.env.go.jp/nature/dobutsu/aigo/amend_law2/index.html
Standards relating to the Care and Management of Laboratory Animals and Relief of Pain
http://www.env.go.jp/nature/dobutsu/aigo/anim_guide/index.html
Guidelines on Methods of Sacrificing Laboratory Animals
http://www.env.go.jp/nature/dobutsu/aigo/anim_guide/index.html
Ministry of Education, Culture, Sports, Science and Technology: Fundamental
guidelines for Proper Conduct of Animal Experiment in Academic Research Institutions
Ministry of Health, Labor and Welfare: Basic Policies for the Conduct of Animal Experimentation
http://www.mhlw.go.jp/general/seido/kousei/i-kenkyu/index.html㻌
Law concerning the Conservation and Sustained Use of Biological Diversity through Regulations on the
Use of Living Modified Organisms
http://www.maff.go.jp/carta/15hou97.pdf
Invasive Alien Species Law
http://www.env.go.jp/nature/intro/
Law concerning Prevention of Infectious Diseases and Medical Care for Patients with Infectious Diseases    
http://wwwhourei.mhlw.go.jp/cgi-bin/t_docframe.cgi?MODE=hourei&DMODE=
CONTENTS&SMODE=NORMAL&KEYWORD=&EFSNO=361
Domestic Animal Infectious Disease Control Law 
Rabies Prevention Law
http://wwwhourei.mhlw.go.jp/cgi-bin/t_docframe.cgi?MODE=hourei&DMODE=㻌CONTENTS&SMODE=NORMAL&KEYWORD=&EFSNO=403
Director General, Science and International Affairs Bureau, Ministry of Education, Culture, Sports,
Science and Technology : Animal Experimentation in Universities, etc. (Notification) 1987
Research Promotion Bureau, Ministry of Education, Culture, Sports, Science and Technology: Introduction of Laboratory Animals in Universities, etc.㸦Notification㸧2001
Science Council of Japan: Establishment of Animal Experimentation Guidelines㸦Recommendation㸧
1980. To refer to the data, contact the Science Council of Japan㸦03-3403-1091).
Science Council of Japan: Report of the Special Committee on a Social Consensus on Promotion of Life
Science Handling of Animals in Education and Research, 1997
To refer to the data, contact the Science Council of Japan㸦03-3403-1091).
Report of Subcommittee 7, Science Council of Japan Promotion of public understanding of animal
experimentation 㸦Proposal㸧2004
http://www.scj.go.jp/ja/info/kohyo/pdf/kohyo-19-t1015.pdf
Science Council, Ministry of Education, Culture, Sports, Science and Technology, Special Research
Field Promotion Committee, Biosciences, Subcommittee: Manual of Safety management ofMicroorganisms for Research in Universities, etc. (Draft) 1998

2.

Commentaries on related laws and ordinances and textbooks, etc.
Ten commentaries and textbooks written in Japanese (omitted)

3.
1)International policies and textbooks
Russell W & Burch R. The Principles of Humane Experimental Technique. Chapter 4. The source,
incidence, and removal of inhumanity. The removal of inhumanity: The 3R’s. 1957.
(http://altweb.jhsph.edu)
ICLAS (International Council for Laboratory Animals). Guidelines for the regulation of animal
experimentation. 1974.
CIOMS involving animals. 1985. (Council for International Organizations of Medical Sciences)/WHO.
International guiding principles for biomedical research
ILAR (Institute for Laboratory Animal Research)-NRC (National Research Council). Guide for the careand use of laboratory animals (7th edition). National Academy Press. 1996㸬
European Communities (currently European Union). Council Directive 86/609/EEC. 1986.
SCAW (Scientists Center for Animal Welfare). Categories of biomedical experiments based on increasing
ethical concerns for non-human species. 1987.

2)
3)
4)
5)
6)

17

7)
8)
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12)

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CCAC (Canadian Council on Animal Care). Guidelines on choosing an appropriate endpoint in experiments
using animals for research, teaching and testing. 1997.
ILAR-NRC. Humane endpoints for animals used in biomedical research and testing. ILAR Journal 41 (2)Special edition. 2000.
UK Coordinating Committee on Cancer Research. UKCCCR guidelines for the welfare of animals inexperimental neoplasia. 1988.
AVMA (American Veterinary Medical Association). 2000 Report of the AVMA panel on euthanasia.
JAVMA 218 (5), 669-696.
Working Party of the European Commission. Recommendation for euthanasia of experimental animals Part
1. Laboratory Animals 30, 293-316, 1996 & Part 2. Laboratory Animals 31,1-32, 1997
EFPIA (European Federation of Pharmaceutical Industries Associations) & ECVAM (European Centre for
the Validation of Alternative Methods). A good practice guide to the administration of substances andremoval of blood, including routes and volumes. 1999.
Frecknell P. Laboratory animal anesthesia (2nd ed). Academic Press. 1996.
Smith & Swindle (eds.). Research animal anesthesia and surgery. SCAW. 1994.
IATA (International Air Transportation Association). IATA live animal regulations. 1995 (22nd edition).
U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, and National
Institutions of Health. Biosafety in microbiological and biomedical laboratories (BMBL) (4th ed.)1999.
U.S. National Research Council, Occupational Health and Safety in the Care and Use of Research
Animals, 1997
Hau J & van Hoosier, Jr. G. Handbook of laboratory animal science (2nd ed.). Vol. 1 Essential principles

Appendix
Members of No. 2 Expanded Cpmmittee on Establishment of Guidelines for Proper Conduct of Animal
Experiments
Chairman: Ichiro Kanazawa (President, National Center of Neurology and Psychiatry)Vice-chairman: Hideaki Karaki (Professor Emeritus of The University of Tokyo)
Secretary: Setsuo Hirohashi (Director, National Cancer Center Research Institute)
Secretary: Izumi Washitani (Professor of the Graduate School of Agricultural and Life Sciences, University ofTokyo) 㻌
Makoto Asashima (Second Division, Vice President of Science Council of Japan, Professor, University of Tokyo)
Noriko Osumi 䠄Professor of Developmental Neuroscience, Center for Translational and Advanced Animal Research(CTAAR), Tohoku University Graduate School of Medicine䠅
Masaru Taniguchi 䠄Director, RIKEN Research Center for Allergy and Immunology䠅
Akio Nomoto (Professor, Department of Microbiology, Graduate School of Medicine, University of Tokyo
Yasushi Miyashita (Department of Physiology, Professor and Chairman, University of Tokyo School of Medicine)
Hideo Yano 䠄Professor, Graduate School of Agriculture, Kyoto University䠅
Yasuo Ohno (Deputy Director General, National Institute of Health Sciences)
Hiroshi Ozaki (Professor, Professor of the Graduate School of Agricultural and Life Sciences, University of Tokyo)
Naoko Kagiyama (Senior Scientist, Central Institute for Experimental Animals)
Yoshikazu Shinoda (Professor, Tokyo Medical and Dental University, School of Medicine)
Norikazu Tamaoki (Deputy Director, Central Institute for Experimental Animals)
Ken-ichi Yagami (Director, Laboratory Animal Resource Center, University of Tsukuba)
Observers
Hiroki Matsuo (Director, Life Sciences Division, Research and Promotion Bureau, Ministry of Education, Culture,
Sports, Science and Technology
Noriyo Yoshikawa 䠄Deputy Director㻌 Health Sciences Division㻌 Minister's Secretariat, Ministry of Health, Laborand Welfare䠅
Katsuhiko Shoji 䠄Director, Office of Animal Companionship Policy and Coordination Division, Nature
Conservation Bureau, Ministry of the Environment

18