Future University – Hakodate Operation Procedure in Animal Experiments
September 15，2006
Decision at the Faculty Meeting(Purpose)
Article 1
This procedure provides the necessary information on animal experiments operated based on the“Future University – Hakodate Guidelines for Animal Experiments” (September 15, 2006;
Decision at the Faculty Meeting; hereinafter referred to as the “Guidelines”).
(Facilities)
Article 2
The facilities mentioned in Article 4 of the Guidelines are as follows.
(1) The facilities for the operation of animal experiments are laboratories 255 and 355, and therange of animal experiments must be as follows.
(2) Laboratories must have proper floors so that the stains from excretory substances and blood can
be cleansed and disinfected. In addition, local exhaust ventilation must be equipped foradequate ventilation.
(3) The control staff is responsible for maintaining good hygiene and preventing escape of animals.
The facilities must be organized so that experimental animals can be caught when they try toescape.
(Management and Carrying in of Experimental Animals)
Article 3
Points of concern regarding management and carrying in of experimental animals mentioned inArticle 6 of the Guidelines are as follows.
(1) Procurement of experimental animals
1) Experimental animals must be obtained legally.
2) The transfer of recombinant DNA animals and specific adventives and the import ofexperimental animals belong to Rodentia or Monkeys must abide by the related laws.
3) The experimenter must use only the experimental animals produced to fit for the purpose, orto which the result of microorganism monitoring or infection examination is attached.
(2) Storage of the records of experimental animals
The experimenter must store the records of experimental animals properly, by clarifying the sourceof procurement of experimental animals as well as organizing the record files on the storage
environment.

(3) Management of cages
Considering the change in temperature and humidity inside of a cage, the following conditions mustbe fulfilled.
1) Cages must have the structure and the strength in accordance with the kinds of animals toprevent them from escaping.
2) Experimental animals must be able to maintain normal body temperature in a cage.
3) Experimental animals must be able to maintain proper posture for urination or defecation ina cage.
4) According to the behavior unique to each animal, the experimental animal must be kept
clean and dry all the time in a cage.
5) Cages must be safe for experimental animals. (Cages must have no sharp edge or projecting
part. Body and limbs of experimental animals are not stuck in the gap of the cage.)6) Cages must have the structure and the materials that allow to be cleansed, disinfected, andsterilized easily.
(Operation of an Experiment)
Article 4
Points of concern regarding operation of an experiment mentioned in Article 7 of the Guidelines are
as follows.
(1) Positioning of a body
Points of concern regarding positioning of a body (i.e. to partially or totally limit animals’ normal
body movement by using positioner to process various experiments) are as follows.
1) The positioner must be appropriate size and easy to operate. When using the positioner,
discomfort and injury to experimental animals must be minimized.
2) While positioning, the experimenter must constantly observe the experimental animal. When
external injury or deconditioning is found, the experimental animal must be released fromthe positioner immediately.
(2) Restriction of feeding
When feeding of experimental animals has to be restricted, the experimenter must clarify thescientific basis and make sure the animal is fed the minimum amount of food and water during anexperiment.
(3) Surgical procedure
If an animal experiment involves surgical procedure, the following points must be concerned tominimize pain of experimental animals within the range of the research purpose.
1) In addition to improve surgical skills, aseptic treatment during the experiment andpostoperative management must be done appropriately.
2) When non-aseptic body part such as gastrointestinal (GI) tract has to be surgically exposedor the immune function may be decreased due to such surgery, antibiotics must be given to
the experimental animal.
(4) Analgesia and anesthesia
1) When experimental animals show peculiar behavior unique to the species: such as crying,glooming, displaying abnormal facial expression or posture, or stopping moving;
appropriate analgesia must be given to the animals.
2) If necessary, the experimenter must consult doctors, veterinarians, and pharmacists forchoosing analgesia or anesthesia that does not undermine the purpose of research.
3) Suggested methods and dosages for anesthesia are as follows.
a) Injection method
 Pento-barbital sodium: dosage (mg/kg) (route, duration of action)
Mice: 30-50 (i.p., 30-60 min.)Rats: 30-40 (i.p., 30-60 min.)Guinea pigs: 20-30 (i.p., 30-60 min.)Rabbits: 25-30 (i.v., 30-60 min.)Dogs: 25 (i.v., 60 min.)Cats: 25-35 (i.v., 60 min.) Ketamine hydrochloride: dosage (mg/kg) (route, duration of action)
Rabbits: 15 (i.m., 20-30 min.)Dogs: 10-25 (i.m., 20-60 min.)Cats: 15-25 (i.m., 20-60 min.)Monkeys: 10-40 (i.m., 20-90 min.) Urethane: dosage for euthanasia (mg/kg) (route)Rabbits: 1.5 (i.p.)(Note)

i.p.: intraperitoneal injection, i.v.: intravenous injection, i.m.: intramuscular injectionb) Inhalation anesthesia
Ether: Mice, Rats, and Guinea pigs

Halothane: Dogs
(5) Humane Endpoint
Points of concern regarding humane endpoint are as follows.
(Humane endpoint: The timing to terminate an experiment by euthanasia of experimental animals torelease them from agony)
1) If necessary, humane endpoint must be set at the stage of planning of animal experiments.
2) When setting humane endpoint, the experimenter must refer to the related internationalguidelines.
3) The signs to apply humane endpoint are: difficulty of feeding; signs of agony (self-injury,abnormal posture, difficulty of breathing, or crying); long-term abnormal appearance
without a sign of recovery (scours, bleeding, dirty external genitals, etc.); rapid weight loss
(20% or less of the original body weight in several days); or greatly-increased tumor size
(10% or more of the original body weight).
(6) Euthanasia
1)

When euthanizing experimental animals (or inducing immediate unconsciousness and death
without causing pain), the experimenter must follow international guidelines as well as the“Guidelines Concerning the Disposal of Experimental Animals” (The 40th Notification ofPrime Minister’s Office on July 4, 1995).

2) Drugs and methods for euthanasia must be chosen properly in accordance with the species
and the purpose of the experiment.
3) Common methods for euthanasia are as follows.
Chemical methods: Excessive amount of barbiturate anesthesia, non-explosive
inhalation anesthesia, carbon dioxide gas
Physical methods: Bone dislocation of cervical vertebra, beheading, exsanguinations
under anesthesia(7) Consideration of safety management
When operating recombinant DNA experiments; animal experiments using radioactive materials orradioactive ray, experiments using poisonous/deleterious substances or psychopharmaceuticals, andanimal experiments using pathogens or toxic chemical compounds; the experimenter must refer tothe related laws and regulations.

[List of References]
1. Related laws, guidelines, etc.
(1) Law Governing Animal Protection and Control
http://www.env.go.jp/nature/dobutsu/aigo/amend_law2/index.html(2) Standards for Feeding, Management, and Pain Alleviation of Experimental Animals
http://www.env.go.jp/nature/dobutsu/aigo/anim guide/index.html(3) Guidelines Concerning the Disposal of Experimental Animals
http://www.env.go.jp/nature/dobutsu/aigo/anim_guide/index.html(4) The Ministry of Education, Culture, Sport, Science and Technology
Basic Guidelines Concerning the Operation of Animal Experiments at Research Institutions
http://www.mext.go.jp/b_menu/hakusho/nc/06060904.html(5) The Ministry of Health, Labor, and Welfare
Basic Guidelines Concerning the Operation of Animal Experiments at the Ministry of Health, Labor,and Welfare
http://www.mhlw.go.jp/general/seido/kousei/I-kenkyu/index.html(6) Law Concerning the Security of Diversity of Living Things due to the Restriction of the Use ofRecombinant DNA Organisms
http://www.maff.go.jp/carta/15hou97.pdf(7) Law Concerning the Prevention of Damages to Ecology by Specific Adventives
http://www.env.go.jp/nature/intro(8) Law Concerning the Prevention of Infection and the Medical Practice for Patients with Infection
http://www.acc.go.jp/mlhw/mhw_kansen_law/114.htm(9) Livestock Infection Prevention Law
http://law.e-gov.go.jp/htmldata/S26/S26HO166.html(10) Rabies Prevention Law
http://law.e-gov.go.jp/htmldata/S25/S25HO247.html(11) The Head of the Science and International Affairs Bureau, Ministry of Education; Animal
Experiment at Universities (Announcement); 1987(12) Research Promotion Bureau, Ministry of Education, Culture, Sport, Science and Technology;
Introduction of Animal Experiment at Universities (Announcement); 2001(13) Science Council of Japan; Formulation of Guidelines for Animal Experiment(Recommendation); 1980
(14) Science Council of Japan; Report by the Special Committee on the Development of Life
Science and Formation of Social Consensus; Handling of Animals for Education and Research; 1997(15) Report at the Science Council of Japan; Improvement of Social Understanding on Animal
Experiments (Proposal); 2004

http://www.scj.go.jp/ja/info/kohyo/pdf/kohyo-19-t1015.pdf(16) Science Council of Japan; Guidelines for the Proper Operation of Animal Experiments; 2006
http://www.scj.go.jp/ja/info/kohyo/pdf/kohyo-20-k16-2.pdf(17) Science Council, Ministry of Education; Specific Research Area Promotion Section; Bioscience
Division; Safety Management Manual for Laboratory Microorganisms at Universities (Draft); 1998
2. Description of related laws, textbooks, etc.
(1) Research Group of the Law Governing Animal Protection and Control; Companion to theBusiness on Animal Protection and Control; Taisei-shuppan (publisher); 2006
(2) Research Group of the Animal Protection Theory; Q & A Concerning the Revised Law
Governing Animal Protection and Control; Taisei-shuppan (publisher); 2006
(3) Research Group of Breeding and Management of Experimental Animals; Description of theCriteria Concerning Breeding and Management of Experimental Animals; Gyousei (publisher); 1980
(4) Expert Committee Dealing with the Disposal of Experimental Animals; Description of theGuidelines Concerning the Disposal of Experimental Animals; Japan Veterinary MedicalAssociation; 1996
(5) Animal Experiment Facilities Council of National University; Safety Measures for InfectedAnimal Experiments; 2001
(6) Animal Experiment Facilities Council of National University/Animal Experiment Facilities
Council of Public and Private Universities; Guidelines for the Prevention of Hemorrhagic Fever withRenal Syndrome at Universities; 2001
(7) Architectural Institute of Japan; Architecture and Equipments of Facilities for ExperimentalAnimals; Adthree (publisher); 1996
(8) Japanese Society of Laboratory Animals; Skills and Application Involved in Experimental
Animals - Introduction and Practice; Adthree (publisher); 2004
(9) Japanese Society of Laboratory Animals; Monitoring Manual for Laboratory Microorganisms;
Adthree (publisher); 2005(10) Manual for Handling Experimental Animals with Infection, supervised by Kazuyoshi
Maeshima; Adthree (publisher); 2000
3. International/Overseas guidelines and textbooks(1) 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)(2) ICLAS International Council for Laboratory Animals). Guidelines for the regulation of animalexperimentation. 1974

(3) CIOMS involving animals, 1985. (Council for internationa1 0rganizations of MedicalSciences)/WHO. international guiding principles for biomedical research(4) ILAR (Institute for Laboratory Animal Research)-NRC (National Research Council). Guide forthe care and use of laboratory animals (7th edition). National Academy Press. 1996.
(Japanese Translation: 鍵山直子，野村達次監訳.実験動物の管理と使用に関する指針ソフトサ
イエンス社.1997.)(5) European Communities (Presently: European Union). Council Directive 86/609/EEC. 1986.
(Japanese Translation: 福井正信監訳.EC(欧州共同体)動物実験指針.ソフトサイエンス社.1991.)(6) SCAW (Scientists Center for Animal Welfare). Categories of biomedical experiments based onincreasing ethical concerns for non-human species.1987.(解説 国立大学動物実験施設協議会，動物実験処置の苦痛分類に関する解説.2004.)(7) CCAC (Canadian Council on Animal Care). Guidelines on choosing an appropriate endpoint inexperiments using animals for research teaching and testing 1997.
(8) ILAR一NRG Humane endpoints for animals used in biomedical research and testingiLAR Journa1 41(2) Special edition, 2000
(9) UK Co-ordinating Committee on Cancer Research UKCCCR guidelines for the welfare ofanimals in experimental neoplasia. 1988.
(10) AVMA (American Veterinary Medical Association). 2000 report of the AVMA panel oneuthanasia JAVMA 218 (5), 669-699(11) Working Party of the European Commission. Recommendation for euthanasia of experimental
animals Part l. Laboratory Animals 30, 293-316, 1996 & Part 2. Laboratory Animals 31,1-32, 1997(12) EFPIA (European Federation of Pharmaceutical industries Associations) & ECVAM (EuropeanCentre for the Validation of Alternative Methods). A good practice guide to the administration ofsubstances and removal of blood, including routes and volumes 1999
(13) Frecknell P Laboratory animal anesthesia (2nd ed). Academic Press. 1996. (JapaneseTranslation: 倉林議監修.ラボラトリアニマルの麻酔―げっ歯類・犬・猫・大動物一.学窓.1998.)(14) Smith & Swindle (eds.). Research animal anesthesia and surgery. SCAW. 1994
(15) IATA (international Air Transportation Association. IATA live animal regulations. 1995. (22ndedition).
(16) 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. (Japanese Translation: 倉田毅訳. CDC・NIH.納庄物学・医学実験室のバイオセーフティ.第3版。医朝1996.)(17) U.S. National Research Council. Occupational Health and Safety in the Care and of

ResearchAnimais.1997. (Japanese Translation: 日本実験動物環境研究会編(黒澤努`佐藤浩監
訳). 実験動物の管理と使用に関する労働安全衛生指針.アドスリー.2002.
(18)Hau J&van Hoosier, Jr. G Hand book of laboratory animal science (2nd ed), Vol.l Essential
principles and practices, Vol. 2 Animal models CRC Press. 2003,