Policy and Regulation for the Care and Use of Laboratory Animals
July 30, 2003
This guide states policies and regulation for planning and performing animal experiments or protocols at Seoul National University. Seoul National University is committed to conduct appropriate care and use of animals for a more ethical and scientific test to perform better management on authorization of animal experiment plans, acquisition of laboratory animals, nurturing and releasing them upon each experiment.
II. (The Scope of Application)
This guide applies to researches, experiments, tests or education on live vertebrate animals within Seoul National University campus.
III. (Protocol Committee)
① In order to review and decide upon articles of regulation regarding humane care and scientific use of laboratory animals, Protocol Committee (referred as to “Committee” hereinafter) is established and operated. The Committee is consisted of three or more members that must include a veterinarian with professional experience and related education on animal experiments and a non-affiliated member who is generally interested in use and management of laboratory animals.
② The Committee reviews all the researches, experiments, tests or education on live vertebrate animals within Seoul National University campus. It reserves a right to request a revision or modification on protocol s. It also has a right to defer an authorization on or terminate experiments.
③ A biweekly meeting is held. The number of total attendance at each meeting must be equal to or more than half of the Committee members which must include one non-affiliated member.
④ The Committee must survey and evaluate all the animal facilities on campus at least once in six months. Then, it must submit a report to the president of university to verify if laboratory animals are humanely used for researches and education.
IV. (The Beginning and Completion of Protocol s)
① One must make consideration on substantial care of animals to minimize termination of animals by setting up an optimal protocol to meet purpose of experiment and using means of alternatives.
② An experiment operator must choose laboratory animals based on their breeds, pedigrees, weight, sex, groups, hereditary and microbiological quality and surroundings.
③ One must file a protocol and receive an approval from the Committee for performing all the protocol s including of vertebrate animals upon its careful review.
④ When entering into an agreement, in which a protocol is part of, on either an on-campus or outside research fund, President (Dean of Research Affairs, Dean of each College at SNU) verifies the Committee’s authorization number regarding the planning report including the protocol and approves it.
⑤ In case of any modification or amendments in the experiment details, one must submit a statement of modification to the Committee chair. Upon the completion of experiment, a protocol completion report must be submitted to the chair.
V. (Management and Care of Laboratory Animals)
① The person in charge of management and care of laboratory animals and facilities, experimenters and breeders must control ventilation, illumination, noise, temperature and humidity of the facilities based on biological traits of each animal and choose food, drinking water and bedding. In addition, they must repair and maintain the facilities in order to help properly conduct protocol s.
② Laboratory animals must be managed and cared in a facility approved by the Committee.
VI. (Veterinary Management of Laboratory Animals)
① A Committee-approved researcher may request for a transfer of laboratory animals to an administrative staff of the Committee. The administrative staff will provide laboratory animals upon consultation to a veterinarian of the laboratory animal facility.
② Experimenters and breeders must healthy animals only when taking in laboratory animals while observing the condition of animals and keeping transfer records.
③ Experimenters and breeders must make daily observation on growth progress, development and condition of entire body of laboratory animals. If necessary, they may examine presence of contraction of a disease. In addition, they must receive consultation and directions from a veteran if special knowledge or skills of a veterinarian are required in a periodical microbiological monitoring or execution of an experiment.
④ Experimenters must consider precision in experimental methods to achieve results from an experiment while minimizing the number of terminated animals.
VII. (Termination of Laboratory Animals Upon Completion of an Experiment)
① Upon the completion of an experiment or for those animals that are necessary to drop out of an experiment, experimenters must induce their loss of consciousness and perform euthanasia to let them flee from pain as quickly as possible.
② If a laboratory animal dies while breeding, in an experiment or upon the completion of experiment, the experimenter must keep the dead body in a carcass storage in order not to contaminate the surroundings and affect physical well-being of human and other animals.
③ Each laboratory animal facility on campus must incinerate dead animals based on the standard on contaminating waste and make a report to the Committee every six months.
VIII. (Experiments Requiring Special Attention Including Safety Management)
① When conducting protocols dealing with physically, chemically and biologically hazardous materials (e.g., a radioactively isotopic element, pathogenic agent, recombinant DNA, carcinogenic agent, mutating substance and all other unidentified substances that is not met by the safety standard), one must receive an authorization from the Committee and conduct the experiment based on a separate rules and regulations for safety.
② Experimenters of protocol s must set necessary means of regulations in order not to contaminate human and animals and prevent any hazard by limiting the entry into an experimental area for those who are not participating in the experiment.
③ Experimenters must abide the Committee’s supervision when conducting protocol s concerning transgenic animals. In order to keep the security for the experiment, they must prevention methods on escape based on each animal’s habit in laboratory and facility.
Manual for the Use and Management of Laboratory Animal
Background for the Animal Experiment
An animal experiment is an experiment using laboratory animals of which purpose is to observe how animals react to certain experimental measures and further predicts how such a result will affect human or other animals. For example, new chemical and functional food substances including medicines, daily necessaries, cosmetics, disinfectants and agricultural chemicals, are tested through animal experiments to predict how they will have an effect on human before they are available to the general public. In addition, in order to develop a method of medical treatment or cure for an incurable disease, animals in similar condition of those humans with a certain disease are produced via physical, chemical and biological techniques. Such an animal experiment makes it possible to develop new medicines and forecast a medical treatment method for contamination accident and environmental pollution due to chemical substances.
A Researcher’s Attitude Towards the Animal Experiment
Russell and Burch proposed 3R’s for a researcher’s aims to maintain his/her integrity. In other words, they introduced the ideas of reduction and replacement of laboratory animals, and refinement of the animal experiment.
It is possible to reduce the number of laboratory animals if one carefully investigates any unnecessary experiments among all the testing performed up to date. A good example for such an attitude is that the number of animals currently used in an experiment finding 반수치사량, the very first testing among all clinical demonstrations of drugs and chemical substances, has significantly been reduced as opposed to the number in the past.
Some research groups, on the other hand, are actively searching for replacement for animals. Originally using animals have developed experimental zoology as replacement of human. If a better suited entity for an experiment than animals appears, it will be definitely possible to induce such an entity rather than the currently used laboratory animals. For that reason, the usefulness of invertebrates, cultivated cells and computer simulations were once examined as replacement experimental subjects. However, these entities could not satisfy both quantitatively and qualitatively due to their limited dissection conditions or physiological functions. Thus, the needs for animal experiments in these days are quite unavoidable that will last for a while to advance medicine as well as science.
In order to refine animal experiments, it is essential to improve the quality of laboratory animals and stabilize their reaction by upgrading housing surroundings. Then, it will secure reproduction of experimental results that will prevent us from duplicating experiments over and over again. In addition, increasing interoperability of testing data by standardizing laboratory animals and various testing methods can lead to the refinement of animal experiments. One of the most important aspects in the refinement of animal experiments is to minimize the pain on the laboratory animals. Termination upon the completion of experiment gives not only pain to animals, but also distress including anxiety, unpleasantness and despair especially to those with somewhat higher intelligence. Such a measure for pain on each laboratory animal is difficult since receptivity and expressions against pain are different for each type of the animal. However, if it were a condition for a person to feel pain, it would be the same for animals. Laws and regulations on the laboratory animals have been constituted based on the above apprehension at least. Thus, animal experiments can be classified based on the degree of pain on animals and many advanced nations have chosen such a method to authorize them. If an animal experiment is performed in the breeding period, anesthesia must be applied to minimize pain. Upon the completion of experiment, the method of euthanasia must be performed. Moreover, breeders must enable the laboratory animals to maintain good conditions through the veterinary knowledge and the use of such technique even in the daily management of these animals. In order to reach the two aims, which are the welfare of laboratory animals and scientific system for laboratory animals, the more expertized knowledge and rigid legal regulations are necessary.
1. The housing facilities for laboratory animals at Seoul National University are distributed to three separate locations, Kwanak, Yeongun and Suwon campuses. This policy applies to all the animal experiments at Seoul National University’s laboratory animal facilities stated in the appendix 1.
Appendix 1 Seoul National University’s Laboratory Animal Facilities
2. Definition of the Animal Experiment
An ‘animal experiment’ can be defined as a scientific experiment on animals as an object. It includes researches (observation and experiments) to prove inquiries for facts or hypotheses, tests to search for make-up and effects on new substances and methods regarding human life, education to deliver bioscience knowledge and techniques for the current and next generations and extraction of raw materials to collect research or diagnostic materials and materials for medicine.
3. Target Animals
This policy defines target animals as vertebrate animals—that is, laboratory animals, livestock and wildlife animals. It excludes all the experiments on dead animals or parts of dead animals and wild animals without altering their surroundings.
1) Laboratory Animals
Laboratory animals are vertebrate animals used in researches, education, experiments and tests including Primates except human, such as dogs and cats; small rodents, such as guinea pigs, hamsters and mice; rabbits, birds, aquatic animals, reptiles and amphibians.
When using livestock for research, education and experiments, they are classified into biomedical and agricultural purposes. Such a distinction is clear in most researches. For example, animal models for human diseases, organ transplants and main surgical treatments are aimed at biomedical purposes and experiments and researches for production of meat, milk, eggs and textiles at agricultural purposes. However, these two distinct classifications often are not easily distinguishable in several experiments on nutrition and diseases. Housing system of livestock used in biomedical studies may be different from that of agricultural studies, but may not differ from one another. The livestock used in biomedical and agricultural researches are raised in cages, barns, small sheltered or outdoor housing. Some agricultural researches require certain conditions to minimize environmental variation and some biomedical researches take place in outdoor housing facilities. Thus, protocol rather than scope of researches determines research locations (outdoor housing, laboratory) as a standard.
The Committee decides on standards and definitions of the use and management of animals based on the user’s purpose, protocol and considerations on welfare of animals. Regardless of the scope of research application, research centers must supervise all the laboratory animals and minimize their pain and distress. This policy applies to all the livestock used in biomedical studies including typical animals living in outdoor housing facilities.
3) Wild Animals
When observing or using vertebrate animals in wilderness for biomedical or behavioral researches, basic regulations regarding humane use and protection do apply as well. The examiner at the scene of such studies must assure the Committee that his/her collection of specimen or intrusion into the environment follows this regulation. The Committee must examine the scene to guarantee the safety and well-being for the researchers of the scene and other animals.
1. Organization of the Committee and Its Authority
The president of the university must organize the Protocol Committee to enable this policy to be logically put in practice, to give proper advice and supervision to researchers and to provide administrative support for the committee to appropriately work. The committee must be consisted of at least three members without a separate committee for each college that the central committee supervises.
The Committee is responsible for supervision and evaluation on the program for the management and use of animals. Its function includes inspection on facilities, evaluation on the program and animals’ sphere of activity, submission of reports from each facility supervisor, examination on the use of suggested animals in researches, experiments and education (protocol), establishment on the receipt system and interest of each facility’s in the management and use of animals.
The Committee must hold meetings as frequently as possible to perform its duties and a meeting must be held at least once in two weeks. Minutes of the Committee meetings and results from meetings must be kept for five years. It must examine the animal management program at least once in six months and inspect animal facilities and their sphere of activity. A report upon such examination and inspection with majority approval of the Committee must include conditions of the program for the management and use of animals and other activities that must be submitted to the president.
The Committee is comprised of non-scientific/lay/non-affiliated members as well as scientific/affiliated members, to ensure a balance of professional expertise and community perspective. All Committee members must participate in the review process and a biweekly meeting will be held on Mondays. A fixed number for each meeting must be equal to or greater than one half of regular voters where at least one non-affiliated member must be present. The Committee members must perform a follow-up inspection at least once a year besides the regular check-ups where they meet with researchers and staffs of animal facilities to verify accordance between the use of animals and protocol; drugs and materials used in protocol s and animal records.
2. The Committee’s Confidentiality
The Committee members must not use information from their committee activities for the personal benefit or drains of such information to any others. Those articles requiring confidentiality are related to an agreement of researchers, research process and progress, statistical information that requires the security of research facilities, researchers’ source of income, profit and loss and sales. The above information requires confidentiality. If anyone breaks this rule, (1) he/she will be removed from the Committee and (2) will be responsible for civil liability according to the pledge at the time of his/her appointment to the Committee.
3. Submission of the Protocol
For a protocol using vertebrate animals, a protocol form must be completed and filed before its beginning phase that must be approved by the Committee regardless of types of animals and their purpose.
Appendix 2. Animal Use Protocol Form
4. Review for the Protocol
The review and approval process on a protocol begins with the completion of the form. Consultation with each facility’s veterinarian in charge is a prerequisite prior to the submission of protocol. The review of Committee is progressed in three different forms, which are regular, expedited and executive reviews. The review option is determined upon predicted pain on and anxiety of animals and protocol methods.
Appendix 3. Classification of Protocols Upon the Pain on Animals
A review is carried out with the following steps.
Step 1: Submission of the Protocol. Investigators submit completed protocols to an administrative staff at the Committee office.
Step 2:Initial Review. Upon the submission of the protocol, a type of review will be determined upon predicted pain on and anxiety of animals and protocol methods. Modification on the protocol may be examined here.
Step 3:Review. At a biweekly meeting, the Committee considers new protocols requiring full committee review, reviews the expedited reviews and executive reviews. Possible outcomes of the Committee’s review include unqualified approval, approval pending modification(s) and/or clarification(s), table (deferral) or.
a.Full Review- Two members from the Committee are assigned. The primary reviewer is always a scientific representative of the Committee and is responsible for presenting a summary of the protocol at the meeting along with any concerns or points requiring clarification or modification. The secondary reviewer adds any additional concerns. The protocol is then open for discussion by the Committee.
b.Expedited Review- One reviewer is assigned. The reviewer is give three days to complete the review and the reviewer’s comments and the protocol are then forwarded to the remainder of the Committee for their review. If no further comments are received and there is no request for a full committee review, a letter requesting clarifications and/or modifications is sent to the investigator or if a recommendation to approve outright has been made, a Verification of Approval is sent.
c.Executive Review- If the purpose, hypothesis and laboratory animals used in the experiment are the same as the previously approved protocol—in other words, if it were a review on a continuous project, the Committee chair alone reviews and approves protocols falling in this category. This category includes continuing reviews, previously approved protocols that have been resubmitted or identical protocols submitted to different funding agencies, protocols with no direct animal use, e.g. funds will be used for the use of shared animal products or slaughterhouse materials or salary support for other researchers.
Step 4:Investigator Notification. If a protocol receives unqualified approval, the investigator is provided with a Verification of Approval and letter certifying the approval. In cases where the Committee requires clarification(s) or modification(s), the investigator is notified by the Committee chair in a letter. In such cases, the approval is issued following receipt of an acceptable response from the investigator. In cases of deferred or disapproved protocol, the investigator is notified by the chair and advised as to available options.
5. Criteria for Review
Protocols may contain potential procedures that may potentially cause inexperienced or uncontrollable pain or distress. Such procedures may include physical restraints, multiple surgeries, limits to the access of food and water, the use of adjuvants, termination upon the completion of protocols, use of hazardous stimuli, stimulation on skin and the cornea, induction of excessive sized tumor, blood-collecting from the heart or suborbital sinus and housing in an abnormal environment. General information on research methods and purposes must be searched from texts, other scholars or references that informs of influences of such studies on animals. If an experiment is performed via an unknown method, a qualified preliminary examination is advised to be carried out under the supervision of a Committee member to estimate possible influences and outcomes on animals.
Activities – All activities involving animals must be in accord with this regulation and policy for the welfare of animals.
Pain/Distress – Must avoid/minimize discomfort/distress/pain. If pain/distress is caused, appropriate sedation, analgesia or anesthesia will be used. Use of paralytics is prohibited. Animals with chronic/severe unrelievable pain will be painlessly killed.
Alternatives – The protocol investigator considers alternatives to procedures that may cause more than momentary or slight pain or distress to the animal and has provided a written narrative description of the methods and sources. These alternatives include methods that interferes less with animals, selecting another breed, cultivating cell or organs and the use of computer simulation.
Rationale and Methods – All proposals must include:
Identification of the species and the approximate number of animals to be used;
A rationale for involving animals and for the appropriateness of the species and numbers of animals to be used;
A description of procedures designed to assure that discomfort and pain to animals will be limited to that which is unavoidable for the conduct of scientifically valuable research, including provision for the use of analgesic, anesthetic, and tranquilizing drugs where indicated and appropriate to minimize discomfort and pain to animals; and
A description of any euthanasia method to be used.
Duplication – Must provide assurance that activities do not unnecessarily duplicate previous efforts.
Surgery – Must meet requirements for sterile surgery and pre/post operative care. Cannot use one animal for several major operative procedures from which it will recover, without meeting specified conditions.
Euthanasia – Euthanasia method must be consistent with this policy that is in a humane way.
Housing/Health – Animal living conditions must be consistent with standards of housing, feeding and care directed by veterinarian or scientist with appropriate expertise. Medical care must be provided by qualified veterinarian.
Qualifications – Personnel must be appropriately trained and qualified.
Safety of the Working Environment for the Personnel – In case of protocols dealing with hazardous materials, safe environment and safety standards for the personnel must be provided.
Continuing Review Process – Upon the recommendation of the reviewer, the protocol is approved outright for period of time up to three years, which the Committee reviews at least once a year.
Three-Year Renewal – At the end of the third year of a protocol, the investigator must resubmit it for the Committee in order to continue research activities. A new Animal Use Protocol Form must be submitted; this form undergoes the same review process as any new protocol.
Modifications or Amendments to Approved Protocols – Modifications to approved protocols must be documented appropriately, reviewed, and approved. The method for obtaining approval for a modification or amendment is similar to that for a complete protocol. A letter of requesting the modification including an explanation of the rationale for the change, any amended Animal Use Protocol pages resulting from this change should be submitted to the Committee. The chair, in consultation with the veterinarian of each facility if necessary, determines if the modification is “minor” or “significant.” Minor modifications may be approved administratively by the chair and the veterinarian of each facility without full review. A major modification may entail a large change in numbers of animals being used or requested, an increase in invasiveness, a change in species, an increase in pain or discomfort, or a change in the method of euthanasia. Major modifications require review by the full committee. A written description of the significant changes in the protocol is provided to the Committee.
Termination of Protocols – It is the responsibility of the investigator to notify the Committee when a project is completed. Projects that have been completed, withdrawn or terminated are closed immediately upon notification. The administrative office of the Committee is notified by the chair of the all closures. All closed projects are filled in the administrative office of the Committee for a five-year period from date of closure.
An investigator must file a protocol and receive an approval from
the Committee in order to perform all animal experiments using vertebrate
2. Animal Care and Use Protocols
An investigator should plan for a protocol before an animal experiment that should be considered carefully to perform it scientifically. It is necessary to make an effort to seek a consultation with an animal experiment expert in order to achieve an appropriate animal experiment. An investigator must take considerations on selection of appropriate animals for the protocol’s purpose, the number of animals used, genetic and microbiologic quality and breeding conditions. Especially, an investigator must comply with each facility’s policy on microbiologic quality.
3. Physical Restraint
Physical restraint is the use of manual or mechanical means to limit some or all of an animal’s normal movement for the purpose of examination, collection of samples, drug administration, therapy, or experimental manipulation. Animals are restrained for brief periods, usually minutes, in most research applications. Restraint devices should be suitable in size, design, and operation to minimize discomfort or injury to the animal. Many dogs, nonhuman primates, and other animals can be trained, through use of positive reinforcement, to present limbs or remain immobile for brief procedures.
The following are important guidelines for restraint:
- Restraint devices are not to be considered normal methods of housing.
- Restraint devices should not be used simply as a convenience in handling or managing animals.
- The period of restraint should be the minimum required to accomplish the research objectives.
- Animals to be placed in restraint devices should be given training to adapt to the equipment and personnel.
- Provision should be made for observation of the animal at appropriate intervals, as determined by the Committee.
- Veterinary care should be provided if lesions or illnesses associated with restraint are observed. The presence of lesions, illness, or severe behavioral change often necessitates temporary or permanent removal of the animal from restraint.
4. Multiple Major Surgical Procedures
Major surgery penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic function. Multiple major survival surgical procedures on a single animal are discouraged, but may be permitted if scientifically justified by the user and approved by the Committee. For example, multiple major survival surgical procedures can be justified if they are related components of a research project, if they will conserve scarce animal resources, or if they are needed for clinical reasons. If multiple major survival surgery is approved, the Committee should pay particular attention to animal well-being through continuing evaluation of outcomes. Cost savings alone is not an adequate reason for performing multiple major survival surgical procedures.
5. Food or Fluid Restriction
When experimental situations require food or fluid restriction, at least minimal quantities of food and fluid should be available to provide for development of young animals and to maintain long-term well-being of all animals. Restriction for research purposes should be scientifically justified, and a program should be established to monitor physiologic or behavioral indexes, including criteria (such as weight loss or state of hydration) for temporary or permanent removal of an animal from the experimental protocol.
6. Selection of Animal Species·Pedigree·Quality
1) Selection of Animal Species
Selection of laboratory animals varies depending on the purpose and details of animal experiments; animals should be selected with consultation to references or experts. Generally, the objective of most researches is on examining principles regarding life phenomena so that it is desirable to choose appropriate model animals for such examination. Though a new animal may be selected for various inspection and safety tests, many animals have been chosen in accordance with regulations. Thus, they can be selected based on the existing regulations. There are frequent occasions that a set of methods is established for extraction of diagnostic or medical materials. It is imperative to select animal species for educational purposes to reach its fullest effect .
2) Selection of Pedigree
In terms of genetics, biological reaction of laboratory animals including response to medication, resistance and immunity vary based on species, pedigree, and sexes even in the same animal specie. Therefore, it is recommended to warrant genetic quality. In addition, infecting pathogenic agent of laboratory animals in microbiological point of view, even if it were an unapparent infection, not only manifests symptoms by experimental measures, but also confuses outcomes of animal experiments and spreads infections to healthy animals. Investigators must receive laboratory animals that come with the microbiologically monitored origin of animal species or quarantine certificates and follow facility manager’s manual on infection prevention as well.
Appendix 4. Classification of Laboratory Animals In Accordance With Microbial Control
3) Genetic Characteristics of Laboratory Animals
Genetic characteristics are important in regard to the selection and management of animals for use in breeding colonies and in biomedical research. Inbred strains of various species, especially rodents, have been developed to address specific research needs. The homozygosity of these animals enhances the reproducibility and comparability of some experimental data. Several methods of monitoring have been developed that use immunologic, biochemical and molecular techniques. Appropriate management systems should be designed to minimize genetic contamination resulting from mutation and mismating.
Transgenic animals are considered as unique resources. Care should be taken to preserve such resources through standard genetic-management procedures, including maintenance of detailed pedigree records and genetic monitoring to verify the presence and zygosity of transgenes. Cryopreservation of fertilized embryos, ova, or spermatozoa should be considered to safeguard against alterations in transgenes over time or accidental loss of the colony. Accurate recording, with standardized nomenclature where it is available, of both the strain and substrain or of the genetic background of animals used in a research project is important. Several publications provide rules developed by international committee for standardized nomenclature of outbred rodents and rabbits, inbred rats, inbred mice and transgenic animals.
Appendix 5. Nomenclature of Laboratory Animals
1. Personnel Qualifications and Training
1) All the personnel, who are related to animal experiments, must attend the training sessions sponsored by the Committee in order to be familiar with the purpose of this policy to perform experiments in near future.
2) The Committee must distribute qualified personnel to animal management, assistance position to animal experiment and other positions with wide knowledge, techniques, and experience on animal management and assistance to animal experiment. An animal facility technician must possess either the first or second class laboratory animal technician license certified by the Korean Association for Laboratory Animal Science.
2. Animal Environment, Housing and Management
Proper housing and management of animal facilities are essential to animal well-being, to the quality of research data and teaching or testing programs in which animals are used, and to the health and safety of personnel. Especially the environment (temperature, humidity, air stream, illumination, noise, odors, vibration, bedding, cage, water, food, housing, other animals in the same cage, personnel and so forth) heavily influences on the result of research data so that it is important to control these conditions. Thus, many factors should be considered in planning for adequate and appropriate physical and social environment, housing, space, and management. These include the following.
1) Housing Facilities
(1) Housing facilities must be a reported area upon receiving proper management and any temporary animal experiments are not allowed in these areas.
(2) The primary enclosure must consider temperature, humidity, ventilation, air stream, odors and illumination for well-being of animals. Following is the recommended standard for the environment based on the laboratory animal facility standard.
(3) It is recommended to separate animals based on the species and their microbiological characteristics. The reason for this is to reduce anxiety due to possible conflict among different species and infection from specie to others with different microbiological traits.
(4) It is essential for personnel and investigators’ breeding technique to establish proper working environment and administration in order to prevent a person from spreading infections. Especially cautions are required in housing management and disposal of animals with different biological characteristics.
2) Animal Environment
(1) Social Environment
Consideration should be given to an animal’s social needs. The socially environment usually involves physical contact and communication among members of the same species (conspecifics) although it can include noncontact communication among individuals through visual, auditory, and olfactory signals. When it is appropriate and compatible with the protocol, social animals should be housed in physical contact with conspecifics. For example, grouping of social primates or candids is often beneficial to them if groups comprise compatible individuals. Appropriate social interactions among conspecifics are essential for normal development in many species. A social companion might buffer the effects of a stressful situation, reduce behavioral abnormality, increase opportunities for exercise, and expand species-typical behavior and cognitive stimulation. Such factors as population density, ability to disperse, initial familiarity among animals, and social rank should be evaluated when animals are being grouped. An understanding of species-typical natural social behavior will facilitate successful social housing.
However, not all members of a social species can or should be maintained socially; experimental, health, and behavioral reasons might preclude a successful outcome of this kind of housing. Social housing can increase the likelihood of animal wounds due to fighting, increase susceptibility to such metabolic disorders as atherosclerosis, and alter behavior and physiologic functions. In addition, differences between sexes in compatibility have been observed in various species. These risks of social housing are greatly reduced if the animals are socially compatible and the social unit is stable.
It is desirable that social animals be housed in groups; however, when they must be housed alone, other forms of enrichment should be provided to compensate for the absence of other animals, such as safe and positive interaction with the care staff and enrichment of the structural environment.
Animal activity typically implies motor activity but also includes cognitive activity and social interaction. Animals maintained in a laboratory environment might have a more-restricted activity profile than those in a free-ranging state. An animal’s motor activity, including use of the vertical dimension, should be considered in evaluation of suitable housing or assessment of the appropriateness of the quantity or quality of an activity displayed by an animal. Forced activity for reasons other than attempts to meet therapeutic or approved protocol objectives should be avoided. In most species, physical activity that is repetitive, is non-goal-oriented, and excludes other behavior is considered undesirable.
Animals should have opportunities to exhibit species-typical activity patterns. Dogs, cats, and many other domesticated animals benefit from positive human interaction. Dogs can be given opportunities for activity by being walked on a leash, having access to a run, or being moved into another area (such as a room, larger cage, or outdoor pan) for social contact, play, or exploration. Cages are often used for short-term housing of dogs for veterinary care and some research purposes, but pens, runs, and other out-of-cage areas provide more space for movement, and their use is encouraged. Loafing areas, exercise lots, and pastures are suitable for large farm animals, such as sheep, horses and cattle. Considering a proper rack or cage is effective in simplifying care management operation or preventing from infection among animals. Cage heights should take into account typical postures of an animal so that its head will not touch the ceiling. At a minimum, an animal must have enough space to turn around and to express normal postural adjustments. It is recommended to provide an animal with bedding using materials of high elasticity. Recommended space for laboratory animals should follow the National Research Council (NRC) standards.
Appendix 6. Standards of Recommended Space for Laboratory Animals
3) Food, Water and Bedding
Animals should be fed palatable, noncontaminated, and nutritionally adequate food daily or according to their particular requirements unless the protocol in which they are being used requires otherwise. Animal facility managers should be judicious in purchasing, transporting, storing, and handling food to minimize the introduction of diseases, parasites, potential disease vectors (e.g., insects and other vermin), and chemical contaminants into animal facilities. They should urge feed vendors to provide data from feed analysis for critical nutrients periodically. The date of manufacture and other factors that affect shelf-life of food should be known by the user. Stale food or food transported and stored inappropriately can become deficient in nutrients. Careful attention should be paid to quantities received in each shipment, and stock should be rotated so that the oldest food is used first.
Areas in which diets and diet ingredients are processed or stored should be kept clean and enclosed to prevent entry of pests. Unused, opened bags of food should be stored in vermin-proof containers to minimize contamination and to avoid potential spread of disease agents. Exposure to temperatures above 21℃, extremes in relative humidity, unsanitary conditions, light, oxygen, and insects and other vermin hasten the deterioration of food. Precautions should be taken if perishable items—such as meats, fruits, and vegetables—are fed because storage conditions are potential sources of contamination and can lead to variation in food quality. Contaminants in food can have dramatic effects on biochemical and physiologic processes, even if the contaminants are present in concentrations too low to cause clinical signs of toxicity. For example, some contaminants induce the synthesis of hepatic enzymes that can alter an animal’s response to drugs.
Some experimental protocols might require the use of pretested animal diets in which both biologic and nonbiologic contaminants are identified and their concentrations documented. Most natural-ingredient, dry laboratory-animal diets that contain preservatives and are stored properly can be used up to about 6 months after manufacture. Vitamin C in manufactured feeds, however, generally has a shelf-life of only 3 months. The use of stabilized forms of vitamin C can extend the shelf-life of food. If a diet containing outdated vitamin C is to be fed to animals that require dietary vitamin C, it is necessary to provide an appropriate vitamin C supplement.
Refrigeration preserves nutritional quality and lengthens shelf-life, but food-storage time should be reduced to the lowest practical period and the recommendations of manufacturers should be considered. Purified and chemically defined diets are often less stable than natural-ingredient diets, and their shelf-life is usually less than 6 months; these diets should be stored at 4℃ or lower.
Autoclavable diets require adjustments in nutrient concentrations, kinds of ingredients, and methods of preparation to withstand degradation during sterilization. The date of sterilization should be recorded and the diet used quickly. Irradiated diets might be considered as an alternative to autoclaved diets.
Ordinarily, animals should have access to potable, uncontaminated drinking water according to their particular requirements. Water quality and the definition of potable water can vary with locality. Periodic monitoring for pH, hardness, and microbial or chemical contamination might be necessary to ensure that water quality is acceptable, particularly for use in studies in which normal components of water in a given locality can influence the results obtained.
Water can be treated or purified to minimize or eliminate contamination when protocols require highly purified water. The selection of water treatments should be carefully considered because many forms of water treatment have the potential to cause physiologic alterations, changes in microflora, or effects on experimental results. For example, chlorination of the water supply can be useful for some species but toxic to others.
Watering devices, such as drinking tubes and automatic waterers, should be checked daily to ensure their proper maintenance, cleanliness, and operation. It is better to replace water bottles than to refill them because of the potential for microbiologic cross-contamination; however, if bottles are refilled, care should be taken to replace each bottle on the cage from which it was removed.
Animal bedding is a controllable environment factor that can influence experimental data and animal well-being. The veterinarian or facility manager, in consultation with investigators, should select the most appropriate bedding material. No bedding is ideal for any given species under all management and experimental conditions, and none is ideal for all species. During autoclaving, bedding can absorb moisture and as a result lose absorbency and support the growth of microorganisms. Therefore, appropriate drying times and storage conditions should be used.
Bedding should be used in amounts sufficient to keep animals dry between cage changes, and, in the case of small laboratory animals, care should be taken to keep the bedding from coming into contact with the water tube because such contact could cause leakage of water into the cage.
(1) Cage – Periodic use of detergents or disinfectants are appropriate to maintain sufficiently clean surfaces. Not only cleaning cages and cage racks, frequent cleaning of feeders and watering device are recommended.
(2) Bedding Change – Bedding used in cages should be changed periodically to keep the cages clean since it keeps animals’ body surfaces dry. The frequency of bedding change for rats, mice, hamsters and other small rodents should be 1-3 times a week where dogs, cats and apes require daily bedding change.
(3) Cleaning and Disinfection of Cage Utensils – Periodic cleaning and efficient disinfection must be put in practice. Though sterilization through autoclaving is effective, plastic utensils should be washed with hot water then disinfected due to denaturalization from autoclaving.
5) Off Limits to Unauthorized Personnel – When transporting pathogenic agent to animal facilities or receiving contaminants from others besides facility personnel, trespassing of unauthorized personnel should be put in effect to prevent contamination and hazardousness. In addition, it is desirable to keep a record of those who come in and out of the animal facilities.
1. Animal Procurement
An investigator who has received an approval from the Committee, upon consultation with the attending veterinarian or animal facility manager, he/she should request assistance from an administrative staff of the Committee (Kwanak campus, College or Veterinary Medicine, College of Medicine) for animal procurement. The administrative staff of the Committee should verify the Committee’s authorization number, then make an order for all laboratory animals used on-campus from appropriate animal suppliers.
2. Veterinary Care
Veterinary medical care is essential part of an animal care to provide animals with preventive medical care and treatment, humane and ethical management of them, and to perform effective animal experiments accompanied with disposal. Especially it is inevitable to take consideration on animals not to cause unnecessary pain to them in terms of animal care and well-being. It is highly desirable to have an appropriate veterinary management under the supervision of an attending veterinarian with wide experience in laboratory zoology. In addition, one must keep in mind that medical examination and treatment on dogs, cats and other animals comply with the Veterinary License Act. Though such management is usually under the supervision of an investigator, sometimes it is necessary to receive consultation from facility managers or technicians.
1) Animal Procurement/Transportation and
(1) All animals must be acquired lawfully.
(2) An investigator and facility manager must confirm the means of animal transportation. It is important for them to observe and record the condition of animals’ health upon their arrival.
(3) Quarantine must be enforced to avoid providing unhealthy animals to experiments. Those newly received animals that will be used in experiments must be separated from the existing group of animals until they are proven to be healthy and acceptable for the experiments. The veterinary medical staff should evaluate the health, the pathogen status of newly received animals, and if necessary, treat any disease. For those animals from highly reliable suppliers, the above procedures can be substituted with their own pathogen monitoring test records. A careful quarantine is required for those dogs and cats caught from wilderness since their health status is not almost certain. This quarantine period can be a refinement period for these animals to adjust to a new environment; such a period helps them show stable physiological conditions and behaviors.
(4) Vaccination During the Quarantine Period – Preventive veterinary medical treatment including extermination of entozoa and epizoa must be carried out. Dogs must be registered with each region’s animal registration section according to the Rabies Prevention Act which must receive an antirabies serum injection once a year .
2) Surveillance, Diagnosis, Treatment, and Control of Disease
All animals should be observed for signs of illness, injury, or abnormal behavior by investigators, facility managers and technicians in cooperation with each other. In addition, it is imperative to observe conditions of food, water and cage utensils. As a rule, this should occur daily, but more frequent observations might be warranted, such as during postoperative recovery or when animals are ill or have a physical deficit. Unexpected death and sings of illness, distress, or other deviations from normal in animals should be reported promptly to the attending veterinarian or investigator to ensure appropriate and timely delivery of veterinary medical care. Animals that show sings of a contagious disease or injury should be treated immediately. Methods of disease prevention, diagnosis, and therapy should be those currently accepted in veterinary practice. Diagnostic laboratory services facilitate veterinary medical care and can include gross and microscopic pathology, clinical pathology, hematology, microbiology, clinical chemistry and serology. Microbial monitoring methods and the subject and microbe are indicated in Appendix 7. The choice of medication or therapy should be made by the veterinarian in consultation with the investigator. Examples of infectious agents that can be subclinical but induce profound immunologic changes or alter physiologic, pharmacologic, or toxicologic responses are Sendai virus, Kilham rat virus, mouse hepatitis virus, lymphocytic choriomenigitis virus, and Mycoplasma pulmonis. The principal method for detecting viral infections is serologic testing. Other methods of detecting microbial infections, such as bacterial culturing and histopathology and DNA analysis using the polymerase chain reaction, should be used in combinations that are most suitable for specific requirements of clinical and research programs. Transportable tumors, hybridoma, cell lines, and other biologic materials can be sources of munine viruses that can contaminate rodents. The mouse-antibody-production (MAP), rat-antibody-production (RAP), and hamster-antibody-production (HAP) tests are effective in monitoring for viral contamination of biologic materials and should be considered. Animals that show sings of contagious disease should be isolated from healthy animals in colony. If an entire room of animals is known or believed to be exposed to an infectious agent, the group should be kept intact during the process of diagnosis, treatment, and control.
Appendix 7. Microbial Monitoring
3. Consideration on Experiment Management
1) Degree of Pain on Animals
It is the investigator’s responsibility to recognize and minimize the degree of pain on animals in experiment management. However, it is not easy to be aware of such degree of pain. For example, the only way to learn scientific knowledge on sense of pain on animals is through studies of comparative anatomy, physiology or pathology as a basis. Thus, investigators must sympathize animals’ pain not only with knowledge from scientific studies, but also with humane perspectives. Furthermore, it is essential to take special considerations to reduce pain.
Appendix 3 shows classification of medical and biological experiments based on the degree of pain given to animals. Surely this is not a definite standard, but it is imperative to follow such index as a form of standards to place each experiment into a certain category. Investigators must do their best to perform their experiments as a lower categorized one.
2) Pain, Analgesia, and Anesthesia
The proper use of anesthetics and analgesics in research animals is an ethical and scientific imperative. Such use requires an effort to avoid or minimize anxiety, pain or distress. Unless scientifically proven, it is imperative to use appropriate amounts of sedatives, analgesics and anesthesia in tests that cause little ache or pain. Euthanasia might be necessary at the end of a protocol or as a means to relieve pain or distress that cannot be alleviated by analgesics, sedatives, or other treatments. The Committee gives an advice on the selection and use of necessary medications. Attending these animal tests without using such medication must follow the Committee’s judgment.
Fundamental to the relief of pain in animals is the ability to recognize its clinical signs in specific species. Species vary in their response to pain, so criteria for assessing pain in various species differ. Some species-specific behavioral manifestations of pain or distress are used as indicators, for example, vocalization, depression, or other behavioral changes, abnormal appearance or posture, and immobility. It is, therefore, essential that personnel caring for and using animals be very familiar with species-specific (and individual) behavioral, physiologic, and biochemical indicators of well-being. In general, unless the contrary is known or established it should be assumed that procedures that cause pain in humans also cause pain in animals.
The selection of the most appropriate analgesic or anesthetic should reflect professional judgment as to which best meets clinical and humane requirements without compromising the scientific aspects of the research protocol.
Some classes of drugs—such as sedatives, anxiolygtics, and neuromuscular blocking agents—are not analgesic or anesthetic and thus do not relieve pain; however, they might be used in combination with appropriate analgesics and anesthetics. Neuromuscular blocking agents (e.g., pancuronium) are sometimes used to paralyze skeletal muscles during surgery in which general anesthetics have been administered. When these agents are used during surgery or in any other painful procedure, many signs of anesthetic depth are eliminated because of the paralysis. However, autonomic nervous system changes (e.g., sudden changes in heart rate and blood pressure) can be indicators of pain related to an inadequate depth of anesthesia. If paralyzing agents are to be used, it is recommended that the appropriate amount of anesthetic be first defined.
Appendix 8. Recommended Anesthesia and Anesthetic Methods
Restraint is an act of restraining animals without causing them to fear or anxiety that helps investigators perform such experiments while animals remain motionless; it is also known as stationary.
Such a treatment minimizes pain given to animals significantly, makes experiment management easier and prevents investigators from any potential. 보정’s condition determines the outcome of experiment so that it is desirable to receive help from experienced technicians. However, a long-term restraint must be avoided; it is imperative to set the duration of such treatment. In case of using any restraint equipment, the investigator must be an expert of such equipment prior to the experiment.
4) Management of Surgery and Post Operative Care
A surgery must be performed by a well-experienced investigator or under his/her supervision. In survival surgery sufficient sterilization must be applied in the course of surgery and a properly trained person must monitor the animal with appropriate care upon the surgery. An appropriate amount of anesthesia should be a factor for a safe surgery so that precautions should be taken for the anesthetic management during the surgery and recovery upon the surgery.
Presurgical planning, personnel training, sterilized surgical techniques, animal care and appropriate precautions taken throughout the courses of the protocol will improve the outcome of the surgery. The influence of each factor may vary depending on the complexity of surgical procedures and/or animal species used in the surgery. A team approach towards surgical planning often brings successful outcomes with consultation from experts in different fields.
Euthanasia is the act of killing animals by methods that induce rapid unconsciousness and death without pain or distress. Unless a deviation is justified for scientific or medical reasons, methods should be consistent with the 2002 Report of the AVMA Panel on Euthanasia. In evaluating the appropriateness of methods, some of the criteria that should be considered are ability to induce loss of consciousness and death with no or only momentary pain, distress, or anxiety; reliability; nonreversibility; time required to induce unconsciousness; species and age limitations; compatibility with research objectives; and safety of and emotional effect on personnel.
Euthanasia might be necessary at the end of a protocol or as a means to relieve pain or distress that cannot be alleviated by analgesics, sedatives, or other treatments. Protocols should include criteria for initiating euthanasia, such as degree of a physical or behavioral deficit or tumor size, that will enable a prompt decision to be made by the veterinarian and the investigator to ensure that the end point is humane and the objective of the protocol is achieved. Euthanasia should be carried out in a manner that avoids animal distress. In some cases, vocalization and release of pheromones occur during induction of unconsciousness. For that reason, other animals should not be present when euthanasia is performed.
The selection of specific agents and methods for euthanasia will depend on the species involved and the objectives of the protocol. Generally, inhalant or noninhalant chemical agents (such as barbiturates, nonexplosive inhalant anesthetics, and CO2) are preferable to physical methods (such as cervical dislocation, decapitation, and use of a penetrating captive bolt). All methods of euthanasia should be reviewed and approved by the Committee. Death should be confirmed by personnel who can recognize cessation of vital signs in the species being euthanatized. Euthanatizing animals is psychologically difficult for some animal-care, veterinary, and research personnel, particularly if they are involved in performing euthanasia repetitively or if they have become emotionally attached to the animals being euthanatized. When delegating euthanasia responsibilities, supervisors should be aware of this as a potential problem for some employees or students.
Appendix 9. Methods for Euthanasia of Laboratory Animals
2. Waste Disposal
Infectious animal carcasses are transported and incinerated in accordance with the infectious waste disposal regulations at Seoul National University. Other wastes are disposed based on the methods of waste collection; if necessary there is disinfection of wastes. Storages to these wastes temporarily before the final collection must be kept separately from other storage areas; intrusion by flies, wild rats or other animals are not allowed.
Conventional, biologic, and hazardous waste should be removed and disposed of regularly and safely. There are several options for effective waste disposal. Waste containers should be leakproof and equipped with tight-fitting lids. It is good practice to use disposable liners and to wash containers and implements regularly. There should be a dedicated waste-storage area that can be kept free of insects and other vermin. If cold storage is used to hold material before disposal, a properly labeled, dedicated refrigerator, freezer, or cold room should be used.
Hazardous wastes must be rendered safe by sterilization, containment, or other appropriate means before being removed from the facility. Radioactive wastes should be maintained in properly labeled containers. Their disposal should be closely coordinated with radiation-safety specialists in accord with Seoul National University’s environmental safety investigator’s guide. Procedures for on-site packaging, labeling, transportation, and storage of infectious wastes including animal carcasses should be integrated into occupational health and safety policies. Hazardous wastes that are toxic, carcinogenic, flammable, corrosive, reactive, or otherwise unstable should be placed in properly labeled containers and disposed of as recommended by occupational health and safety specialists. In some circumstances, these wastes can be consolidated or blended.
1. Animal Experimentation Involving Hazards
Institutions should have written policies governing experimentation with hazardous biologic, chemical, and physical agents. An oversight process should be developed to involve persons, such as investigators, facility managers and technicians, who are knowledgeable in the evaluation of hazards and safety issues. It is especially important to have institutional rules for safety and master experimental methods. The Committee must supervise such particular experiments at Seoul National University carefully. Hazardous materials include radioactive materials, pathogenic agent, recombinant DNA, carcinogenic agent, mutating substance and all other unidentified substances that are not met by the safety standard. In addition, it is imperative to pay attention to bacteria or virus, and wild animals’ germs though these are not directly treated at the animal facilities.
Radioactive materials – The use of radiation must comply with the Korean law, Seoul
When injecting hazardous materials into an animal, investigators must examine its influence on the animal before the injection in terms of animal care. The hazardous materials must be sealed up in research facilities. In order to do so, research areas must be established, air roof and air filter must be equipped in these areas, and safety on other equipment, noise, stress, breeding, rack, cage and safety cabinet must be carefully checked. In addition, it is imperative to have equipment and methods to achieve complete disposal of infectious food, feces, urine, bedding and animal carcasses. It is essential to keep certain marks on such areas and admit only authorized personnel including authorized investigators, facility technicians and managers.
Experimental Methods – It is imperative to set up a manual on management methods of animal facilities, which investigators and facility technicians must master. For those experiments on contamination, a manual must be established with reference to Experiment Safety Guidelines of Seoul National University Environmental Safety Institute.
2. Transgenic Animals
The fundamental standard for handling transgenic animals is to manage them in their cages or animal facilities so that they will not escape from there. If a transgenic animal is let loose in wilderness, something unpredictable may occur such as environmental or biological pollution. Thus, careful attention is imperative to prevent them from escaping. In order for such prevention, a wire-gauge must be built on drainage or a ventilating opening, or turning the mouse to an opposite direction when entering or exiting those facilities above. For those animals in which infectious microbe may be produced upon inducting genes, they must be treated in the same manner as those animals in infection experiments.
3. Occupational Health and Safety of Personnel
An occupational health and safety programs must be part of the overall animal care and use program. The program must be consistent with Seoul National University Environmental Safety Institute’s regulations that will depend on the facility, research activities, hazards, and animal species involved.
4. Hazard Identification and Risk Management
Professional staff who conduct and support research programs that involve hazardous biologic, chemical, or physical agents (including ionizing and nonionizing radiation) should be qualified to assess dangers associated with the programs and to select safeguards appropriate to the risks. Potential hazards—such as animal bites, chemical cleaning agents, allergens, and zoonoses—that inherent in or intrinsic to animal use should also be identified and evaluated. Health and safety specialists with knowledge in appropriate disciplines should be involved in the assessment of risks associated with hazardous activities and in the development of procedures to manage such risks. The extent and level of participation of personnel in the occupational health and safety program should be based on the hazards posed by the animals and materials used; on the exposure intensity, duration, and frequency; on the susceptibility of the personnel; and on the history of occupational illness and injury in the particular workplace.
5. Personal Training
Personnel should be trained regarding zoonoses, chemical safety, microbiologic and physical hazards (including those related to radiation and allergies), unusual conditions or agents that might be part of experimental procedures (including the use of genetically engineered animals and the use of human tissue in immunocompromised animals), handling of waste materials, personal hygiene, and other considerations (e.g., precautions to be taken during personnel pregnancy, illness, or decreased immunocompetence) as appropriate to the risk imposed by their workplace.
6. Personal Hygiene
It is essential that all personnel maintain a high standard of personal cleanliness. Clothing suitable for use in the animal facility and laboratories in which animals are used should be supplied and laundered by the institution. Disposable gloves, masks, head covers, coats, coveralls, and shoe covers might be desirable in some circumstances. Personnel should wash their hands and change clothing as often as necessary to maintain personal hygiene. Outer garments worn in the animal rooms should not be permitted to eat, drink, use tobacco products, or apply cosmetics in animal rooms.
1. Guide for the care and use of laboratory animals. ILAR(Instituteof laboratory animal resources) Committee on NRC, 1996, National Academic Press pp 125 (http://www.nap.edu/readingroom/books/labrats/)
2. The animal welfare act. USDA, 1985 (http://www.nal.usda.gov/awic/legislat/usdaleg1.htm)
3. 2000 Report of the AVMA Panel on Euthanasia (http://www.avma.org/resources/euthanasia.pdf)
4. Laboratory animal anesthesia: An introduction for research workers and technicians. PA Flecknell. 1987. Academic press pp156
5. Experiment Safety Guidelines. Seoul National University Environmental Safety Institute. 2003, 3rd ed., Dongwha Books: 307