Policy on Euthanasia of Laboratory Animals

The term euthanasia is derived from the Greek terms eu meaning "good" and thanatos meaning "death". A "good death" would be one that occurs without pain or distress. For the purpose of this policy, euthanasia is the act of inducing humane death in an animal. The Institutional Animal Care and Use Committee (IACUC) uses the 1993 Report of the AVMA Panel on Euthanasia (Report)1 as its guide in reviewing methods of euthanasia in Animal Use Proposals.

Euthanasia techniques should result in rapid unconsciousness followed by cardiac or respiratory arrest and ultimate loss of brain function. In addition, the technique should minimize any stress and anxiety experienced by the animal prior to unconsciousness. Stress may be minimized by technical proficiency and humane handling of the animals to be euthanized.

IACUC Policy

If the animal use requires the euthanasia of the animals, the method of euthanasia must be included in the Animal Use Proposal form.

Personnel performing euthanasia procedures must be adequately trained in the specific method to be used.

Deviations from the recommendations of the Report must be fully justified in the proposal.

Animals subjected to physical methods of euthanasia, e.g., decapitation or cervical dislocation, should be anesthetized or tranquilized prior to euthanasia. Physical methods without prior anesthesia or tranquilization must be scientifically justified in the proposal.

Acceptable Methods:

Inhalant Anesthetics: Euthanasia by overdose of inhalant anesthetics is acceptable for small laboratory animals. In order of preference, halothane, enflurane, isoflurane, methoxyflurane and ether are acceptable for euthanasia of small animals (< 7 kg).
Carbon Dioxide: Carbon dioxide is acceptable for euthanasia of small laboratory animals. Compressed CO2 gas in cylinders is preferable because the inflow to the chamber can be regulated precisely.
Barbituric Acid Derivatives: Barbiturates (sodium pentobarbital, euthanasia solutions containing pentobarbital) may be used for euthanasia. Intravenous injection of barbiturate is the preferred method for euthanasia of dogs, cats, and pigs. Intraperitoneal injection may be used in situations where this approach would cause less distress than intravenous injection, but must be justified in the Animal Use Proposal.
Potassium chloride: Potassium chloride may be used intravenously to stop the heart in deeply anesthetized animals. It may be used only in combination with existing deep anesthesia.
Cervical Dislocation: Cervical dislocation may be used to euthanize birds, mice, and immature rats (< 200 grams). Animals should be anesthetized or tranquilized prior to euthanasia whenever possible. Cervical dislocation will be approved only when scientifically justified by the user and approved by the IACUC.
Decapitation: Decapitation may be used to euthanize rodents. Animals should be anesthetized or tranquilized prior to decapitation (Exception: neonatal rats and mice up to 4 days old may be euthanized without prior anesthesia). Decapitation will be approved only when scientifically justified by the user and approved by the IACUC.

1. Reference: 1993 Report of the AVM Panel on Euthanasia. Jour. Am. Vet. Med. Assn. 1993;202(2):229-249.

Return to IACUC page

Policy on Physical Restraint of Animals

Physical restraint is the use of manual or mechanical means to limit some or all of an animals normal movement for the purpose of examination, collection of samples, drug administration, therapy, or experimental manipulation.

Animals may be physically restrained briefly either manually or with restraint devices. The period of restraint must be the minimum time necessary to perform the procedure. Personnel administering the restraint must be properly trained to handle the species of animal and specific method of restraint. Restraint devices should be suitable in size, design, and operation to minimize discomfort or injury to the animal.

Prolonged restraint of animals is discouraged and must be specifically justified as essential for accomplishing the research objectives in the Animal Use Proposal and approved by the IACUC.

The use of less restrictive systems that do not limit the animals ability to make normal postural adjustments, such as the tether system for non-human primates, should be used when compatible with the protocol objectives.

When restraint devices are used, they should be specifically designed to accomplish research goals that are impossible or impractical to accomplish by other means or to prevent injury to animals or personnel.

The following guidelines apply to the use of restraint devices:

Return to IACUC page

Evaluating Animal Use Proposals with Transgenic or "Knock-out" Animals

Animals with disabilities that are a result of genetic manipulation must be part of an appropriately approved Animal Use Proposal, and the scientific need for producing such animals must be justified in the proposal. The production and maintenance of colonies of such animals must be limited to the extent required by the study. Animals with genetically-induced disabilities which result in pain or distress must receive appropriate treatment, including analgesics, to the fullest extent possible consistent with the experiments being carried out. Animal husbandry and maintenance of these animals will meet or exceed the basic procedures afforded other animals of the same species. Appropriate veterinary care will be provided for conditions which are unrelated to the genetically-induced condition, or the animal may be euthanized.

Return to IACUC page

Policy on Multiple Major Surgical Procedures

Definitions
  1. Major surgery: Major surgery penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic functions (such as laparotomy, thoracotomy, craniotomy, joint replacement, or limb amputation).
  2. Survival surgery: Survival surgery is any surgical procedure in which the animal is allowed to recover from anesthesia.

Policy

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 IACUC.

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 IACUC will pay particular attention to animal well-being through continuing evaluation of outcomes.

Cost savings alone is not adequate reason for performing multiple major survival surgical procedures.

Return to IACUC page




Guidelines for Evaluating Animal Use Proposals with Potential Pain or Distress Associated with Infectious Diseases or Induced Tumors

  1. The investigator must provide information on any expected pain-related illness-inducing or life-threatening effects of an agent.
  2. When infectious agent are administered, the investigator must monitor the animal for signs of acute or chronic pain:
    1. guarding - attempts to protect, move away, or bite;
    2. crying or flinching on movement or palpation;
    3. mutilation - excessive licking, biting, scratching or shaking;
    4. restlessness - pacing, lying down and getting up, shifting weight;
    5. recumbency - reluctance to move or rise for an unusual length of time.
  3. The investigator must monitor for moribund (dying) signs:
    1. impaired ambulation;
    2. muscle atrophy or signs of emaciation;
    3. signs of severe illness including low activity levels, lack of attentive abilities, bleeding, difficulty breathing, signs of central nervous system dysfunction, chronic diarrhea or constipation;
    4. inability to remain upright.
  4. The investigator must justify painful or moribund states in animals.
  5. The investigator must justify death as an endpoint (e.g., LD50).
  6. In cases involving tumor growths, the maximum size tumor allowed is 10% of the body size. When there is obstruction of mobility or of homeostatic mechanisms, (e.g., eating) less tumor growth is tolerated.
  7. If there is an expectancy of pain or distress which cannot be controlled with analgesics or anesthetics, then the investigator must conduct a small pilot study prior to obtaining IACUC approval for a full study. The investigator and IACUC will make a cost:benefit evaluation of the pilot results.


Return to IACUC page

Guidelines for the Use of Freund's Adjuvant in Laboratory Animals

Freund's complete adjuvant, a water in oil emulsion containing killed, dried Mycobacterium butyricum has been widely used in immunology to enhance antigenicity and stimulate an immune response greater than antigen alone. Freund's incomplete adjuvant (water in oil emulsion only) is also frequently used for similar reasons. The improper or unnecessary use of Freund's complete adjuvant may cause severe inflammation, induration and/or necrosis in laboratory animals. The most severe inflammatory responses in animals are seen following multiple injections of the complete adjuvant.

The IACUC has adopted the following guidelines intended to eliminate or minimize animal discomfort associated with the use of this agent in research:

  1. Before using Freund's complete adjuvant, consider the use of the incomplete form or a different adjuvant. (RIBI, trehalose dimycolate)
  2. If Freund's complete adjuvant must be used, it should be limited to the initial immunizing dose. Use of two or more doses of the complete adjuvant is rarely warranted and must be strongly justified with objective data.
  3. The route of injection should be subcutaneous or deep intramuscular rather than intradermal. Intradermal injections may result in skin necrosis and sloughing. The use of footpad injections is normally prohibited.
  4. For subcutaneous administration, the inoculum containing the adjuvant should be divided into fractions so that no more than 0.05 ml is injected per site. Injection sites should be separated from each other widely enough to ensure continued blood supply to adjacent areas of skin and subcutis.
  5. Intramuscular inoculum should be of a volume no greater than 0.5 ml for rabbits (proportionally less for smaller species), administered by a single injection into a deep muscle mass. The deep lumbar muscles of the rabbit are recommended.
  6. Injection sites should be clean and free of debris and contamination which could result in local infection.
  7. The inoculum should be free of extraneous micobial or other participate contamination. Millipore filtration of the antigen before mixing with the adjuvant is recommended when possible.
  8. The oil and water phases should be thoroughly emulsified. Studies suggest that fully emulsified inoculum produces less undesirable reaction at the injection site.
  9. Rabbits should be tranquilized or lightly anesthetized during inoculation to reduce distress. Acepromazine 1.0 mg/kg body weight, subcutaneously or intramuscularly may be used. If the animal vocalizes or struggles during administration of the immunogen, it should be anesthetized. Ketamine 44 mg/kg and xylazine 10 mg/kg im. can be used.


Return to IACUC page

Institutional Animal Care and Use Committee Policy on "Significant Changes" to Animal Use Proposals

The Public Health Service Policy on Humane Care and Use of Laboratory Animals (Policy) and the federal regulations in Animal Welfare (9 CFR Ch. 1) require that the Institutional Animal Care and Use Committee (IACUC) shall, "review and approve, require modifications in (to secure approval), or withhold approval of proposed significant changes regarding the use of animals in ongoing activities...."

Significant Changes include:

Requests to make significant changes to an approved Animal Use Proposal may be made by requesting approval of an Amendment to the protocol. Investigators may submit requests in letter form to the committee or may submit amended pages of the Animal Use Proposal form.

Requests for amendments must contain:

  1. Title and assigned number of the approved Animal Use Proposal;
  2. Description of the proposed amendment including complete description of any change in invasive procedures, surgery, anesthetic or analgesic drugs, or euthanasia;
  3. Justification for the proposed change(s).

Copies of proposed amendments will be made available to IACUC members in the same manner as new Animal Use Proposals, and will be reviewed at the IACUC meeting.

Return to IACUC page




Use of Di-ethyl Ether as an Animal Anesthetic

  1. Di-ethyl ether (ether) is highly flammable and explosive. It is a hazardous substance which requires careful attention to safety precaution when it must be used. You are encouraged to use it only when no reasonable alternatives exist. When its use is necessary, ether must be used only in facilities designed and approved for its use and in a manner which will avoid the known hazards.
  2. Facilities where ether is used must be designed such that sparks or open flame cannot occur to ignite the material.
  3. Specifically, the use of ether in animal rooms and Animal Resource Facilities procedures rooms is prohibited. There are no Animal Resource Facilities designed for the safe use and storage of ether.
  4. Euthanasia of animals with ether creates a special hazard. Normally disposal of research animal carcasses is via incineration. Tissues of animals killed with ether are saturated with ether and cannot be safely placed in an incinerator.
  5. Alternatives to the use or ether as an animal anesthetic include the gases halothane and methoxyflurane; the injectable barbiturates; and combinations of ketamine with xylazine, acepromazine, and diazepam. In the case of euthanasia, the inhalant anesthetics and barbiturates given in excess are acceptable. For small rodents and lagomorphs, carbon dioxide gas is recommended. For further information, contact Animal Resource Facilities, 777-8106.
  6. With storage of open containers of ether there is a likelihood of the production of explosive peroxides. Therefore, storage of ether after the manufacturers seal on the container has been broken is not recommended. If storage is required, it must be only in explosion proof areas. Storage in Animal Resource Facilities is prohibited.
  7. For safe disposal of unused ether, opened ether cans or out-dated unopened ether containers you should contact Environmental Health and Safety, 777-5269.


Return to IACUC page

Policy for Importation of Rodents

Genetically manipulated animals provide a valuable research tool. Hundreds of different transgenic and gene knockout rodents are now available to investigators. However, most of these very specialized animals are available only from limited sources, usually other research institutions and universities. The procedures for housing and maintaining animals and monitoring their health status vary widely from institution to institution. The risks of importing infectious disease agents and internal and external parasites are increased, placing our established non-infected colonies at great risk.

To reduce the opportunities for introducing potentially devastating agents into U.S.C. animal facilities, the Institutional Animal Care and Use Committee (IACUC) has established the following policy for importation of rodents from other institutions.

  1. Investigators requesting to introduce animals must have an Animal Use Proposal approved by the IACUC prior to making the request for animals.
  2. No animals will be allowed into the U.S.C. facilities until the health status of the origin colony has been reviewed by the Attending Veterinarian. The review may include submission of whole animals and/or specimens to a reference diagnostic laboratory. All incoming shipments of animals will be free from adventitious viruses and parasites. Diagnostic charges incurred will be charged to the investigator's account.
  3. All shipments of animals to U.S.C. will be coordinated through Animal Resource Facilities (ARF).
  4. Imported animals will be placed in quarantine in an ARF approved facility until such time as they are deemed acceptable for introduction into the general animal housing area.
  5. Quarantined animals may be removed for euthanasia and harvest of tissues. No animal removed from the quarantine facility will be allowed back into any animal holding areas. Quarantined animals may not be housed in investigators' laboratories.


Return to IACUC page

IACUC Policy on Nonhuman Primates

Nonhuman primates have been and will continue to be very valuable animal models in biomedical research. For some types of research viable alternatives to the use of nonhuman primates are not available. It is clear, however, that the use of nonhuman primates presents many serious problems for animal health and well being, management of the animals, and health risks to the researchers and animal care staff. The use of nonhuman primates, especially Old World primates requires specialized housing and facilities, intense, continual training of staff, and well-organized support staff providing medical surveillance and treatment, hazard assessment, and security. The animal facilities at U.S.C. do not routinely include housing and programs for use of nonhuman primates.

The IACUC strongly encourages investigators who require nonhuman primates for their research to conduct the animal work in specialized primate facilities such as those in the National Institutes of Health Regional Primate Research Centers or university primate research centers. The IACUC and the University Veterinarian will assist the investigator with locating and evaluating the sites, reviewing the animal use programs, and developing protocols and contracts.

If the research cannot be conducted at an off campus site, the investigator is responsible for coordinating his needs with the IACUC and Animal Resource Facilities. The investigator is responsible for assuring that funding for specialized caging, necessary training of personnel, occupational health surveillance of personnel, and other costs associated with establishing a primate colony are available. The IACUC will assist the investigator and may recommend suitable consultants for developing programs and procedures.

No Nonhuman primates will be brought into U.S.C. animal facilities until a complete program for primate care and use is in place and has been coordinated with all the involved University elements including Health and Safety Programs, University Specialty Clinics, U.S.C. Police, and Media Relations.

Return to IACUC page




Reporting Concerns About the Use of Laboratory Animals

The following information is provided to University of South Carolina investigators, technicians, and students who work with or handle laboratory animals. University Policy ACAF 5.04, "Protocol for Reporting Misuse of Animals" applies to laboratory animals used in teaching or research.

The University recognizes that the use of animals for teaching and research is fundamental to advances in biology and medicine. The momentum of discovery in the medical, biological and behavioral sciences has steadily increased and the application of this knowledge has brought incalculable benefits to society. Laboratory animals have played an indispensable role in these advances, and for the foreseeable future will continue to be essential to the success of our research and training programs.

The University accepts an ethical and scientific responsibility to provide humanely for the welfare of all animals used in education and research at USC. The University recognizes that the public has a concern for the welfare and well-being of the animals. The Institutional Animal Care and Use Committee (IACUC) has oversight over the care and use of animals at the University.

Under the Animal Welfare Act, the IACUC is given the function of reviewing and investigating concerns involving animal care resulting from complaints by University personnel or the public. The IACUC is obligated to assess the validity of the concerns and to respond in an appropriate manner to claims of disregard for the welfare of laboratory animals. Any misuse of laboratory animals whether intentional or unwitting, will be addressed.

The IACUC is given responsibility for handling charges of animal misuse in an expedient, just, and appropriate manner, with regard for the protection of both the person(s) being accused and the person(s) submitting the allegations.

Reporting Procedure

Any faculty member, student, staff member, or other individual who feels that animal misuse may have occurred has an obligation to report their concern to the veterinarian in charge of USC’s animal facilities. Alternatively, reports should be made to the chairman or some other member of the USC IACUC. Concerns may be reported in writing and signed (preferred by IACUC) or by an anonymous tip. Persons may informally consult with the veterinarian, chairman or a committee member prior to initiation of a formal report.

The Office of Research Integrity (ORI), Department of Health and Human Services (DHHS), has issued guidance on protection of persons reporting animal misuse. The USC IACUC will protect any person that makes good faith allegations of animal misuse to appropriate institutional authorities. In addition, the Public Health Service (PHS) Act Part 493(e) mandates “the protection of informers for (1) good faith allegations of an inadequate institutional response to scientific misconduct allegations and (2) good faith cooperation with investigations of such allegations”. Once an allegation has been made, the USC IACUC will act in a manner which protects the reputation and well-being of both the accused and the accuser.

Persons making wrongful allegations of animal misuse with malicious or vengeful intent, or persons taking wrongful action against the accuser because they have reported suspected misuse are subject to disciplinary actions by the University.

Investigation of Complaints

The IACUC Chairperson will appoint an ad hoc investigating committee of at least two IACUC members. The investigating committee will conduct a review to determine if inappropriate handling or use of animals has occurred and, if so, the extent of the alleged animal misuse.

The investigating committee may review records and interview persons with relevant information including the person accused of inappropriate use, the accuser, and others. The committee may be assisted by outside consultants if it deems them necessary.

Following its investigation, the committee will report its findings to the full IACUC. The IACUC will review the report and make its recommendations. In cases where inappropriate animal handling or use have been determined to occur, the full IACUC will recommend appropriate corrective actions which may include, additional training of personnel, increased oversight, or suspension of further acquisition and use of animals. The committee may take measures to eliminate obvious animal suffering immediately and suspend further animal use.

The IACUC Chairperson will forward the investigation report and the IACUC recommendations to the Institutional Official for further action.

Return to IACUC page




University of South Carolina Policies and Procedures

SECTION       Academic Affairs
NUMBER       ACAF 5.04
SUBJECT       Protocol for Reporting Misuse of Animals
DATE               February 28, 1992

  1. Policy
    1. Under the Animal Welfare Act, the Institutional Animal Care and Use Committee (IACUC) is given the function of reviewing and investigating concerns involving animal care resulting from complaints by University personnel or the public. The IACUC is obligated to assess the validity of and to respond in an appropriate manner to claims of disregard for the welfare of laboratory animals. Any misuse of laboratory animals shall be addressed, whether it is intentional or unwitting.

    2. Charges of animal misuse can damage an individual's career, seriously hamper research progress, and create volatile public reaction. Therefore, charges must be handled in a professional and confidential manner. The least number of persons necessary should be involved at any stage of the procedure. Any inquiry or investigation must be handled promptly and expeditiously and with full attention to the rights of all individuals involved. Persons conducting the investigation must possess the special knowledge necessary to judge the situation but must also have no immediate personal interest in the case.

    3. The IACUC is given responsibility for handling charges of animal misuse in an expedient, just, and appropriate manner, with regard for the protection of both the person(s) being accused and the person(s) submitting the allegations.

    4. Persons making wrongful allegations of animal misuse with malicious or vengeful intent, or persons taking wrongful action against the accuser because of such allegations, are SUBJECT TO DISCIPLINARY ACTIONS BY THE UNIVERSITY OF SOUTH CAROLINA.

      5.
  2. Procedure

    These procedures are designed to recognize the difficulty in determining the extent of animal misuse. The process of inquiry and investigation should be sufficiently flexible to be terminated when it becomes clear that charges are unjustified or that the issue can be resolved appropriately by other means.
    1. Reporting Procedure

      1. Any faculty member, student, staff member or other individual who suspects that animal misuse has occurred has an obligation to report, in writing, that suspicion to the veterinarian in charge of USC's animal facilities. Alternatively, reports should be made to the chairman or some other member of the USC IACUC.
      2. The identity of the person bringing the matter to the attention of the committee will not necessarily be concealed, since this person may be an important witness in an investigation.
      3. Once an allegation has been made, the USC IACUC should act in a manner which protects the reputation and well-being of both the accused and the accuser.

    2. Investigation

      1. An investigation committee will be established by the IACUC chairman consisting of the IACUC chairman or his/her designee, the USC veterinarian, and 1 IACUC member unaffiliated with the University. The investigation committee will make a preliminary review to a) exclude frivolous accusations and b) to determine the extent of alleged animal misuse.
      2. The person accused of animal misuse must be given an opportunity to answer the charges at this time.
      3. If necessary, appropriate measures should be instituted to eliminate immediately any ongoing animal misuse.
      4. Any other persons with relevant information should be interviewed at this time.
      5. If the IACUC investigation committee decides that there are no grounds for a charge of animal misuse and no further inquiry is necessary, a written report should be filed with the IACUC. A copy of the report will be kept in a permanent, confidential file in the Office of the Associate Vice Provost for Sponsored Programs and Research. The accuser and the accused will be notified of the outcome of the investigation. The case is then considered closed.
      6. If the IACUC investigation committee determines that animal misuse has occurred, a report to the full IACUC committee is mandatory.
      7. Investigation of animal misuse allegations should be completed within 30 calendar days of the initial allegation.
      8. In unusual circumstances where additional time is required to review the issues involved, the investigation committee shall keep a written record of the cause for delay. Any delay beyond 60 calendar days shall be reported to the Associate Vice Provost for Sponsored Programs and Research.

    3. Action by IACUC

      1. In cases where animal misuse has occurred, the full IACUC committee will consider the report of the investigation committee to determine appropriate action. Consideration will be given to the following actions by IACUC:
        1. Require appropriate training and corrective action prior to continued use of laboratory animals by the persons involved in animal misuse.
        2. Halt all further acquisition and use of laboratory animals by persons involved in animal misuse.
        3. Notification of the Associate Vice Provost for Sponsored Programs and Research when sponsoring agency funds are involved. This office will file additional reports to relevant sponsoring agencies as required.
        4. If misuse is judged as willful and serious and where the intent of the misuse warrants, referral of case for consideration by the USC scientific misconduct process.
        5. Rejection of investigation committee's determination that animal misuse has occurred.
      2. A final report of committee action in each case will be kept in a permanent, confidential file with the IACUC and in the Office of the Associate Vice Provost for Sponsored Programs and Research. The accused and the accuser will be notified of the final IACUC decision.


Return to IACUC page

The Use of Expired Material and Non-Pharmaceutical Grade Drugs


The use of expired medical materials such as drugs, fluids, or sutures on vertebrate animals is not considered to be acceptable veterinary practice and does not constitute adequate veterinary care as required by the regulations (Animal Welfare Act). All expired medical materials must either be disposed of or segregated into an appropriately labeled, physically separate location from non-expired medical materials.

For acute terminal procedures, expired medical materials may be used if their use does not adversely affect the animal's well-being or compromise the validity of the scientific study. Proper anesthesia, analgesia, and euthanasia are required for all such procedures. Drugs administered to relieve pain or distress and emergency drugs must not be used beyond their expiration date.

Pharmaceutical-grade medications are to be used whenever they are available, even in acute procedures. Non-pharmaceutical-grade chemical compounds should only be used in vertebrate animals after specific review and approval by the IACUC for reasons such as scientific necessity or non-availability of an acceptable veterinary or human pharmaceutical-grade product. Cost savings alone are not an adequate justification for using non-pharmaceutical-grade compounds in vertebrate animals.

Return to IACUC page



Calibration and Inspection of Anesthetic Systems


An anesthetic system is defined as the whole anesthesia delivery apparatus including but not limited to the vaporizer, gas monitors, tubing and F/AIR charcoal canisters. All anesthetic systems used for animal survival surgical procedures must be registered with the Institutional Animal Care and Use Committee and Animal Resource Facility (ARF). Anesthetic systems must be inspected and calibrated once every three years.

The F/AIR charcoal canister (anesthesia gas filter) is specifically designed to remove waste anesthetic gases such as Isoflurane and Enflurane from the operating environment. These canisters have a finite usable life span and investigators must be aware of the total use of a canister to assure that it is still functional.

The manufacturer states that a single F/AIR canister can remove 50 grams of halogenated anesthetic waste. The canisters should be checked before and after each use to determine if it is still functional. The use should be recorded on the canister in the space provided.

Procedure:

  1. Weigh the canister when it is initially put into use and record the weight in the table provided.
  2. After each use, reweigh the canister and record the weight (it will be higher than the start weight).
  3. The canister should be replaced when the weight has increased by 50 grams.

Return to IACUC page



IACUC Guideline on Endpoints in Animal Use Proposals


Moribund /MOR-uh-bund/,adjective: 1. In a dying state; dying; at the point of death.

morbid /Mor-bid/, adjective: pertaining to, affected with, or inducing disease; diseased.

Experimental studies may involve procedures that cause clinical symptoms or morbidity in animals. Ideally, studies should be terminated when the animals begin to exhibit clinical signs of disease (morbidity) if these endpoints are compatible with meeting the research objectives. Such endpoints minimize pain or distress and are preferable to death or moribundity as endpoints which can involve considerably more pain or distress.

Although infrequent, allowing animals to become moribund or die as endpoints may be necessary for some research projects. The moribund condition is defined as a clinically irreversible condition leading inevitably to death. In these studies, animals are permitted to die or become moribund as a result of experimental procedures, in some cases without the use of pain relieving measures, because such measures may compromise the integrity of the study.

It is the policy of the Institutional Animal Care and Use Committee (IACUC) that studies requiring moribundity or death as the endpoint will not be approved without specific written scientific justification. Animals that become moribund will be humanely killed by approved methods.

In those cases where death is the only acceptable endpoint, the investigator must provide an explicit explanation for this endpoint. Particular attention should be paid to providing the rationale for not humanely killing moribund animals, e.g., what additional scientific information will be gained.


Animal Study Proposals utilizing death or moribund condition as an endpoint should contain the following information:

  1. The scientific rational for death or moribund condition as an endpoint, including:
    1. What alternatives were considered, why morbidity as an endpoint cannot be used, and how alternatives will be used whenever possible.
    2. Why pain relieving measures cannot be utilized.
    3. Number of animals to be used and why this is the minimal number of animals required.
    4. Where animals will be euthanized when moribund and if not, what information is to be gained in the interval between moribundity and death.

  2. A plan for the following animal care and monitoring procedures:
    1. Animals involved in experiments that may lead to a moribund condition or death will be monitored daily by personnel experienced in recognizing signs of morbidity (illness, injury, or abnormal behavior) for at least the following: abnormal posture, rough hair coat, head tucked into abdomen, exudate around eyes and /or nose, skin lesions, or abnormal breathing, difficulty with ambulation, decreased food or water intake, or self mutilation.
    2. The frequency of observation will be increased (including on weekends and holidays) when animals exhibit the above or other signs of moribundity. Designated personnel, including a veterinarian, should be notified as soon as animals show signs of disease. An assessment of the animals' condition should be made as soon as possible and a plan of action established.
    3. Consideration will be given to moving animals to individual cages when their condition deteriorates to the point that injury from other animals is likely. Dead animals must be promptly removed.
    4. Written records will be kept of monitoring.


Recognizing Signs of Morbidity and Moribundity

The following are signs and symptoms for judging morbidity (disease/illness) and moribund condition (state of dying) in animals:

Morbidity:

  1. Rapid breathing rate
  2. Breathing rate very slow, shallow, and labored (preceded by rapid breathing)
  3. Rapid weight loss
  4. Ruffled fur (rough hair coat)
  5. Hunched posture
  6. Body temperature less than 30° C (hypothermia) or
  7. Hyperthermia
  8. Ulcerative dermatitis or infected tumors
  9. Anorexia (loss of appetite)
  10. Diarrhea or constipation

Moribund Condition:

  1. Impaired ambulation (unable to reach food or water easily)
  2. Evidence of muscle atrophy or other signs of emaciation (body weight is not always appropriate)
  3. Any obvious severe illness including such signs as lethargy (drowsiness, aversion to activity, lack of physical or mental alertness), anorexia (loss of appetite, especially when prolonged), bleeding, difficulty breathing, CNS disturbance, or chronic diarrhea
  4. Inability to remain upright
  5. Loss of consciousness
  6. Drop in body temperature below 28° C for a prolonged period (6 hours)



Return to IACUC page



Policy and Guidelines on Food and Water Restriction or Deprivation


Definitions:

Standard Food Intake: Animals should be fed palatable, noncontaminated, and nutritionally adequate food daily or according to their particular requirements (Guide, page 38).

Standard Water Intake: Ordinarily, animals should have access to potable, uncontaminated drinking water according to their particular requirements (Guide, page 40).

Food or Water Restriction: Any deviation from the standard food or water intake.

Food Deprivation: Withholding food for longer than 24 hours for simple stomach animals, or longer than 48 hours for ruminants.

Water Deprivation: Withholding water for longer than 12 hours.


IACUC Policy and Guidelines

Food or water restriction/deprivation involving laboratory animals, other than presurgical fasting, are often necessary for behavioral testing. Because these procedures may cause more than momentary or slight distress to the animals, the USC IACUC has established the following policy and guidelines:


  1. Dietary or water restriction/deprivation must be scientifically justified in the Animal Use Proposal (AUP) and approved by the IACUC.
  2. The least restriction/deprivation that will achieve the scientific objective should be used.
  3. 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.
  4. A monitoring program for these animals must be described in the AUP. This should include physiological and behavioral parameters for assessment of pain or distress, including criteria (such as weight loss or hydration state) for temporary or permanent removal of the animals from the experiment. Body weight must not be allowed to drop below 80% of normal (taking into account the normal anticipated growth for that animal).

Return to IACUC page



NIH Policy on Allowable Costs for Grant Activities Involving Animals


Notice Number: NOT-OD-07-044

Release Date: January 26, 2007

Issued by
National Institutes of Health (NIH), http://www.nih.gov

The purpose of this Notice is to clarify that no costs for activities with live vertebrate animals may be charged to NIH if there is not a valid Animal Welfare Assurance and Institutional Animal Care and Use Committee (IACUC) approval. This notice is applicable to grants and cooperative agreements involving activities with live vertebrate animals.


Background

Terms and conditions applicable to all grant awards that involve live, vertebrate animals - including research, research training, experimentation, biological testing, custom antibody preparation, or related purposes - require a valid Animal Welfare Assurance (Domestic, Foreign, or Inter-institutional Assurance, as applicable) approved by the NIH Office of Laboratory Animal Welfare (OLAW), and valid IACUC approval. IACUC approval must be dated within the last three years in order to be valid. IACUCs are not authorized to administratively extend approval beyond three years. Foreign grantees receiving direct support are not required to provide IACUC approval, but must have a valid Foreign Assurance on file with OLAW (see http://grants.nih.gov/grants/olaw/assurance/500index.htm for list of foreign institutions with approved Assurances).


Policy

The Office of Management and Budget Cost Principles and the NIH Grants Policy Statement (NIHGPS) do not permit charges to grant awards for the conduct of animal activities during periods of time that the terms and conditions of the NIHGPS are not upheld. Specific situations under which charges are not allowable are:

  1. The conduct of animal activities in the absence of a valid Assurance on file with OLAW.

    2. USC has a valid Institutional Assurance on file with OLAW

  2. The conduct of animal activities in the absence of valid IACUC approval of the activity. Absence of IACUC approval includes failure to obtain IACUC approval, expiration, or suspension of IACUC approval.

    3. In order to charge animal care costs to an NIH grant, USC investigators must have an approved and valid Animal Use Proposal (AUP). In the absence of a valid AUP, these costs must be paid with other institutional funds.

    In order to meet the above federal requirements, Animal Resource Facilities (ARF) will contact the Principal Investigator (PI) and the corresponding departmental Business Manager to procure an appropriate non-NIH fund number when:

    1. A PI does not have IACUC approval to use animals, but animals are being maintained by ARF and charged to an NIH grant account.
    2. The PI allows an approved AUP to expire, but intends to maintain the animals until an AUP is approved.
    3. The activities on a previously approved AUP have been suspended by the IACUC.

It will be the responsibility of the PI and the department to ensure that per diem charges will be covered by non-NIH funding. If this can not be accomplished, the animals will not be allowed to remain in a University of South Carolina animal facility. The PI and the department will be responsible for notifying Contract and Grant Accounting on all matters related to these requirements.

Questions related to this policy and USC procedures should be directed to Elizabeth Thames, IACUC Administrator, at 7-8564.

Return to IACUC page



Policy of Use of Cell Lines in Live Animals


The confounding effects of viral infections (often without signs of clinical disease) of experimental animals, e.g. mouse hepatitis and ectromelia are well documented in the literature. 1 These infections can have devastating effects on experimental results as well as on production of animals from breeding colonies.2 Of equal concern is the potential for infections among laboratory workers. Infections with zoonotic agents such as hantavirus3 and lymphocytic choriomeningitis4 have been traced to contaminated biological material transplanted into animals.

Biological material and animal products such as cell lines, tissues, and tumors have been repeatedly incriminated as vehicles for the introduction of animal pathogens into animal colonies.

The prevention of accidental introduction of viral infections into research animals is essential to maintenance of the animal colonies at the university. To maintain the integrity of the animal colonies the Institutional Animal Care and Use Committee has established the following guidelines regarding the introduction of cells, tumors, and other biologic products into experimental animals:

  1. Investigators are responsible for ensuring that the biologic materials used in their study will not endanger the health of study animals or other animals housed in the facility. Investigators planning to introduce tumors, tissues, or cells into experimental animals must complete the relevant section of the Animal Use Proposal (Section IX).
  2. Cell lines, tumors, or other biologic materials originating from rodents or passaged through rodents must be certified free of murine pathogens prior to their introduction into USC animal facilities. This includes cell lines purchased from ATCC or other commercial sources.
  3. Investigators must provide the IACUC with documentation that the material has been tested and verified free of pathogens before introducing these materials into animals. Cell lines, tumors or other materials for which no documentation is provided must be tested. The preferred method for testing material is by polymerase chain reaction (PCR) procedures.
  4. Any material found to be positive for murine pathogens must not be used. Once the material is shown to be cleared of the agent(s) and the proper documentation is on file in the ARF office, the material may be used for experiments.
  5. Human cell lines that have not been passaged through rodents do not require testing for murine viruses.
  6. Cell lines passaged through rodents and frozen for storage and later use must be retested when thawed. More frequent testing of cells lines may be required if: 1) rodent pathogens are detected in the room where the cell lines are being used; 2) the material will be transferred to animals in a campus facility other than that in which it was originally used; 3) cell lines have been passaged through animals housed in an off-campus facility and returned to USC; or 4) as otherwise determined by the Attending Veterinarian and/or IACUC.



1 Committee on Infectious Diseases of Mice and Rats, Institute of Laboratory Animal Resources. Infectious Diseases of Mice and Rats Pub 397 (National Academy Press, Washington, DC, 1991.
2 Baker, D.G. Natural pathogens of laboratory mice, rats, and rabbits and their effects of research. Clin. Microbiol. Rev. 11, 231-266 (1998).
3 Lloyd, G. & Jones, N. Infection of laboratory workers with hantavirus acquired from immunocytomas propagated in laboratory rats. J. Infect. 12, 117-125 (1986).
4 Hinman, A.R. et al. Outbreak of lymphocytic choriomeningitis infections in medical center personnel. Am. J. Epidemiol. 101, 103-110 (1975).

Return to IACUC page



Training of Personnel for Laboratory Animal Use or Care


The University of South Carolina recognizes that the use of laboratory animals for teaching and research is fundamental to advances in biology and medicine. The use of laboratory animals is subject to a multitude laws, policies, regulations, and standards. Two such laws/policies include the Animal Welfare Act (AWA) and the Public Health Services (PHS) policy, which require documentation that personnel are appropriately trained in animal care and use. To comply with these regulations, the University of South Carolina utilizes two primary types of personnel training: 1) Laboratory Animal Training Association (LATA) Online Training, and 2) Hands-on Rodent Workshop. All persons using or caring for animals in research, testing, or education at the University of South Carolina must complete the specified training described below.

  1. Required Training:

    1. All personnel working with laboratory animals are required to complete the following modules:
      1. The Humane Care and Use of Laboratory Animals module.
      2. The specific species module for each species listed in the protocol.

    2. All persons performing surgery on laboratory animals are required to completed the following modules:
      1. Aseptic Surgery of Rodents
      2. Anesthesia and Analgesia of Rodents

  2. Optional Training:

    Animal Resource Facilities provide hands on training for individuals working with mice and/or rats. These workshops cover the basics of rodent research and techniques can be tailored to meet the needs of the individuals receiving the training. Participation is not required, but strongly encouraged.


Return to IACUC page