Azusa Pacific University Institutional Animal Care & Use Committee (IACUC) Animal User Guide

I. Introduction

A. In keeping with federal guidelines, Azusa Pacific University (APU) has established an Institutional Animal Care and Use Committee (IACUC). The purpose of the IACUC is to monitor APUs animal programs, facilities and procedures in order to ensure that these meet current federal, state and local standards. The functions of the IACUC are to:

1. Review, at least once annually, APUs program for humane care and use of animals, and provide a written report of its findings to the office of the Vice Provost at APU. 2. Inspect, at least twice annually, APU’s Animal Care Facility (ACF). 3. Review monthly concerns involving all aspects of the care and use of animals at APU. 4. Make recommendations to the Vice Provost’s office regarding any aspect of the ACF’s facilities or personnel training. 5. Review proposed animal facility construction, remodeling or other alterations in the facility. 6. Review protocols as they relate to the care and use of animals in accordance with the principles specified in this guide. Approval will be recommended, or withheld. If withheld, the Principal Investigator will be asked to make modifications in order to comply with general guidelines. 7. Review proposed significant changes regarding the use of animals in ongoing activities and approve, require modifications in (to secure approval), or withhold approval.

This guide has been prepared to define the policies and procedures that must be observed by investigators and research personnel concerning the treatment and care of animals at APU. To remain current both at APU and in the field of laboratory animal care, these documents are subject to revision. The IACUC will keep a list of animal users in order to notify all concerned of any changes.

B. The Federal Act of 1966, as amended, contains provisions to ensure that animals used in research receive humane care and treatment. The law provides for regulating the transport, purchase, sale, housing, care, handling, and treatment of these animals. The U.S. Department of Agriculture (USDA) is responsible for administration and enforcement of the Animal Welfare Act through the USDA’s Animal and Plant Health Inspection Service (APHIS). In addition to the USDA, the Bureau of Customs and the U.S. Service (PHS) are responsible, under the animal welfare legislation for monitoring the

1 importation of animals. The Bureau of Customs regulations are published in Title 19 of the Code of Federal Regulations; PHS regulations appear in Title 42 of the Code (‘Guide for the Care and Use of Laboratory Animals, 2005, http://www.nap.edu/readingroom/books/labrats ).

The U.S. PHS has an animal welfare policy relating to the use of animals in research and other biomedical activities supported by grants, contracts, and awards from the PHS, which includes the National Institutes of Health (NIH). The PHS requires awarded institutions to conform to the “U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Education,” developed by the Interagency Research Animal Committee (IRAC). In addition to the principles, the PHS and NIH endorse the “GUIDE for the Care and Use of Laboratory Animals,” published by the Institute of Laboratory Animal Resources (ILAR) of the National Research Council (NCR). The requirements of the Animal Welfare Act are set forth under the Regulations and Standards in the Code of Federal Regulations (CFR). The GUIDE is used as a basis for evaluating an institution’s animal care and use program, whether this is done by the institution’s Animal Care and Use Committee, or through accreditation by the American Association for the Accreditation of Laboratory Animal Care (AAALAC, http://www.aaalac.org ); (Animal Welfare; Final Rules; 9 CFR Parts 1 and 2. Federal Register , Vol. 54, No. 168, August 31, 1989, P. 36112-36163, http://www.nal.usda.gov/awic/legislat/awafin.htm ).

The Food and Drug Administration (FDA) has included provisions for adequate housing and care of animals used in research, testing, and for the production of biological products in its regulations: Good Laboratory Practices (GLP, part 58), Good Manufacturing Practices (GMP, part 211), and in subchapter F, part 600, Biological Products (‘Good Laboratory Practice Guidelines, 2005, http://www2.sjsu.edu/faculty/chem55/55glpout.htm ).

California also regulates the acquisition and care of experimental animals. The principal regulator agencies of the state are the Department of Public Health (Veterinary Public Health Unit), the Department of Fish and Game, and the Department of Food and Agriculture (Division of Animal Husbandry). California laws governing the care and use of laboratory animals are set out in the Health and Safety Code, Fish and Game Code and Administrative Code. Generally, state and local laws regulating and handling treatment of laboratory animals meet Federal requirements. All legislation and guidelines require that laboratory animals must be treated humanely; housed in clean, healthful environments; and be fed and watered according their individual needs.

C. The Animal Care Facility (ACF) at APU provides for proper housing and maintenance of research, testing and production of animals. The ACF is in the process of being licensed by the USDA, and will adhere to the regulations set forth in the Animal Welfare Act. In addition, FDA, GLP and GMP regulations

2 are to be followed. The NIH endorsed “GUIDE for the Care and Use of Laboratory Animals” (the GUIDE) and the “U.S. Government Principles for the Utilization and Care of Vertebrate Animals used in Testing, Research and Education” are the standards by which the ACF must operate. Establishment of the IACUC at APU is conditional on meeting federal guidelines. The IACUC uses the GUIDE as a means to evaluate the animal care and use program at APU. It is intended that the IACUC and this Animal User Guide not limit an investigator’s freedom, but encourage all researchers at APU to plan and conduct animal experiments in accordance with scientific and humane principles. Although the ACF and IACUC have been assigned primary responsibility to monitor the animal care and use program of APU, every investigator at APU has a scientific and moral responsibility for observing the laws and regulations governing the treatment and care of laboratory animals (‘Guide for the Care and Use of Laboratory Animals, 2005, http://www.nap.edu/readingroom/books/labrats ).

II. IACUC Appointment & Membership

A. These requirements are found in Title 9 C.F.R., Chapter 1, Subchapter A - Animal Welfare, Parts 1, 2, and 3. The requirement for an IACUC is set forth under the authority of section 13(b) of the Animal Welfare Act (AWA) (7 U.S.C., 2143). The IACUC regulations promulgated under this authority are set forth in Title 9 C.F.R., Chapter 1, Subchapter A - Animal Welfare, Part 2, Section 2.31. Section numbers are given for reference to the actual wording of each requirement (Animal Welfare; Final Rules; 9 CFR Parts 1 and 2. Federal Register , Vol. 54, No. 168, August 31, 1989, P. 36112-36163, http://www.nal.usda.gov/awic/legislat/awafin.htm ):

An IACUC committee shall consist of at least three members, one of whom is the attending of the research facility and one of whom is not affiliated in any way with the facility other than as a member of the committee, however, if the research facility has more than one Doctor of (DVM), another DVM with delegated program responsibility may serve. The research facility shall establish the Committee for the purpose of evaluating the care, treatment, housing, and use of animals, and for certifying compliance with the Act by the research facility:

The requirements, membership, duties, and responsibilities of the IACUC and the Research Facility are as follows:

1. The Vice Provost of each research facility is required, by law, to appoint an IACUC. Those persons appointed to this committee must be qualified through experience and expertise to assess the research facilities animal program, facilities and procedures.

3 2. The members of the IACUC must be appointed by the Vice Provost of the research facility. The IACUC is to consist of no less than three members as follows:

a. A Chairman.

b. A Doctor of Veterinary Medicine (DVM) with training or experience in laboratory animal science and medicine and who has direct or delegated program responsibility for activities involving animals at the research facility. The DVM can not be a researcher who has no direct or delegated responsibility for animals at the facility. Although not specifically prohibited by the regulations, the attending veterinarian should not also be the IACUC chairman.

c. A nonaffiliated member. This person is not to be affiliated with the research facility in any way other than as a member of the IACUC and can not be a member of the immediate family of a person who is affiliated with the research facility. This person is to provide representation for the general community interests in the proper care and treatment of animals. The nonaffiliated member can not be paid for his or her services on the committee. Expenses such as, travel or mileage, meals and parking may be reimbursed however.

d. Should the committee consist of more than three members (NIH requires at least five members on the IACUC), no more than three members can be from the same administrative unit of the facility.

III. IACUC Functions

A. With respect to activities involving animals, the IACUC, as an agent of the research facility shall (http://www.nal.usda.gov/awic/legislat/awabrief.htm ):

1. Review the research facilities program for humane care and use of animals at least once every six months.

2. Inspect all of the research facilities animal facilities, including animal study areas (where animals are housed for more than 12 hours), for compliance with 9 C.F.R., Chapter 1, Subchapter A, at least once every six months. The exceptions that do not require inspection are areas with free living wild animals in their natural habitat.

3. Prepare reports of the evaluations conducted above and submit them to the Vice Provost of the research facility.

B. Provisions

4 1. The IACUC may determine the best means of conducting the above evaluations.

2. No IACUC member wishing to participate in the evaluation may be excluded.

3. The IACUC may use subcommittees composed of at least two IACUC members to conduct the above evaluations and may invite ad hoc consultants to assist them if desired.

4. The IACUC is responsible for the required evaluations and reports even though it may have been done by a subcommittee.

5. The reports must be reviewed and signed by a majority of the IACUC members and must include any minority views.

6. The reports must be updated at least once every six months after the semiannual evaluation.

7. The reports are to be maintained by the research facility and made available to APHIS and any funding Federal agencies for inspection and copying.

8. The reports must contain the following: A description of the nature and extent of the research facilities compliance with regulations.

a. Must specifically identify any departure from compliance with regulations and must state the reasons for each departure. b. Must distinguish significant deficiencies from minor deficiencies. A significant deficiency is one which affects the health, safety, or well-being of the animal(s). c. Must contain a reasonable and specific plan and schedule for correction with corrective dates for each deficiency. d. Failure to correct any significant deficiency within the established deadlines must be reported by the IACUC, in writing within fifteen days and through the Institutional Official, to APHIS and any funding Federal agency. e. Review and investigate concerns or complaints about the care and use of animals at the research facility whether received from the public or research facility personnel. f. Make recommendations to the Institutional Official regarding any aspect of the research facilities animal program, facilities, or personnel training. g. Review, approve, require modifications in, or withhold approval of proposed activities related to the care and use of animals.

5 h. Review, approve, require modifications in, or withhold approval of significant changes regarding the care and use of animals in ongoing activities. i. Be authorized to suspend an activity involving animals if that activity is not being conducted in accordance with the activities approved by the IACUC.

IV. IACUC Review of Activities Involving Animals

A. Conduct a review of the activities related to the care and use of animals to determine if the proposed activities are in accordance with regulations and standards unless acceptable justification for departure is presented in writing. However, field studies are exempt from this requirement. The IACUC must also determine that the proposed activities, or significant changes to activities, meet the following requirements ( http://www.aphis.usda.gov/ac/cfr/9cfr3.html ):

1. Procedures involving animals will avoid or minimize discomfort, distress, and pain to the animals.

2. The Principal Investigator has considered alternatives to procedures that may cause more than momentary or slight pain or distress to the animals and has provided a written justification and description of the methods and sources used to determine that alternatives were not available (ex. data base names, search words, etc.).

3. The Principal Investigator has provided written assurance that the activities do not unnecessarily duplicate previous experiments.

4. Procedures that may cause more than momentary or slight pain or distress to the animals will:

a. Be performed with appropriate pain relieving drugs unless withholding such drugs is scientifically justified, in writing, by the principal investigator and will continue only for the necessary period of time.

b. Require involvement and consultation with the attending veterinarian during planning of the activity.

c. Not include the use of paralytics without anesthesia. This is a firm prohibition and the use of paralytics without anesthesia is not allowed.

d. Any animal that may experience severe or chronic pain or distress, that can not be relieved, will be painlessly euthanized as soon as possible. If such a protocol does not provide for an early

6 termination and euthanasia of the animals then the principal investigator must justify this procedure, in writing, and the IACUC must review and approve such activity.

e. The animals living conditions must be appropriate for their species and must contribute to their health and comfort. The husbandry, nutrition, and non-medical care of the animals is to be directed by the attending veterinarian or other scientist trained and experienced in the proper care, handling, and use of the species being used.

f. Medical care for the animals must be available and provided as necessary by a qualified veterinarian. This includes after hours, weekends, and holidays.

g. All personnel conducting procedures on the species being maintained or used must be appropriately qualified and trained in those procedures.

h. Activities involving surgery must include appropriate provisions for pre- and post-operative care. This care must be in accordance with established veterinary medical and nursing practices (what is currently accepted as the norm in the veterinary profession).

1. All survival surgery must use aseptic procedures (surgical gloves, masks, sterile instruments, and aseptic techniques). 2. Major operative procedures on non rodents are to be conducted only in facilities intended for that purpose and shall be operated and maintained under aseptic conditions. 3. Non major operative procedures and all surgery on rodents do not require a dedicated facility, but must be done under aseptic procedures. 4. Operative procedures conducted at field sites do not require dedicated facilities but must be done using aseptic procedures.

V. IACUC Review and Expedited Review of Activities

A. Prior to any IACUC review, all members of the committee must be provided with a list of proposed activities to be reviewed.

1. Written descriptions of all proposed activities that involve the care and use of animals must be available to all IACUC members.

2. Any IACUC member may require a full committee review of any activity by requesting such.

7 3. If a full committee review is not requested, at least one qualified member can be designated by the chairman to conduct the review. That person shall review the activities and has the authority to approve, require modifications, o r request a full committee review of the activities.

4. If a full committee review is requested for a proposed activity, approval for that activity may be granted only after a review at a convened meeting of a quorum of the IACUC and with approval of a majority of the quorum present.

5. No IACUC member can participate in a review or approval of an activity in which he or she has a conflicting interest, or personal involvement, except to provide information requested by the IACUC.

6. No member who has a conflicting interest in an activity may be part of an IACUC quorum considering that activity.

7. Consultants: The IACUC may invite consultants to assist in the review of complex issues arising during an activity review. The consultant(s) may not approve, withhold approval, or vote with the IACUC unless they are members of the IACUC.

8. Notification: The IACUC is to notify the principal investigator and the research facility, in writing, of its decision to approve, withhold approval, or modify an activity related to the care and use of animals. If the IACUC withholds approval of an activity it shall advise the principal investigator, in writing, as to the reasons for its decision and give the principal investigator an opportunity to respond. The IACUC may reconsider its decision with documentation in the IACUC minutes, in light of the information provided by the principal investigator.

9. Continuing Reviews: The IACUC is to conduct continuing reviews of activities involved with animals at intervals determined by the IACUC but not less than annually.

10. Suspending Activities: The IACUC may suspend an activity if that activity is not being conducted in accordance with the description approved by the Committee. Any decision to suspend an activity must be made at a meeting of a quorum of the IACUC and a suspension vote by a majority of the quorum present.

11. Reporting of Suspensions: If the IACUC suspends an activity involving animals, the Institutional Official, in consultation with the IACUC, shall review the reasons for suspension, take appropriate corrective action, and report that action with a full explanation to Animal

8 and Plant Health Inspection Service (APHIS) and any funding Federal agencies.

12. Final Approval of Activities Involving Animals: Proposed activities and proposed significant changes in ongoing activities previously approved by the IACUC, may be subject to further review and approval by officials of the research facility. However, those officials may not approve an activity involving the care and use of animals if it has not been approved by the IACUC.

VI. Requirements for Proposed Activities or Significant Changes in an Ongoing Activity

A. A proposal to conduct an activity, or to make significant changes in an ongoing activity, involving animals must contain the following.

1. Identification of the species and the approximate number of animals to be used.

2. A rational for involving animals and for the appropriateness of the species and numbers of animals to be used.

3. A complete description of the proposed use of the animals.

4. A description of procedures designed to assure that discomfort, pain, and distress to the animals will be limited to that which is unavoidable for the conduct of scientifically valuable research. This must include provisions for the use of pain relieving drugs, where appropriate, to minimize discomfort and pain to the animals.

VII. Responsibilities of and Procedures for Animal Users

A. All investigators and research personnel using laboratory animals must adhere to federal legislation and guidelines on animal use and handling to ensure continued permission for experimentation. To protect this privilege, investigators work closely with the Animal Care Facility (ACF), which procures and maintains the animals, and with the IACUC, which reviews and monitors animal use at APU. The following section outlines the requirements and guidelines that all animal users at APU must follow in order to meet the standards of humane treatment set forth by the federal government.

B. Based on the guidelines established by the Interagency Research Animal Committee, these principles shall be followed by all investigators when using laboratory animals in planning experimental and production protocols. The IACUC shall use these principles in reviewing APU’s animal care and use program (‘Animal Welfare Act as Amended, 7 USC, 2131-2156, http://www.nal.usda.gov/awic/legislat/awa.htm )

9 1. The transportation, care and use of animals shall be in accordance with the Animal Welfare Act and other applicable federal laws, guidelines and policies. 2. Procedures involving animals shall be designed and performed with due consideration to their relevance to human or animal health, the advancement of knowledge, or the good of society. 3. The animals selected for procedures shall be of an appropriate species and quality and the minimum number required to obtain valid results. Methods such as mathematical models, computer simulation, and in vitro biological systems should be considered. 4. Proper use of animals, including the avoidance or minimization of discomfort, distress and pain, when consistent with sound scientific practices, imperative. Unless the contrary is established, investigators should consider that procedures that cause pain or distress in human beings may cause pain or distress in other animals. 5. Procedures with animals that may cause more than momentary pain or distress shall be performed with appropriate sedation, analgesia, or anesthesia. If anesthesia would defeat the purpose of the research, the investigator must state this in the protocol and the procedure must be directly supervised by the Principal Investigator. Surgical or other painful procedures should not be performed on un-anesthetized animals paralyzed by chemical agents. 6. Animals that would otherwise suffer severe or chronic pain or distress that cannot be relieved shall be painlessly sacrificed at the end of the procedure or, if appropriate, during the procedure. 7. The living conditions of animals should be appropriate for their species and contribute to their health and comfort. The housing, feeding, and care of all the animals used for biomedical purposes must be directed by a veterinarian or other scientist trained and experienced in the proper care, handling and use of the species being maintained or studied. In any case, veterinary care shall be provided as indicated. 8. Investigators and other personnel shall be appropriately qualified, trained and experienced for conducting procedures on living animals. Adequate arrangements shall be made for their in-service training, including the proper and humane care and use of laboratory animals. 9. Methods of euthanasia shall be consistent with methods approved by the American Veterinary Medical Association and the IACUC. Animals that must be sacrificed for purposes of research must be terminated in a humane manner and the method of euthanasia used must result in immediate death. No animal is disposed of until death is certain. 10. Where exceptions are required to the provisions of these principles, the decisions should not rest with the investigator directly concerned, but shall be made with the IACUC. Such exceptions shall not be made solely for the purposes of teaching or demonstrating.

10 C. All research on production proposals throughout APU that entail the use of laboratory animals or their tissues shall be reviewed by the IACUC. The purpose of the review is to ensure the appropriate and humane use of experimental animals, in accordance with the principles described in the preceding section. Any omission or problems regarding animal care and use that result from protocol review will be resolved by discussions between the Principal Investigator and the IACUC. Any unresolved problems, as well as the recommendations of the IACUC, will be forwarded to the Vice Provost Office and a non-partisan review commission shall be appointed to make a final decision.

D. All APU faculty and staff members who will be working with laboratory animals must have a medical examination and a tetanus immunization (if not current) prior to coming into contact with the animals.

E. All faculty and staff members working with laboratory animals, or designing experiments involving the use of laboratory animals will be given a copy of this guide and are required to read it and sign a cover slip stating they have read and understood the guide and will follow the guidelines and principles described in it. The cover slip is kept on file with their Department Chair or Supervisor. The IACUC will be responsible for distribution of this guide to new and current faculty and staff members who are planning, or currently working with, laboratory animals.

F. Significant departures from the requirements for animal care and use are unacceptable and can result in the withdrawal of permission to use animals experimentally. Proper humane care and treatment of laboratory animals benefits all animal research endeavors. Healthier, better adjusted animal models result in more meaningful research findings, which can be more readily replicated, and obtained more economically.

G. Currently, the animal facility consists of a single enclosed, temperature-regulated, animal housing apparatus. The total number of mice that are housed in the facility ranges from 20-40 balb/c mice. Records of the animal receiving and handling procedures are available at the Biology department. Each of the experiments that are being performed require no invasive operating procedures, no injectables, only a behavioral paradigm with successive euthanasia by carbon dioxide or other inhalant anesthetic. Because of the increased interest in animal research, and USDA requirements for externally funded programs, the following procedures (as of December 2005) are being developed by the IACUC committee:

Facility Standard Operating Procedures

1. Animal Facility Procedures 2. Animal Receiving Procedures 3. Cage Changing of Stock and Experimental Animals 4. Animal Handling (mouse, rat)

11 5. Animal Identification (ear tagging) 6. Rodent Breeding of Inbred, Cogenic or Transgenic Lines

General Animal Procedures

1. Anesthesia (inhalation and injection) 2. Bleeding 1. retro-orbital sinus 2. tail vein 3. cardiac puncture 3. Injections 1. intravenous 2. subcutaneous 3. intramuscular 4. intracranial 4. Euthanasia 1. Overdose of inhalant anesthetic 2. Injectable anesthetic 3. Carbon dioxide 4. 5. Decapitation

H. References

United States Department of Agriculture, http://www.usda.gov/wps/portal/usdahome

Animal Welfare; Final Rules; 9 CFR Parts 1 and 2. Federal Register , Vol. 54, No. 168, August 31, 1989, P. 36112-36163, http://www.nal.usda.gov/awic/legislat/awafin.htm

‘Questions and Answers about the Animal Welfare Act and Its Regulations for Biomedical Research Institutions’, http://www.nal.usda.gov/awic/legislat/regsqa.htm

Guide for the Care and Use of Laboratory Animals, 2005, http://www.nap.edu/readingroom/books/labrats .

Animal Welfare Act as Amended (7 USC, 2131-2156), http://www.nal.usda.gov/awic/legislat/awa.htm

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