Instructions for Completion and Submission of an Application for Animal Use in Research

INSTRUCTIONS FOR COMPLETION AND SUBMISSION OF AN APPLICATION FOR A PROTOCOL FOR USE OF ANIMALS IN RESEARCH OR TEACHING

IUPUISCHOOL OF SCIENCE ANIMAL CARE AND USE COMMITTEE

Completion and signing of this form are the responsibility of the principal investigator or faculty member in charge. Completion of the approval process will fulfill Public Health Service and USDA Animal Welfare Act requirements, and will serve to remind users and the public of the IUPUI School of Science's commitment to humane care and use of animals. For proper review and action, complete and submit this application according to the following instructions.

TABLE OF CONTENTS

Page

Application form

Cover page...... 2

Item 1:Lay-oriented summary of purpose...... 3

Item 2:Alternatives, rationale and justification...... 3

Item 3:Lack of duplication...... 3

Item 4:Personnel...... 4

Item 5:Animal husbandry...... 4

Item 6:Disposition and euthanasia...... 5

Item 7:Project description...... 6

Application procedures

Submission of the application...... 12

Review process...... 12

Amendments...... 12

Continuing review...... 12

Suspension of animal use privileges...... 12

Other application forms...... 13

APPLICATION FORM

Copies of the application form, amendment form, and continuing review form are available from the Science Animal Resource Center (SARC) Office.

Cover page: Fill in the name of the principal investigator, his/her rank, department, campus address, and office telephone number. Co-investigators should be listed under Personnel (item 4). Provide a descriptive title of the project. If proposals are submitted for funding consideration under a different title, the title of the project shown on this cover sheet should be listed on the Routing Sheet provided by the IUPUI School of Science Animal Care and Use Committee, (IACUC), which will accompany the route sheet. Give both the anticipated start and completion dates for the project. As mandated by law, you will be required to complete a new application form every 3 years if the project is for more than 3 years.

State the anticipated source of funding for this project. More than one funding source may need to be listed if the project is multifaceted or if the same use of animals applies to more than one project. For external agencies, the name of the agency; for internal funding, state whether the source will be departmental, school, or university.

Complete the checklist of items describing this project. This checklist helps to highlight items that are of potential concern and are required to be reviewed by the IACUC. If you check the box for radioactivity or chemical / biological hazards (because of recombinant DNA, transgenic, knockout, or other genetically altered animal use) you are required to obtain approval for their use from the appropriatecommittee in advance of IACUC approval.

Read the statement regarding your responsibilities in the use of animals in research, and indicate your agreement by signing and dating the completed form.

Item 1: Lay-oriented summary of purpose: Using layman's language, summarize the aims and objectives of this project, including its long-term goals, in a manner that communicates the importance of the project to the scientific community and to the public interest, or the teaching objectives that relate specifically to the use of animals for instruction.

Item 2: Alternatives, rationale and justification: Explain why animals are required for this project. You must perform a computer search (see below) to determine if alternatives to animal use exist for the proposed research. If alternatives (e.g., in vitro methods, computer simulations) exist, justify why these alternative methods were not selected for this project. Justification for any method involving more than momentary pain and distress is also required (see Project Description section for explanation). A computer search must be performed to look for alternatives to the method(s) and if alternatives are found, justify why they cannot be used for this research.

Justify the appropriateness of the species and strain to be used. Explain why a “lower” species cannot be used for this project and why the particular strain has been chosen. If the study involves transgenic, knockout, or other genetically altered animals, approval from the appropriate committee is required prior to IACUC approval. Give a rationale for the number of animals to be used, based on a statistical power analysis, previous findings in the laboratory, or existing literature. Justify the number of animals needed per group, as well as the total number to be used, i.e., also justify the number of groups, replicates, expected attrition rate, etc.

A description of the computer searches for alternatives to both the use of animals and the use of methods involving more than momentary pain and distress must be provided. Include the database(s) searched, the years covered by each search (a period of at least ten years is recommended), keywords (must include “alternatives” as a keyword in both of the searches), and the date each search was performed.

Item 3: Lack of duplication: Replication and extension of previous findings is crucial to the scientific process, but mere duplication or minor modification of other work should be avoided if it reflects insufficient examination of the literature or lacks valid scientific justification. Provide assurance and include an explanation of the basis for concluding that the project does not unnecessarily duplicate previous research. Describe the computer search conducted to determine if similar research has been conducted, include: database(s) searched, years searched (a period of at least ten years is recommended), keywords used, and the date the search was performed. If there is any duplication of prior work, provide a justification for this project. If the search found no pertinent literature, so state.

Item 4: Personnel: List the title (e.g., principal investigator, co-investigator, research associate, assistant, or trainee) and names for each person involved in the project who will have direct live animal contact. Describe their roles in this project (e.g., feeding and weighing, euthanasia, blood sampling, immunizations and inoculations, surgery, anesthesia, postoperative care) and their qualifications to carry out this role. All personnel who will be in contact with either animals or animal tissues must be offered the opportunity to participate in the University's Occupational Health Program.

All persons who have direct contact with live animals are required to attend the annual SARC animal use training sessions. Persons who miss this session or who join the project after the session is given for that academic year must utilize the independent training guide offered by the IACUC. It is the responsibility of the P.I. to insure that all of their personnel receive all relevant training. Documentation of training is necessary before any individual is allowed to use or handle animals.

Item 5: Animal husbandry: Indicate whether the source of animals will be a commercial vendor or another source. If a noncommercial vendor is to be used, you must obtain assurance that the animals are free of viral, bacterial or parasitic disease, and are obtained from facilities that follow Public Health Service animal welfare policies. Identify all noncommercial institutions and facilities including sources within IndianaUniversity (but outside of the IUPUI School of Science).

The total number of animals to be used in this project, the estimated average population and estimated average number of animals housed per cage information is used by the School of Science Space Committee to project space needs. The P.I. is responsible for the use of animals, tracking the number of animals used, and all required record keeping.

Describe any animal care procedures that deviate from usual procedures. Examples include single housing justification, quarantine isolation (e.g., if animals will be received from a noncommercial vendor), reverse or other nonstandard lighting requirements, special caging, and experimental diets. Specify who will be responsible for nonstandard care, e.g., who will give the animals nonstandard diets or where nonstandard caging will be obtained.

Standard animal husbandry followed in the School of Science is described in the Guide for the Care and Use of Laboratory Animals published by the Public Health Service; contact the IACUC or SARC to view a copy of this guide.

Locations housing animals for more than 12 hours outside of the main SARC facility must be inspected and approved prior to housing animals in that setting. State how many animals will be involved, where they will be kept and for how long, and why housing outside the main facility is necessary.

Item 6: Disposition and Euthanasia: State what will be done with the animals after completion of the project. Three possible options are euthanasia (see below), transfer to another active (approved) protocol within the school (either the same or different PI), or shipment outside of the school. IACUC approval is required before animals can be transferred to other protocols or investigators: such a transfer should be stated in this application as well as in the recipient's protocol (a recipient's protocol may be amended to add this source of animals). Collaborators outside of the School of Science must provide documentation that they are approved to receive these animals for research; a copy of their IACUC's approval page is sufficient.

The euthanasia method must minimize pain and distress to the animal. When euthanasia is to be accomplished by lethal injection, specify the drug, dose, and route of administration to be used. It is necessary that following euthanasia by chemical methods, death be assured by physical means, such as the creation of a bilateral pneumothorax, decapitation, severing major blood vessels or organ removal. Methods of euthanasia inconsistent with the recommendation of the American Veterinary Medical Association (AVMA) Panel on Euthanasia must be specifically justified. Contact the IACUC to see the AVMA recommendations.

University policy on Anesthetic Gas Safety requires researchers and staff to complete an on-line training course before using anesthetic gases

List the specific criteria for each species that should be used to determine if animals should be euthanized prior to completion of the study (e.g. severe injury, decreased food intake, percent loss of body weight). Define the humane pre-death end points for the animals on this study.

If euthanasia is an integral part of tissue procurement (e.g., intracardial perfusion), as compared to removing tissues from animals after euthanasia, specify the methods to be used (e.g., lethal injection, physical method, inhalation of anesthetic gases or carbon dioxide). Provide details on drugs, doses, and routes of administration. List who will perform the tissue procurement or surgical procedures.

Item 7: Project description: State your hypothesis and a brief methodology. For the purposes of review, project descriptions fall into five very general categories: 1) all experiments are done on tissues removed after the animals are euthanized; 2) procedures are performed on live animals but neither hazardous materials nor surgery are involved; 3) procedures involving hazardous materials are performed on live animals; 4) surgery; 5) experiments in which animal suffering and death is required as data to provide experimental validity. In this section, you need to give a detailed description of the use of animals but not a long discourse in methodologies unrelated to animal use.

Regardless of the general category into which the project falls, you need to provide the following information:

Species and strain (if applicable)

Age (in some cases, a more general description such as adult or young adult is adequate)

Sex (if either sex may be used, this should be stated)

Numbers of animals to be used in each experiment or teaching exercise

Total number of animals for the entire project
(identify instances where animals are to be assigned to more than one treatment group)

Whether procedures involve pain or distress

A timeline(s) that begins with animal procurement and ends with euthanasia/disposition

The Animal Welfare Act defines a painful procedure as any procedure that would reasonably be expected to cause more than slight or momentary pain or distress in a human being. Expansion of this definition in the Federal Register by the USDA broadens it to include procedures even when the pain is relieved or ameliorated, e.g., even when terminal surgery or tissue extraction is performed on a fully anesthetized animal that is still alive.

University policy on Anesthetic Gas Safety requires researchers and staff to complete an on-line training course before using anesthetic gases

1) If animals are to be euthanized and then serve as a tissue donor, state what materials will be taken from the animals (e.g., muscle, liver, lymphoid tissues) and give a very general statement as to the nature of the experiments to be done. For example, "Proteins will be isolated from liver and analyzed with biochemical techniques to study protein biogenesis." Do not give detailed descriptions of laboratory methods.

Version: June 11, 2009

2) If manipulations are to be performed on live animals, give a detailed description of the procedures and the expected chronological sequence.

Include a different timeline for each group of animals that goes through a unique chronological sequence, i.e., use in multiple procedures versus use in a single procedure, etc. Where multiple procedures are performed on any single animal, please include the expected recovery periods between procedures.

In all cases in which animal pain or distress could be potentially involved, please provide details about the criteria which will be monitored and monitoring frequency that will be employed to limit such distress to the greatest degree possible. Specific criteria must be provided for removal of the animal from the stressful or painful manipulation before irreversible suffering occurs.

In cases where animals will be manipulated in environments other than their home cage, please provide a brief description of the environment, its construction, size, and maintenance. Include how the surfaces to which animals are exposed are sanitized between animals.

The following is a nonexclusive list of typical procedures that may be performed and the type of information that should be provided:

Food and/or water restriction: state the purpose and severity (duration and frequency) of deprivation, total time the schedule will be maintained, how food/water will be provided during the restriction schedule, and how the animals will be monitored to assure that adverse effects or distress do not occur.
Experimental diets: if the animals will be fed diets other than nutritionally complete, veterinary recommended diets, indicate the type of insufficiency or nonstandard composition (e.g., protein deficient, high fat). Describe the potential for compromised health and measures taken to monitor or limit distress or disease.
Exposure to adverse stimuli: describe the stimuli (e.g., electric shock; stimuli evoking strong fear, stress, or defensive responses; motor fatigue; sleep deprivation; intense or abnormal environments such as those involving lighting, sound, temperature, or humidity). Justify the type, intensity and duration of stimulus; the means taken to insure that fatigue, stress, pain, and tissue damage will be minimized or prevented; indicate the availability of escape or avoidance of the aversive stimulation.
Prolonged restraints: describe the purpose, method, duration, and frequency of restraint or confinement. Indicate how often the animal will be observed during the restraint, where (building, room) the restraint will occur, and the adaptation to the device.
Immunization protocols: Indicate the adjuvant, injection sites, volume per site, and frequency of immunization. Indicate the method, frequency, and volume to be used for blood sampling. State what anesthetic will be used, if necessary.
Hybridoma protocols: If animals will be used as hosts for hybridomas, indicate the priming agent and the general nature of the cells to be injected (e.g., rat splenocyte xmouse myeloma NS1). Describe the collection of ascites, including the anticipated number of abdominal taps, the size of needle to be used, and how infections will be monitored and treated.
Chronic injection of drugs or chemicals (other than those listed in category 3, below): Indicate dose, site, frequency, and total time for all treatments. Describe efforts to be taken to minimize or alleviate tissue damage or necrosis.

Version: June 11, 2009

3) Manipulations to be performed on live animals utilizing hazardous materials or conditions must, in addition to answering item #2 above, also describe the hazardous material, its route of administration, dose, or concentration, frequency of administration and duration of exposure. State how the animals will be housed during and after exposure, and whether the hazardous agent will be excreted in urine and/or feces, or exhaled.

Hazardous materials or conditions may include but are not limited to:

Infectious agents

Carcinogens

Toxic chemicals

Recombinant DNA*

Genetically engineered animals*

Transplantable cell lines

Radioisotopes*

Explain the probable animal health effects of exposure. If the animal will be allowed to die as a result of exposure, please provide all of the information requested under part 5 of these instructions (see below).

Describe the specific safety precautions and containment procedures that will be followed in handling animals treated with hazardous agents.

Before initiation of a protocol using radioactive or biohazardous agents, it is the responsibility of the investigator to establish a containment protocol with the SARC in the School of Science and the IUPUI Radiation Safety Office or Environmental Health and Safety Office. In the application for animal use, state whether or not necessary approval from these offices has been obtained or if it is pending. The SARC must be informed of impending use of hazards to ensure the protection of University personnel.

*If the research study involves radioisotopes, recombinant DNA, or genetically altered animals, a copy of the approval from the appropriate committee is required prior to IACUC approval

4) If manipulations are to be performed on live animals, and surgery is included, please insure that all of the following requested information is also included in your description.

Surgery includes the opening of any body cavity, fracture or displacement of any bone, implantation of a subcutaneous device, or disruption of any integumental surface of a living animal by any means other than a hypodermic or biopsy needle.