Protocol Application for Animal Use Instructions General Information
The Institutional Animal Care and Use Committee Protocol Application for Animal Use (the Application) must be completed by the Principal Investigator and approved by the Institutional Animal Care and Use Committee (IACUC) prior to beginning a project that involves live vertebrate animal use, as well as for significant revisions to an existing protocol or renewal of an expiring protocol. Animals may not be ordered or used in experiments, teaching or breeding without an approved protocol.
The most recent version of the Application can be obtained from the Creighton University IACUC website:
http://www.creighton.edu/fileadmin/user/ResearchCompliance/IACUC/forms/IACUC_Application_-03-16-09_-_For_posting.doc A good overview of the process of writing a grant application that involves the use of animals can be found on NIH's Office of Laboratory Animal Welfare website, http://www.niaid.nih.gov/ncn/clinical/researchanimals/tutorial/index.htm. In particular, the section Write your protocol (http://www.niaid.nih.gov/ncn/clinical/researchanimals/tutorial/write_your_protocol.htm) succinctly hits the main points of preparing your application for the IACUC. The underlying principles that the IACUC follows in evaluating your application are expressed in the U.S. Interagency Research Animal Committee's Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and Training, on pages 4-5 of the Public Health Service Policy on Humane Care and Use of Laboratory Animals (PHS Policy). The Guide for the Care and Use of Laboratory Animals (The Guide) provides more detail. Both publications are found on the IACUC website. http://www.creighton.edu/fileadmin/user/ResearchCompliance/IACUC/
The Health Science Reference Librarian (280-5138 or email@example.com) can help you construct searches to optimize the choice of animal and species (question B5), prevent duplication (question B5), and minimize pain and distress (question E9). These searches should be conducted early in the development of the experimental design so that alternative approaches (see Animal Welfare Act) can be incorporated into the design.
Animal Welfare Act regulations require Principal Investigators to consider alternatives to procedures that potentially cause more than momentary or slight pain or distress to the animals. Whenever possible, you should Replace, Reduce, and Refine (the three Rs) potentially painful procedures. This means:
Replacing animals in part or full with non-animal systems (for example, in vitro, computer or mathematical models) or less sentient animal species (for example, insect or molluscan models instead of mammalian models).
Reducing the number of animals to the minimum required to obtain scientifically valid data through a priori consideration of appropriate experimental and statistical design (for example, more advanced statistical tests, power analyses).
Refining procedures to lessen or eliminate pain or distress, thereby enhancing animal well-being (for example, more effective analgesia and anesthesia, less-invasive surgical techniques, terminating experiments at earlier stages of morbidity).
The Attending Veterinarian can provide advice on refining procedures, choosing drugs and dosages, etc. The IACUC Chair, IACUC Animal Research Specialist, or other IACUC members may also be consulted for assistance in preparing the Application.
Applications ideally encompass a single complete set of experiments testing closely related hypotheses examined with similar procedures. In most cases, you should submit only one Application for any set of experiments, even if you will seek funding from several sources. Multiple protocols may be appropriate when a large breeding colony is involved to provide animals for several sets of experiments; in this case, you may find it more convenient to submit one protocol for breeding work and others for the experimental work. If the requirements of a grant include verification that the animal work has been approved by the IACUC, it is advantageous that all the work be covered by a single protocol. The title of the protocol does not have to match that of the grant proposal.
Answer all questions on the Application; answer N/A where appropriate rather than leaving an answer blank. Please use language suitable for a lay reader throughout the application so that all members of the IACUC can understand your protocol. Federal regulations require that the IACUC include at least one non-scientist. If the non-scientist cannot readily understand the Application, it will be returned for clarification and resubmission.
The Application is designed to be completed in your word processing program. Check boxes can be marked within Microsoft Word by double-clicking on the box and then selecting “checked” under “default value.” The tables included in the Application can be expanded or duplicated as needed by copying and pasting, or deleted when not applicable. Handwritten Applications will not be accepted.
All personnel working with animals must abide by the Animal Resource Facility (ARF) Standard Operating Procedures, found online at http://www.creighton.edu/fileadmin/user/ResearchCompliance/IACUC/ Investigators planning to use animals with special needs or obtained from sources other than approved commercial vendors should contact the ARF Manager prior to submitting an Application to ensure that housing and care can be provided. Animals from non-approved facilities must be quarantined; please design your experimental timelines to account for the quarantine period.
All Applications are initially subject to full committee review at a full committee meeting. The IACUC meets on either the last or the next to the last Monday of the month. The IACUC does not conduct less than full committee review of Applications.
The original signed Application and an electronic version, must be submitted to the IACUC Office no later than 5:00 p.m. of the first Monday of the month in which IACUC review is requested. If the first Monday falls on a University holiday, the deadline is the first working day that follows.
Expedited Review – Under exceptional circumstances, a Principal Investigator may request an expedited application review (3.0 IACUC Review Process Section 1.5.5 of the IACUC Policies and Procedures). The granting of an expedited review is rare and is subject to a processing fee. Missing a regular monthly submittal deadline is not in itself sufficient reason to request an expedited review
Section A—Administrative Information
1. Principal Investigator
The faculty or staff member responsible for the research project. Must have the authority to make decisions in the event of emergency or catastrophe situations. The principal investigator does not have to be the same as the grant funding the research.
2. Project Title Provide a descriptive title for the project. The protocol title need not match the title of any grants to which it will apply.
3. Submission Type Check one box to indicate the type of Application:
New Application – A first-time submission for a protocol.
Renewal– A submission to renew a previously approved protocol that is about to expire. A complete Application is required regardless of whether any procedures have changed. Provide the previous protocol number and give a brief explanation of what has been completed in the previous period and what will continue into the renewed protocol. Indicate any changes in the experimental design that are suggested by your results so far. This section is intended to help the IACUC understand the relationship of your expiring protocol to the renewal, and is not a request for a scientific "progress report."
Modification to open protocol–Make requests for modifications to a protocol by checking the Modification to open protocol and provide a brief explanation summary of the changes requested and the reasons(s). Use the track/change feature in Word to incorporate all necessary changes throughout the document and then send an electronic version to the IACUC office. Submit only section G for personnel changes. Once the changes are approved a signed version will be requested. This provides both the IACUC and your laboratory personnel with an up-to-date version of your protocol.
4. Special Categories Check the appropriate box or multiple boxes if any of the following apply:
Breeding only – Protocols intended solely to generate animals for research or teaching use are classified as breeding protocols. The actual experimental procedures or teaching use of the animals must be covered by other approved protocols – if a protocol includes a research or teaching component, this box should not be checked.
Teaching only – Protocols covering animal use for teaching purposes only are classified as teaching protocols. If the protocol includes a research component, this box should not be checked.
Pilot study – Protocols intended to allow the investigators to perfect new techniques or to obtain preliminary data to be used in the design of more complete experiments. A pilot study typically involves few procedures and a small number of animals. Some uncertainty in the experimental and statistical designs is allowable. The approval period is generally short (one year); requests for a longer approval period or a renewal must be justified.
5. Protocol Duration
Protocol duration begins on the date of approval by the IACUC and continues for the number of years requested in this section. An approved protocol is valid for a maximum of three years and is reviewed at least annually for the period requested. Protocols may be closed early by contacting the IACUC office. Projects running longer than three years will require submission of a renewal Application before the end of the initial three-year approval period.
6. Grant Support If known, list the funding agency supporting the research, the title of the grant, the submission date and the effective date. If the protocol will be supported by more than one grant, provide information about each grant. The grant title(s) need not match the protocol title. Please submit only one Application for any set of experiments, even if you will seek funding from several sources. 7. Record Keeping Identify all locations where records will be kept during the protocol period, and how they can be made available for inspection by IACUC, USDA, AAALAC or PHS staff members if necessary.
8. Principal Investigator Certification The Principal Investigator must sign the certification before the IACUC will review the Application. The signature of the Principal Investigator on the Application indicates that the Principal Investigator has read and understands the information and assurances contained in the Application and that it is an accurate description of the proposed use of animals.
This section should include a short description of the research aims, and explain the scientific merit of the proposal. The use of animals should contribute to the enhancement of human or animal health, the advancement of knowledge or the good of society. Language and terminology must be understandable by a member of the general public, for example, the community representative on the IACUC. Technical terms should be avoided when possible, and explained when they are indispensable. Language copied from a grant application is usually inappropriate.
2. Justification for Animal Use and Species Choice Two issues must be addressed in this section:
i. Explain why it is necessary to use live vertebrate animals in this project rather than an alternative study system. The Animal Welfare Act requirement to replace animals in part or full with non-animal systems or with less sentient animal species or stages when possible must be addressed. Non-animal systems include cell cultures, microorganisms, plants, in vitro systems, and computer and mathematical models. Less sentient alternatives to vertebrate animals include invertebrates and eggs of birds and reptiles. When a research question can be meaningfully pursued using reasonably available alternatives, these alternatives should be chosen.
ii. Justify the selection of the particular species and strain (if appropriate) requested. The most appropriate species/strain for the project based upon anatomical, physiological, or other characteristics in consideration of the scientific objectives and the need to obtain valid results should be chosen. This information is required to assure that the animal model is appropriate. The species selected should be the least sentient one possible.
3. Outline of Experimental Design
This overview of the experiments should provide the context for the animal numbers and the procedures described later. The use of outline or tabular format is encouraged. Include information on experimental and control groups, a list of procedures to be performed on each (using the procedure names in Section E 6, 7, or 8) variables to be measured, and the endpoint for each group. It is very important to list the time intervals between the beginnings of the animal’s role to the end of the experiment.For designs that involve multiple procedures occurring at different times, please provide a clear timeline. A flowchart or similar diagram can also be helpful for more complex experimental designs. The details of the procedures should be provided in your response to Section E 6, 7, or 8, not here.
Have previous experiments answered the questions or tested the hypotheses on which your research focuses? If the study duplicates previous experiments in whole or part, check other and explain why the duplication is scientifically necessary. If there is no duplication of previous research, this should be stated along with a brief indication of how this conclusion was reached (for example database search, journal review, scientific conferences). An example would be, “Based on a database search of studies in this area, the proposed work does not duplicate previous studies.” You should retain a copy of the search results on which you base your answer, but currently the search need not be detailed in the Application. Applications for teaching or breeding need not explain the need for duplication.
Section C—Animal Numbers
1. Summary Animal Number Table
State the number of animals of each species requested. Provide separate totals for animals in the following categories as applicable. When possible use a separate table to show the math you used to determine the number of animals you are requesting (e.g. groups x number of animals x procedures =?). Rat and mouse pups as animals are not counted until they are weaned. However, if pre-weaned animals are to be used as experimental animals, please indicate the total number to be used. Also, the number of adult breeders should be based on the number of experimental embryos or pre-weaned animals needed. For USDA species the total animals including pups are to be counted.
Estimate the number of animals anticipated as replacements for animals lost or unusable due to failure of procedure or other factors. In the event of unanticipated losses after a protocol has been approved, additional animals may be requested by filing a protocol modification.
For breeding, state the numbers of weaned and adult animals that will be required to 1) maintain the breeding colony and 2) supply experimental animals to any protocols that will receive animals from the breeding colony. Include all breeders and weaned offspring that will be acquired or produced whether or not they will be used in any experimental procedures (for example, both desired and inappropriate genotypes). Additional animals can be added to the breeding protocol in conjunction with new experimental protocols that receive animals from the breeding colony by a modification request stating the increase in breeders and offspring (weaned or pre-weaned).
If the Application is a Renewal, for any ongoing experiments break down the total number of animals needed into a) animals already used, b) animals currently in use, and c) additional animals requested for the work covered by the Renewal Application.
2. Justification of Animal Numbers
The Animal Welfare Act requires that you use the minimum number of animals necessary to obtain scientifically valid data through appropriate experimental and statistical design. Please justify the number of animals needed to accomplish your goal(s) and explain how you arrived at this number. Explain how the number of animals per group (or per time point) was selected. The justification should include a description of the statistical approach and an associated power test (with expected group differences and standard deviations) used to determine the minimum number required to obtain adequate statistical power. Statistical consultation may be obtained through the IACUC. You will find free power analysis programs are available on the IACUC webpage athttp://www.creighton.edu/researchcompliance/iacuc/sop/index.php.
If tissues are being collected for in vitro studies, justify the animal number based on the amount of material needed and the amount of material obtained from each animal (e.g. number of animals per antigen, or number of cells needed and number of cells obtained per animal) in order to obtain statistically valid results from the in-vitro study.
Breeding-only protocols do not need to justify animals to be transferred to experimental protocols (as this justification is provided in the experimental protocol.) Animals for other protocols are only approved after the other protocol is approved. However, the breeding protocol must justify the number of breeders based on the estimated yield of offspring of the desired genotype.
If the proposed work is a pilot study, indicate how the number of animals was estimated.
3. Acquisition and Primary Housing All Investigators are encouraged to consult with the ARF Manager to discuss animal housing before submitting an Application. All animal orders, regardless of animal origin and destination, must be made through the ARF. Prior arrangement with the ARF Manager is required to obtain animals from other than approved vendors. These animals must have a certification of health that is acceptable to the Attending Veterinarian and will normally be quarantined on arrival for a period determined by the Manager (see section 3.1.2 of the ARF Standard Operating Procedures).
Complete the table in the Application for each species or strain of animal to be used under the protocol. Do not include rat or mouse pups (pre-weanlings) in this table.
Species/Strain – Identify the species and strain (if known) as specifically as possible. For projects involving generation of specific genotypes (heterozygotes, recessive homozygotes, etc.), identification of the primary strain or mutation is sufficient.
Source – Supply the name of an institution if the animals are being obtained from another facility. For animals obtained from a commercial vendor use CV. For animals obtained from an in house breeding program, use the number of the appropriate breeding protocol “IACUC 0000.” For animals to be bred as part of the submitted protocol, use “bred.” Animals captured in the field should be labeled “wild-caught.” (Note: all wild-caught animals must be approved by the ARF Manager.)
Total Number from this source – Indicate the anticipated number of animals (other than mouse and rat pups) needed for each year of the protocol. The total number for each species or strain should match that indicated in question C1.
Approximate Daily Inventory – To facilitate planning by the ARF, indicate the expected number of animals needing to be housed at one time.
Primary Housing location – “ARF” may be used, unless animals are to be housed in an auxiliary animal room or laboratory. Housing in laboratory space is by special approval of the IACUC only.
Section D—Animal Husbandry 1. Animal Care Check the appropriate box to indicate who will provide care for the animals. If non-standard care is to be provided by ARF personnel, indicate the type of care they are to provide such as special diets or special care for animals with disease or functional deficits, prior arrangements must be made. Any special care needs and all care by non-ARF personnel must be described. Special safety precautions needed in handling of animals, cages or waste because of hazardous agents (see question F1) should also be indicated.
2. Animal Health
State how animal health, including pain and distress, will be assessed and specify the plan of action in case of animal illness or injury. Indicate any specific situations in which you wish to be notified prior to initiating treatment by the ARF staff. State the criteria (for example, tumor size, percentage body weight gain or loss, inability to eat or drink, behavioral abnormalities, clinical symptoms, signs of toxicity) for terminating procedures and/or for euthanasia if your experiments involve tumors, biologics, infectious agents, radiation or toxic chemicals. Provide scientific justification if animals will not be euthanized when moribund. It is not necessary to repeat methods for monitoring and treatment following experimental procedures, which should be discussed in sections E 6, 7, or 8.
3. Deficits in Transgenic and Mutant Animals
Genotype: Describe any post-natal functional deficit that is known or is likely in transgenic animals.
Nature of deficit/plan of care: Describe any special care or monitoring that will be required to detect and alleviate pain or discomfort associated with the deficit. If there is no information available as to the phenotype of a novel transgenic strain, indicate how you will assess the health of the animals and the plan of action should deficits arise.
Category of pain and distress (see section E1): Indicate the category of pain and distress if the category is expected to be D or E.
Disposition of Surviving Animals
Indicate the disposition of any animals for which the endpoints have not been specified as part of the experimental procedures (for example, if breeding produces excess offspring or unneeded genotypes). If animals are to be transferred to another protocol, specify the protocol.