Preparing a Protocol for Submission
Information about creating and submitting a protocol for the use of vertebrate animals in research and obtaining IACUC approval is provided below:
Protocol Preparation Checklist
IACUC Protocol Preparation Checklist
Please review this checklist to ensure that your IACUC application materials are complete before you submit:
- The individual listed as Principal Investigator (Lead Researcher) is eligible to serve in that capacity. To serve as Principal Investigator (Lead Reseracher), a UCI faculty appointment of greater than or equal to 50% is required; otherwise, an eligible individual must be chosen and listed as a Faculty Sponsor. Contact your department regarding who may be your Faculty Sponsor.
- All study team members, the Principal Investigator, Faculty Sponsor, and those listed in the protocol must complete all Training Requirements (CITI, LAOHP).
- Individuals that will have direct, hands-on contact with animals, along with any individuals who may be named as co-authors on publications resulting from the research, should be listed in the protocol.
- All other required regulatory committee approvals have been obtained or are in process, such as:
- Institutional Biosafety Committee (IBC) – recombinant DNA, materials of human or non-human primate origin, toxins, select agents, etc.
- Human Stem Cell Research Oversight (hSCRO) – use of human stem cells (embryonic, induced pluripotent, etc)
- Radiation Safety Committee (RSC) – radioactive materials
- Institutional Review Board (IRB) – use of human subjects
- Your Animal Use Protocol (AUP) has been completed and submitted in the Research Management System (RMS)
Lead Researcher Eligibility
Lead Researcher Eligibility
Lead Researcher and Principal Investigator Differences
Principal Investigator eligibility for a Contract or Grant is separate from Lead Researcher eligibility for research with human subjects. In making this distinction, UCI recognizes both the added responsibility associated with research involving humans and the research contributions of all members of the University community. Therefore, there are separate, but related, criteria to determine eligibility for these activities.
If an individual meets the criteria for Principal Investigator by virtue of their University appointment status, s/he may also serve as Lead Researcher on a research protocol involving humans. If an individual does not meet the criteria for Principal Investigator s/he may serve as Lead Researcher if s/he obtains a Faculty Sponsor or the activity.
Who May Serve as Lead Researcher of a UCI Protocol Application?
Individuals serving as Lead Researcher (LR) on IRB, protocols must have a formal affiliation (i.e., a faculty or staff appointment; enrolled student) with UCI. The type of appointment an individual has with UCI determines whether they may serve as a Lead Researcher on their own or whether a Faculty Sponsor is required:
- Senate eligible faculty and academic administrators may serve as LRs of protocols.
- Non-senate faculty and certain researchers with paid appointments of 50% or more may serve as LRs of protocols.
- Any other UCI-affiliated individuals (e.g., students, volunteer faculty members and staff) may serve as LRs of protocols if they have a Faculty Sponsor whose appointment meets #1 or 2 above).
|Series and Titles||Principal Investigator Eligibility (1)||Lead Researcher Eligibility|
Assistant, Associate, or Professor
Assistant, Associate, or Professor
|Lecturer - Track & SOE
Lecturer w/ Security of Employment (SOE)
Assistant, Associate, or Professor of Clinical ______ (Dept. Name)
|Clinical (With Salary)
Assistant, Associate, or Clinical Professor
|Yes, if salaried at 50% or more||Yes, if salaried at 50% or more. If less than 50%, Faculty Sponsor is required. (2)|
Assistant, Associate, or Professor
Assistant, Associate, or Professor
|All other Series, Titles, Appointments and UCI Students||No (1)||No, except with a Faculty Sponsor. (2)|
(1) For exceptions, see Principal Investigator Eligibility on Sponsored Projects webpage
(2) The Faculty Sponsor must be eligible to be a Principal Investigator.
Consultation with ULAR Veterinarians is strongly encouraged and should take place early in the protocol development process, well before the application is submitted for IACUC review.
Veterinary assistance is particularly helpful in the following areas:
- Animal model and species selections
- Refinement of surgical and non-surgical techniques
- Therapeutic drug regimens
- Appropriate use of sedation, anesthesia and analgesia
- Pre-, peri- and post-operative care and monitoring
- Specialized husbandry needs
- Availability of vivarium space
ULAR Veterinarians can also provide:
- Guidance regarding study design and appropriate techniques
- Guidance for alternative procedural endpoints and humane euthanasia
- Training in appropriate handling and care of animals
- Assistance in transferring animals from other institutions
- Assistance in development of environmental enrichment to promote psychological well-being of higher species
See Veterinary Services for more information.
Application Writing Tips
IACUC Application Writing Tips & Guidance
Tips for Writing Your IACUC Protocol
- Provide a summary of the project in lay language in the Project Overview section (General Information Tab). The project overview needs to be written in a manner that is lay-friendly and can be easily understood by a NON-scientific audience with little to no scientific background - Imagine you were describing your research to a classroom of students.
- Consider the “Three Rs” and unnecessary duplication (Rationales & Alternatives Tab):
- Replacement: Are there any alternatives to the use of live animals? If yes, you must provide a scientific justification for not using them.
- Reduction: Is the number of animals requested in the protocol adequate to achieve scientifically-valid results? How did you determine that the number is adequate but not unnecessarily large?
- Refinement: If procedures are likely to cause pain or distress, what will be done to minimize it?
- If similar projects or experiments are found in the scientific literature, describe what it is that makes this project unique.
- See the Alternative Searches Guidelines for additional guidance.
- Be sure all study team members are listed in the personnel categories according to the RMS permissions in which they need (Personnel Tab)
- View Protocol Only (Other Research Personnel) = default permissions
- Animal Ordering (Research Personnel)
- Protocol Editing (Co-Investigator/Senior Researcher, includes animal ordering)
- Provide justification for why a particular species was chosen as the animal model and describe any strain or breed characteristics. If multiple strains or breeds of a species is needed, explain why (Species Tab).
- State your hypothesis and design experiments to test it (Experimental Design Tab). Be sure to provide:
- Scientific rationale for all experiments
- Timeline of procedures that will be performed on the animals
- Post-procedural monitoring performed by the study team (not ULAR)
- Experimental Endpoints (point at which the live-animal portion of the experiment is complete and animal is euthanized)
- Humane Endpoints (criteria that are used to euthanize an animal earlier than the expected experimental endpoint)
- List the animal numbers, specifying the different groups and # per group and provide justification for how the number of animals was determined (Animal Numbers Tab)
See additional guidance regarding animal numbers
- Provide details of all procedures (both non-surgical and surgical) that will be performed on the animals (Procedures Tab).
- Discuss in detail any clinical signs and symptoms, possible side effects and adverse effects associated with either the disease model being studied or the procedures to be performed (Animal Monitoring & Other Info Tab). Describe how animals will be monitored and treated for pain, distress and discomfort, and list the criteria that will be used for determining that animals must be removed from the study and euthanized.
- List all agents and drugs that will be administered to live animals (Drugs & Other Agents Tab).
NOTE: All items listed in this tab should also be referenced in other relevant sections of the protocol (Experimental Design, surgical/non-surgical procedures, pain management, etc.).
- Describe how animals will be euthanized at the end of the study (Experimental Design Tab, Procedures Tab). Death must be confirmed in all animals prior to carcass disposal; in addition, UCI IACUC Policy requires that all rodents euthanized via CO2 overdose undergo a secondary physical method of euthanasia (e.g., decapitation, cervical dislocation or exsanguination) prior to disposal to prevent unintended recovery. See the IACUC Policy on Euthanasia for more information.
- Names (researchers, collaborators, etc.) should only be listed in protocol sections that specifically request for this information (Personnel Tab, Training Requirements Tab). In the event an animal use protocol is subject to a public records request, names are redacted for protection and names appearing unnecessarily in other sections may potentially be missed. Note: author names of publications can be included, since those are already public.
Additional Details about Specific Procedures
As you enter information into the protocol tabs in RMS, you may be prompted for additional details, depending on your specific experiments and proposed procedures. The following list includes items that the IACUC will require specific details about:
- Project Continuation: Essentially a progress report; Required ONLY if you are writing a three-year de-novo review of an ongoing, project previously approved by the IACUC
- Breeding: Required if a breeding colony of animals will be maintained to supply offspring for research purposes (usually genetically-modified rodents).
- Prolonged Restraint: Required if animals will be restrained for more than very short periods of time while awake. See the IACUC Policy on Animal Restraint.
- Use of Paralytic Agents: Required for the use of neuromuscular blocking agents in anesthetized animals.
- Multiple Survival Surgery: Required when individual animals will recover from more than one surgical procedure. Review the IACUC Surgery Policy and Guidelines.
- Antibody Production: Required for the production of monoclonal or polyclonal antibodies in live animals. Further information on the applicable policies is available.
- Non-UCI Locations: Required if research procedures with live animals will be conducted at facilities other than the UCI Campus or UCI Medical Center.
- Researcher-Maintained Animals: Required if laboratory staff will provide care and husbandry of the animals either within ULAR-operated housing facilities or outside of vivarium housing.
- Housing Exception: Required if animals will be kept outside of ULAR-maintained vivarium housing space for more than 12 hours.
- Variations in Husbandry/Housing: Required for all variations to standard ULAR-provided husbandry and care. Examples include use of wire-bottom caging, food or water restriction or additions, variations in standard light cycles, temperature or humidity, etc.
- If your research involves Hazardous Agents, you may be required to submit various documents pertaining to their safe use, such as Animal Room Hazard Communication Forms (door signs), Material Safety Data Sheets (MSDS) and Standard Operating Procedures (SOPs) for the handling of hazardous substances.Contact EH&S for assistance - See the Vivarium Safety website for more information.
- The IACUC performs a comprehensive pre-review process – if changes or additional information are necessary, you will have an opportunity to provide it prior to the convened meeting at which the protocol will be discussed.
Consideration of Alternatives to Animal Use
Alternatives Search Guidelines
Federal Animal Welfare Regulations1 require research institutions to ensure that investigators have appropriately considered alternatives to procedures that can cause more than slight or momentary pain or distress in animals, consistent with sound research design. Federal regulations1 require:
- 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 narrative description of the methods and sources, e.g., the Animal Welfare Information Center, used to determine that alternatives were not available.
- The principal investigator has provided written assurance that the activities do not unnecessarily duplicate previous experiments.
The U.S. Department of Agriculture has established an information service at the National Agricultural Library2 to assist researchers in documenting the consideration of alternatives to animal use, minimization of pain and distress and avoidance of unnecessary duplication of experiments.
"The Three Rs"
The consideration of alternatives is framed within the context of the "3 R's" originally discussed in The Principles of Humane Experimental Technique (Russell and Burch,1959):
Replacement: Utilizing non-animal models. These may include living systems utilizing organ, tissue or cell culture techniques; non-living systems such as chemical techniques and mechanical models for training; and computer simulations.
Reduction: Determining the number of animals used in a study by experimental design and/or statistical analysis to minimize the number of animals required to obtain meaningful data.
Refinement: Reducing or eliminating pain or distress in animals by use of anesthetics and analgesics, modification of procedures or technical enhancements that increase the quality or quantity of data gathered.
Guidance for Consideration of Alternatives
- Federal regulators (e.g., USDA and PHS) consider the performance of a computerized search of scientific literature databases to be the most effective and efficient method for demonstrating compliance with the requirement to consider alternatives and avoid unintended duplication.
- A thorough search of the scientific literature usually requires more than one database since no single database reviews all literature in all research fields.
- Alternative searches are required in the planning phase of the animal use protocol and are given equal consideration along with the overall experimental design and the appropriateness of the animal model to be used. Additional alternative searches should be performed when a protocol is changed significantly by modification and are required when the modification involves use of a new, more-invasive technique or procedure.
- All animal use protocols must provide written documentation of the researcher’s good faith effort to determine the availability of alternatives and consider their applicability to the specific research project.
Required Documentation for the Consideration of Alternatives
Principal Investigators or study designers must provide the following as part of the written protocol narrative:
- The name(s) of the database(s) searched.
- The date when the searches were performed (e.g., "9/25/05"). NOTE: This date may not be more than three months prior to submission of the application to the IACUC for review.
- The date range over which the search was conducted (e.g., "1950 - present").
- The search terms or keywords used to search the databases. Appropriate search terms should reflect the animal model, scientific design, proposed experimental procedures, etc.
- A detailed narrative description of the results and findings of the searches, including an explanation for why any alternatives found but not used are not appropriate for the study.
In some circumstances, consultation with subject experts or participation in scientific conferences or colloquia may be used to support the consideration of alternatives. In such cases, detailed information about the consultation or conference should be provided (e.g., date and location of conference, name and qualification of consultant, etc.)
The UCI Libraries staff can assist you in the successful performance and documentation of literature database searches alternatives:
Additional guidance is available from the National Agricultural Library, Animal Welfare Information Center (AWIC):
Determination and Justification of Animal Numbers
Determination and Justification of Animal Numbers
A key principle governing the ethical use of animals in research, testing and teaching is that no animal life is wasted; the number of animals used in each project must be the minimum necessary to obtain valid and meaningful results1.
By Federal regulation, the IACUC must review the number of animals requested in each protocol and agree that the number is appropriately justified in terms of the stated goals of the project.2,3
Experimental Design Outline
Justification of animal numbers begins with a clearly-stated, hypothesis-driven outline of the experimental design. All experiments involving live animals or animal tissue must be outlined in the protocol, and the number of animals required must be directly relevant to the experimental design in order to achieve the goals of the project. All animals must be counted and justified, including experimental animals, donor animals and live-born offspring of pregnant animals and animals that are produced in breeding colonies but cannot be used in research.
NOTE: Animals used to establish and maintain a breeding colony (breeding pairs, offspring that cannot be utilized because of genotype, sex, etc.) should be justified separately in Appendix B, along with a justification for maintaining the colony. The total number of animals justified in Appendix B (including all animals that will be produced in the colony but not used for any purpose) should then be LISTED in Application Section H. Breeding animals do not need to be included in the Experimental Design section.
In order to assess the number of animals requested, the Lead Researcher must provide the IACUC with an overall outline of the proposed experiments in the protocol application. This outline must include (at minimum):
- The purpose of each experiment or set of related experiments,
- The number of experimental groups/subgroups;
- The number of animals by species/strain per group/subgroup,
- The total number of control and experimental animals should be drawn from the experimental design and summarized in the overall outline. For many studies, this information can be effectively presented using a table or grid design.
- If the project involves euthanizing animals for tissue harvest, the relationship between the amount of tissue/cells needed for the experiments must be directly correlated to the number of animals required to produce that tissue (e.g., each animal produces X amount of tissue, which is adequate to perform Y experiments.)
- Anticipated animal losses due to morbidity, mortality or other expected difficulties with the experimental procedures must be carefully described in order to justify the need for additional animals.
Justification of Animal Numbers
Research and testing studies should be designed to provide a statistically significant result with a minimum number of animals, and the method by which the number of animals was determined must be clearly stated. Statistical techniques and/or power analysis are appropriate in most cases to maximize the analysis of the data generated from each animal. However, the IACUC acknowledges that the basis for an appropriate justification of animal numbers depends largely on the nature of the study itself. Prior experience and expertise with the model in question may be taken into account as well, but must be carefully documented in the protocol.
Consultation with a statistician or use of statistical software during the design phase of the experiment may be useful. The UCI Center for Statistical Consulting can provide assistance in estimating animal numbers in research proposals.
Websites that may be helpful in performing a power analysis include:
Five basic types of studies are listed below, along with guidelines for justification of animal numbers appropriate for each type of study. These guidelines are intended to provide direction – any given study may not fall neatly into one of these five categories:
Exploratory Study Requiring No Statistical Analysis (use of live animals to demonstrate success or failure of a desired goal, such as the production of transgenic mice): Animal numbers are justified based on the probability of success of the experimental procedure; a detailed explanation of how that probability was determined must be included in the narrative.
Pilot Studies: Animal numbers are determined by the investigator’s experience and personal judgment, and are typically small. Data collected in pilot studies are generally used to determine statistically relevant sample size calculations for future experiments.
Studies Requiring Inferential Statistical Analysis: If possible, animal numbers are determined by statistical power analysis; the justification statement must include the values of alpha, beta, sigma, and effect size used in the power analysis to determine sample size. Alternatively, minimum numbers of animals may be determined based on pertinent literature for comparable studies in which the desired effect sizes were shown to be statistically significant.
- Animal numbers cannot be justified on the basis of how many experiments the lab personnel can perform in a week, month, etc.
- The cost of the animal should not be considered as the primary justification for the use of a particular species or model.
- Public Health Service. (1996) U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and training. PHS Policy on Humane Care and Use of Laboratory Animals. Washington, D.C.
- National Research Council. (2011) Guide for the Care and Use of Laboratory Animals, Eighth Edition. National Academy Press, Washington, D.C.
- CFR (Code of Federal Regulations) (1985) Title 9 (Animals and Animal Products), Subchapter A (Animal Welfare). Washington, D.C.: Office of the Federal Register.
- ILAR Journal Volume 55, Issue 3, 2014 - Experimental Design and Statistics https://ilarjournal.oxfordjournals.org
- Erb, H.N. (1996) A non-statistical approach for calculating the optimum number of animals needed in research. Lab Animal, 45-49.
- Mann, M.D., Crouse, D.A., Prentice, E.D. (1991) Appropriate animal numbers in biomedical research in light of animal welfare considerations. Laboratory Animal Science 41:6-14.
Record-Keeping and Reporting of Animal Numbers
All researchers understand the importance of accurate and copious records - they are the backbone of your research endeavors and the basis for scientific publications, grant proposals, etc. In addition to laboratory notebooks and other data-based research records, researchers that use live animals should also maintain records of:
- Training for all personnel who work with live animals.
- Surgical and post-operative monitoring records.
- Post-procedural monitoring records (e.g., water/food restriction logs)
- Number of animals used for research purposes
- These are animals that undergo experimental procedures and/or manipulations (e.g., behavior tests, drug administration, euthanasia for tissue collection, etc.)
- Records should include how the animals were used and the relevant USDA pain category (C, D, or E)
- Includes exotic species not purchased through ULAR, wild-caught animals, etc.
- Total number of animals produced in breeding colonies but not used in experiments. These animals are all USDA pain category B and the total number includes:
- Animals held as replacement breeders
- Animals that are euthanized immediately prior to weaning (e.g. culled littermates)
- NOTE: Animals produced in the breeding colonies that are used in experiments are to be counted as "used for research purposes" in the relevant USDA pain category.
Guidance for Reporting Animal Numbers:
The following animal numbers are recorded in RMS, and deducted from a protocol's approved allocation.
- Animals purchased through ULAR
- Animals transferred by ULAR (e.g. from other institutions, or between protocols at UCI)
The number of animals produced in campus breeding colonies or obtained by other methods (e.g. wild-caught, obtained from non-traditional sources) are not recorded in RMS and must be recorded by the researcher and reported to the IACUC at least once per year.
- While the IACUC does not require any specific method or software for maintaining breeding colony records, accurate records are absolutely required for both scientific and animal-welfare reasons. See IACUC Policy on Rodent Breeding Colonies.
- To avoid double-counting animals, you should omit the number of breeder animals currently in the breeding colony if they were already reported during a previous protocol reporting period. See guidance for how to look up previously-submitted Annual Reviews in RMS.
Reporting Animal Numbers via the Annual Review:
At the end of each protocol year, Principal Investigators or their designees must submit a progress report (Annual Review) for the reporting period. Reporting periods begin with the protocol approval date:
- End of Year 1 = IACUC approval date + 365 days = Annual Report due
- End of Year 2 = IACUC approval date + 730 days = Annual Report due
- End of 3-year protocol period = IACUC approval date + 1095 days = Submit rewritten protocol for full committee review.
Each Annual Review submission should include the number of animals used in research during the last protocol year, including:
- Number of animals purchased or transferred through ULAR and used for either research purposes or as breeding animals. NOTE: Report all animals in the appropriate USDA Pain Category B, C, D or E -- the highest pain category according to how they were usedaccording to how they were used
- Number of offspring produced in the colony and used for research purposes (these animals should be reported in the appropriate USDA Pain Category C, D, or E)
- Number of offspring produced in the colony and maintained as replacement breeders (Category B)
- Number of offspring produced in the colony but not used for any purpose, such as culled littermates (also Category B)
As part of the administrative processing of Annual Review information, IACUC staff will compare the number of animals reported by the lab to the total number of animals counted against the approved allocation in RMS and update the RMS totals as necessary.
IACUC Pre-review Process
IACUC Pre-review Process
All submissions (protocols, modification requests) that undergo full committee review go through a detailed pre-review by IACUC administrative staff, campus veterinarians and voting members of the IACUC.
- Submission deadlines occur 16 business days prior to the monthly convened meeting (the second Thursday of each month) to allow time for detailed pre-review.
- Administrative staff, veterinarians and committee members review and comment on all submissions during the first week of the pre-review period.
- Pre-review comments are compiled and forwarded to Lead Researchers approximately 7 business days prior to the scheduled meeting.
- Research personnel have until the end of the day Monday of meeting week to submit revised materials for review at the convened meeting.
- Protocols and modifications with serious issues noted in pre-review may be removed from the agenda and tabled to a future meeting if revised documents are not received prior to the meeting.
All submissions to the IACUC undergo pre-review by IACUC administrative staff to determine the appropriate level of review. NOTE: Level of review is determined administratively with input from campus veterinarians and the IACUC Chair wherever necessary. At some points in the review cycle, a modification request that might ordinarily qualify for Designated Member Review may be placed on the agenda for full committee review; in such cases, IACUC administrative staff has determined that review at a convened meeting will result in shorter approval turn-around.
Administrative and Veterinary staff is also available to review and comment on protocols and modification requests prior to formal submission, and can advise researchers on questions regarding the level of required review.
See the IACUC Policy on Modifications for more information.
Top 9 Reasons that IACUC Submissions are NOT Approved
Top 9 Reasons that IACUC Submissions are Not Approved
Not enough information
The protocol does not clearly express either the rationale behind the experiments or the justification for the use of live animals. Be sure to review all sections of the protocol narrative application and provide ALL requested information.
Too much information (in the wrong place)
The Experimental Design section must provide an overview of the project as a whole, including the rationale for the experiments, a summary of the experimental groups and what procedures will be performed on them, and the timeline for those procedures (when they are performed in relation to other procedures and to the endpoint of the experiment). Details about the procedures themselves do not belong in the Experimental Design section, but it should be clear exactly what will happen to each of the animals and why.
The Project Overview is too technical
The Project Overview is meant to be a lay summary of the project as a whole. It should be clearly understandable to people with little or no background or experience in your area of expertise. Leave out the jargon and technical terms, and provide an explanation of what you plan to do that is suitable for a newspaper article or non-scientific cocktail party chatter.
The consideration of alternatives to the use of live animals has not been documented
The IACUC understands that review of the scientific literature is an ongoing part of the research process; however, it must be documented in the protocol in order to meet the requirements of USDA Animal Care Policy #12. The IACUC Protocol Application form has been specifically designed to guide researchers in appropriately documenting the consideration of alternatives.
Animal numbers are not adequately justified
The number of animals requested for the experiments must be adequate to achieve the research goals of the study – researchers must provide an explanation for how the number of animals was determined. Wherever possible, statistical power analysis should be employed.
Inconsistencies in the narrative
All sections of the protocol narrative must be consistent – procedures and agents mentioned in the Experimental Design must be described in detail in the appropriate sections of the protocol narrative; animal numbers and pain categories must be consistent throughout the protocol, etc.
Animal Welfare Issues
If an equivalent procedure is reasonably expected to cause pain, distress or discomfort in a human being, you MUST assume that it would also cause pain, distress or discomfort in an animal and address these issues accordingly in the protocol narrative. Monitoring and assessment parameters must be described in detail, and relief from potential pain and distress must be provided unless there is a strong scientific justification for withholding it.
Modification Mishap #1 - Proposed changes not incorporated into the protocol
IACUC policy requires that the protocol be updated with each approved modification, which means that the proposed changes need to be incorporated into the relevant protocol tabs and section.
Modification Mishap #2 - Mod doesn’t fit in with the existing approved narrative
Proposed changes that significantly alter the originally approved protocol may be more appropriately submitted as a new stand-alone protocol.