IACUC Protocol Application IACUC #:

Saint Mary’s College of CaliforniaInstitutional Animal Care and Use Committee

e-App #:

Mod #:

For Office Use Only

Provide your responses (as applicable) in the blank white sections provided. If a section does not apply to the study, state “N/A”. When marking checkboxes, you must place an X to indicate your selection(s). Any additional requirements that must be completed are indicated by yellow sections.

Select one:Protocol #:

3-Year Renewal Application:

Modification of an Approved Protocol:

New Application

Application Section

A. Title of Study

B. Principal Investigator

Name: Dept:

Phone #: After-hours Phone #:

Email:

C. Alternative Emergency Contact Person Emergency contact must be someone other than the Lead Researcher with the authority to make decisions about animal care.

Name: Home Dept:

Phone #: After-hours Phone #:

Email:

D. Administrative Contact Person

Person(s) listed here will automatically be CC’ed on all email correspondences to the PI

Name Email

Name Email

CITI Training Completed?

Refresher training required every 3 years

Yes No

E. Study Funding and Scientific Merit Review

Place an X in the checkboxes that apply to indicate how the study costs will be supported. This includes any pending funding sources. For pending funding sources, list “Pending” in the Award/Proposal # column.

Grant/Sub-award (list specific details below*)

Contract/Sub-contract (list specific details below*)Department or campus funds(e.g., department support, unrestricted funds, start-up funds, personal funds, campus program awards, etc.)Non-cash support from manufacturer/sponsor (e.g., free drug, device, research materials)

1. *List all extramural proposals or awards that will support this study:

Agency/Sponsor Award or Proposal # PI of Award

2. Have the experiments described in this protocol application undergone peer review by the funding agency/sponsor listed above?

Yes No

F. Animal Numbers

Based on the experimental design in this study, provide the following information about the animals in this project:

Genus-Species or Common Name

List the genus-species or common name of all vertebrate animals involved in the experimental design.

Condition CodePreg Animals purchased while pregnantDWL Dams with a litter

NA Not Applicable: This includes all other animals, regardless of age, under this code.

USDA Pain Categories

B Animals held only for breeding and not used for any research purpose

C Momentary or no pain, distress or discomfort (e.g., needle stick)

D Procedures reasonably considered painful or likely to cause discomfort or distress, but alleviated with analgesics, anesthesia or timely euthanasia.

*E Unrelieved pain, distress or discomfort (requires scientific justification)

Total # Required Indicate the total number of animals involved in the project for the duration of the project or 3 years, whichever is less

Each animal should be counted only once in this table. The most invasive or potentially painful procedure determines the USDA pain category.

Do not list Individual strains or subspecies here. Please include complete information about different strains, genetically modified animals, etc., in Section 3.

Species Condition Code(N/A, Preg, or DWL)

USDA Pain Category(B, C, D, or E*)

Total # Required

*Animals in Category E: Unrelieved pain, distress or discomfort requires strong scientific justification with particular attention to the significance, necessity, and potential benefits of the research.

Investigators must perform euthanasia on all moribund experimental animals unless there is scientific justification that euthanasia would invalidate experimental data collection.

If euthanizing a moribund animal would seriously harm the study, the scientific justification for using death as an endpoint must be provided below.

For additional information about SMC’s euthanasia policy and alternative endpoints, visit the IACUC page

G. Procedures to be Performed in Live Animals

List the species in which procedures will be performed.

Procedures

Species

Place an X in the checkboxes below to indicate the procedures that will be performed in those species. Complete any additional appendices as required.

1. Administration of experimental agents

2. Antibody production/collection (monoclonal & polyclonal)

3. Breeding

4. Blood/tissue collection in live animals

5. Behavioral tests

6. Euthanasia followed by tissue harvest

7. Invasive field study (Study of animals in their natural habitat involving an invasive procedure or one that harms or materially alters the animals’ normal behavior.)

8. Any variations in standard housing or husbandry (e.g., food/water restriction, special diet/water, alteration of cage environment, withholding of standard environmental enrichment, etc)

9. Major survival surgery (Surgical procedures which either penetrate a body cavity or result in permanent impairment of normal functions.)

10. Minor survival surgery

11. Multiple major survival surgeries

12. Use of neuromuscular blocking agents (Agents causing paralysis of skeletal muscles i.e. pancuronium, suxamethonium.)

13. Use of prolonged restraint (Greater than momentary manual or mechanical limitation of animal’s normal movement.)

14. Terminal surgery (Animal is not recovered from anesthesia.)15. Cardiac perfusion (Terminal procedure involving replacement of blood with fixative while animal’s heart is still beating.)

H. Hazardous Agents

Place an X in the checkboxes that apply to indicate which categories of hazards will be used in the study.

1. Anesthetic Gases

Anesthetic Gases

a. b. List the specific agent(s):

c. d. Explain the method of administration (e.g., vaporizer, bell jar, etc):

e. f. Describe the method used to capture waste gasses:(e.g., active scavenging, passive scavenging with F/Air canister, chemical fume hood, etc.)

2. Biological Hazardous agents (requiring approval for Biosafety)

Infectious agents (e.g., bacteria, viruses, fungi, etc.)

Human or primate materials (established cell lines, primary cells, tissue, fluids, blood)Recombinant DNA (lentiviral vectors, AAV, plasmids, etc.)

Select Agents/Toxins (as defined by the CDC Select Agent Rule)3. Chemical Hazardous agents . Note: The PI may need to submit a Standard Operating Procedure as part of the

Chemical Hygiene Plan while using this chemical with animals.Carcinogens, toxins, investigational new drugs and other potentially hazardous chemicals or agents (e.g., formaldehyde, tamoxifen, BrdU, etc.)

SOP Complete? Yes

Nanomaterials SOP Complete? Yes

I. Controlled Substances

Refer to the U.S. Drug Enforcement Agency’s list of Controlled Substances A Controlled Substance Use Authorization (CSUA) must be in place before any drugs can be ordered.

Place an X in the checkboxes that apply to indicate which controlled substances will be used in the study. Provide the corresponding CSUA #.

Ketamine

Telazol

Sodium Pentobarbital, including any prepared solutions that contain it (e.g., Euthasol®, Nembutal®)

Buprenorphine

Butorphanol

Other (specify):

CSUA #:

J. Animal Housing and Procedure Locations

Place an X in the checkboxes that apply and complete any additional appendices as required.

1. List all location(s) where animals will be housed and/or used:

Indicate the building, room number, and biosafety level below.

Activity Building Room Number(s) Biosafety Level

(required only for areas outside of vivarium)

(if applicable)(ABSL1, ABSL2, ABSL3)

Animal Housing

Non-surgical Procedures

Animal Surgery

Indicate husbandry and housing responsibilities:

Laboratory staff to provide routine animal husbandry and care in the animal facility (e.g., lab) Complete Appendix I

If laboratory staff are required by Hazardous Agents SOP (see Section J above) to provide animal care/husbandry following administration of hazardous agents, check here and indicate the length of the requirement by checking the appropriate box:

48 hours after administration

72 hours after administration

Other (describe):

Will you be using the animals and the protocols listed for teaching, research or both?

Departmental Approval IACUC #:

Saint Mary’s College of CAInstitutional Animal Care and Use Committee

For Office Use Only

The Department Chair’s signature is required if the study will be performed under the auspices of a Department

If the Department Chair is a member of the research team on this application, the approval must be obtained from the next highest level of administrative authority (e.g., School Dean).

Department Assurance Statement:

By signing below, I hereby confirm that I have read the IACUC Application and Protocol Narrative and I certify that: The research is appropriate in design.

(i.e., the research uses procedures consistent with sound research design, the study design can be reasonable expected to answer the proposed question, and the importance of the knowledge expected to result from the research is known)

The Principal Investigator is competent to perform or supervise the study.

There are adequate resources and funds available to support performance of this research, including costs associated with animal husbandry and care.

Protocol #:

PI:Title of Study:

Printed Name of Department Chair Signature of Department Chair/ Date

Investigator Assurance Statement IACUC #:

Saint Mary’s College of CaliforniaInstitutional Animal Care and Use Committee

For Office Use Only

As the Principal Investogator, I have ultimate responsibility of the performance of this study, the ethical and humane care and use of the animals, and strict adherence by all study team members to IACUC requirements, all federal, state and local laws, and regulations and policies of Saint Mary’s College of California governing the use of animals in research, teaching and testing.

I hereby assure the following:

1. The information provided in this application is accurate to the best of my knowledge.2. I have performed the required alternatives searches and to the best of my knowledge, the experiments and procedures

described in this protocol do not unnecessarily duplicate previous experiments, unnecessarily use animals, or unjustifiably expose animals to potentially painful, uncomfortable or distressful procedures.

3. All experiments and procedures involving animals will be performed under my supervision or that of another qualified professional listed on this protocol.

4. I will ensure all study team members are qualified and authorized to conduct procedures involving animals under this protocol. This includes ensuring that all study team members, including myself have: Been (or will be) trained in handling and care of the involved species and approved procedures in this protocol, including

the proper use of anesthetics, analgesics and experimental agents, and euthanasia. Read and understand the procedures outlined in the approved protocol. Completed the CITI Training on animal care and use and other required training courses (as applicable).

5. Personnel will be allowed adequate time to obtain necessary training for this project and will not begin any procedures with live animals until they have been successfully trained.

6. All animal acquisition will be housed in the Animal Facility in Brousseau Hall.7. Emergency veterinary care is permitted for animals showing evidence of pain or illness not addressed specifically in the

approved protocol, in addition to routine veterinary care as prescribed for individual species.8. All study team members are aware of the potential hazards associated with the use of live animals and animal tissues. In

addition, I understand I am responsible for notifying EH&S if an individual’s level of animal contact changes, due to changes in the protocol or job description.

9. I understand that, should I use the project described in this application as a basis for a proposal for funding (either intramural or extramural), it is my responsibility to assure that the description of animal use in the funding proposal(s) is identical in principle to that contained in this application. I will submit modifications and/or changes to the IACUC as necessary to assure these are identical.

10. I and all of the co-investigators and research personnel agree to comply with all applicable requirements for the use of animal in research, testing and teaching including, but not limited to, the following: Making no changes to the approved protocol without first having submitted those changes for review and approval by the UCI

Institutional Animal Care and Use Committee; and Promptly providing the IACUC with any information requested relative to the project; and promptly and completely complying

with an IACUC decision to suspend or withdraw its approval for the project; and Obtaining continuing review prior to the date approval for this study expires (I understand if I fail to apply for continuing review,

approval for the study will automatically expire, and study activity must cease until IACUC current approval is obtained); and filing a final report with UCI IACUC at the conclusion of this project.

Protocol #:

PI:Title of Study:

Printed Name of Principal Investigator Signature of Principal Investigator Date

IACUC Protocol Narrative

Saint Mary’s College of CaliforniaInstitutional Animal Care and Use Committee

Section 1: Project Overview

Provide a non-technical, lay language summary of the proposed project that can be understood by non-scientists, community members, and IACUC members with varied research backgrounds. Discuss the potential relevance (e.g., benefits) of research findings to human or animal health, advancement of knowledge, and/or for the good of society. Address the scientific merit of the research in this summary.

Avoid using scientific terminology, jargon or unexplained abbreviations; where use of such language is unavoidable, provide definition of the term where first used.

Do not cut and paste from a grant application, journal article or abstract. This summary should not exceed 250 words.

Section 2: Justification for the Use of Animals

Federal regulations require that investigators provide a narrative describing the rationale for using animals, the appropriateness of the species, and the methods and specific sources used to determine that alternatives (e.g., replacement, reduction, refinement) to the use of animals and to the procedures have been considered.

A. Rationale for the Use of Animals

1. Explain why animals are required for these studies, and why non-animal model replacements, such as cell culture or computer modeling, cannot fully replace animals:

2. Explain why the proposed species are the most appropriate:

B. Consideration of Alternatives to Live Animal Use, Painful Procedures and Unnecessary Duplication

The United States Department of Agriculture (USDA) requires that you specify at least two sources or databases used to determine that the model and methods described in this protocol do not:

Unnecessarily duplicate previous experiments, Unnecessarily use animals, or Unjustifiably expose animals to potentially painful, uncomfortable or distressful procedures

1. Database Searches

Place an X in the checkboxes that apply to indicate which databases were used:

National Agricultural Library Web of Science PsycINFO

(AGRICOLA) AltWeb TOXLINE

BIOSIS Previews CORDIS MEDLINE via PubMed

CRISP NORINA Web of Science

TBASE2. Date(s) the database search was performed:(Must be within the last 3 months)

3. Years covered by the search (e.g., 1985 to present):

4. Keywords used in the search:

The more "keywords" you use the more specific your search will be; however, being too specific may lead to no results being found.  In that case, reduce the number of terms used. Click here for search tips.

5. If a consultant was used for considering alternatives, provide the following information:

Expert Consultant’s Name:

Consultant’s Qualifications/Expertise:

Content of Consultation:

Date(s) of Consultation:

6. Database Search Narrative Description

Specifically discuss the results of what was found during the search. Describe the steps you have taken to replace the use of animals with in vitro procedures, reduce the number of animals used, and to refine the experimental design and procedural techniques.If similar experiments are found, describe the aspects of your research project that are novel and are not unnecessarily duplicative of other published work. If alternative methods or procedures representing refinements to your procedures were found, discuss why those alternatives cannot be used.

Discussion of Search Results:

Replacement:

Reduction:

Refinement:

Section 3: Experimental Design

Provide a concise description of the experimental design below, describing all experiments to be performed. For each experiment, provide the following information:

Rationale behind the experiment or the hypothesis being tested. Define the groups and number of animals per group needed for the experiment(s), including both experimental

and control animals and list the procedures to be performed on each group.  A table may be an effective way to present this information.

Explain how the number of animals for the experiment was determined to be adequate for the generation of statistically significant data (i.e.,, power analysis, previous publications).

Describe the sequence and timing of all live animal procedures to be performed, according to the groups defined above.  Provide a timeline, diagram or flowchart if appropriate.

Define the procedural endpoints for each group of animals defined in above (i.e. what determines when the live animal portion of the experiment is complete and animals are euthanized.)

DO NOT:- Cut and paste from a grant application, journal article or abstract.- Provide a detailed description of the procedures here. These details should be provided in Sections 5 and

6.- Provide detailed descriptions of analytical methods or procedures that do not involve the use of animals.

For best results, create your experimental design section in a separate word document (or other software) including all embedded tables, figures, etc. Once complete copy and paste it into the blank section below.

EXPERIMENTAL DESIGN BELOW

Section 4: Non-Surgical Procedures

Describe in detail all non-surgical procedures to be performed on live animals. Procedures described here must be referenced in Section 3. Animal monitoring and management (as applicable) must be addressed in Section 7. Each subsection below should be completed as it applies to the protocol. A veterinarian should be consulted in the planning of non-surgical procedures involving potential pain and/or

distress. If a particular procedure does not apply, state “N/A”.

Not Applicable - Non-surgical procedures will not be performed on live animals. (Skip this section)

A. Terminal Procedures 1. 2. Euthanasia followed by Tissue Harvest:

3. 4. Cardiac Perfusion:

B. Blood and Tissue Collection in Live Animals

C. Behavioral Studies

D. Administration Methods for Experimental Agents in Live Animals

Describe in detail the method by which each experimental agent will be administered in the animals.See Instructions. NOTE: Details about dosages and frequency of administration should be provided in Section 7.(To add more rows, place cursor in the last cell on lower right and press the key “Tab”)

Experimental AgentTotal Volumeof each Dose

(Including Vehicle)

Administration Procedure(See Instructions)

E. Induction of Anesthesia/Sedation for Non-surgical Procedures in Live Animals

1. Indicate how anesthesia/sedation is induced for non-surgical procedures:

2. Describe how the level of anesthesia/sedation is assessed to be adequate to begin the non-surgical procedure:

3. Describe how animals are monitored throughout the non-surgical procedure: e.g., respiration pattern, response to noxious stimulation (paw pinch, etc.), heart rate/pattern, EKG, blood pressure, temperature, etc.

4. Discuss any supplemental anesthesia dosing that may be required:

5. If animals will be placed on artificial ventilation, describe the range of respiration rates and tidal volumes.

6. For survival non-surgical procedures, describe how animals are monitored for recovery from anesthesia/sedation and when animals are returned to their home cages. Estimate how long it will take for the animal to recover from anesthesia/sedation (i.e., ambulatory and feeding).

F. Imaging Procedures in Live Animals:

G. Other Non-surgical Procedures in Live Animals:

H. Post-procedural Care for Non-Surgical Procedures in Live Animals (if applicable)

Section 5: Surgical Procedures

Describe in detail all surgical procedures to be performed. Any procedures described here must be referenced in Section 3. Animal monitoring and management (as applicable) must be addressed in Section 7. Each subsection below should be addressed as it applies to the surgical procedure. Address each of the applicable subparts for each type of surgical procedure that will be performed. If a particular procedure does not apply, state “N/A”

Not Applicable – Surgical procedures will not be performed on live animals. (Skip this section)

A. Pre-operative Care Describe any care given to the animals prior to the surgery (e.g., fasting, sedation, pre-operative physical exam or blood work, etc.)

B. Induction of Anesthesia for Surgical Procedures in Live Animals 1. 2. Indicate how anesthesia is induced:

2. 3. Describe how the level of anesthesia is assessed to be adequate to begin the procedure:

4. 5. Describe how animals are monitored throughout the procedure (e.g., respiration pattern, response to noxious stimulation such as a paw pinch, heart rate/pattern, EKG, blood pressure, body temperature, etc.). Discuss any supplemental anesthesia dosing that may be required.

6. 7. If animals will be placed on artificial ventilation, describe the range of respiration rates and tidal volumes:

8. 9. For survival procedures, describe how animals are monitored for recovery from anesthesia and when animals are returned to their home cages. Estimate how long it will take for the animal to recover from anesthesia (i.e.,, ambulatory and feeding).

C. Aseptic Techniques 1. 2. Preparation of the surgical space:

3. 4. Preparation of the surgeon (e.g., surgical scrub of hands, donning surgical attire, gloves, etc.):

5. 6. Preparation of the animal (e.g., clip fur, clean surgical site with antiseptics, use of sterile drapes, application of eye ointment, etc.):

7. 8. Sterilization of instruments (e.g., autoclave, glass bead sterilizer, chemical sterilant etc.):

D. Description of Surgical Procedures

E. Methods to Prevent Dehydration & Hypothermia

F. Surgical Endpoints

G. Post-Operative Care and Analgesic Usage 1. 2. Describe the post-operative care that will be provided to the animals following surgery:

3. 4. Explain the use of analgesics, including dosage and frequency:

Section 6: Adverse Effects & Animal Monitoring and Management

Describe the possible adverse effects that may arise during the course of this study. If a section is not applicable, state “N/A” NOTE: N/A only applies to experiments in which naive animals will undergo a terminal surgical

procedure or be euthanized for tissue harvest without any prior experimental manipulation.

A. Summary of Possible Adverse Effects or Phenotypic Abnormalities

Briefly summarize all possible adverse effects or phenotypic abnormalities that may present in the animals as a result of study procedures, agents, disease processes, genetic alterations, etc. (i.e., tumor formation, ascites, neurologic deficits, infection, diabetes, etc.)

B. Clinical Signs and Symptoms

Describe the clinical signs and symptoms that may appear in the animals as a result of the experimental procedures or agents, including any associated pain, distress or discomfort.

C. Monitoring Parameter

Describe the monitoring parameters that will be used to assess pain, distress and discomfort in animals (i.e., signs, symptoms and species-specific behaviors).

D. Management Plan Describe the management plan that will be used to assess and treat pain, distress and discomfort in the animals, including any special procedures that will be used (e.g., periodic weighing of animals) and any interventions that will be performed to relieve pain, distress and discomfort in the animals (e.g., analgesics, antibiotics, special housing/bedding, etc.).

E. Frequency of Animal Monitoring

F. Documentation of Animal Monitoring

Describe how the monitoring of animals (e.g., daily observations, treatments performed by research staff) will be documented.  Monitoring records should be kept in the animal room and must be readily available to ULAR veterinary staff and IACUC members at all times.

G. Euthanasia Criteria

Describe the criteria that will be used to determine that an animal must be removed from the study ahead of schedule and euthanized. (Click here for euthanasia policy and guidelines, including recommended criteria.)

Section 7: Experimental Agents and Therapeutic Drugs

Table of Drugs and Agents Used in Live AnimalsList all drugs or agents that will be administered to live animals in Table 7.A.

Drug/Agent: o Experimental agents include placebos (saline, vehicles), viruses and other biological agents, tumor cells,

stem cells, gene markers, tracers, radioisotopes, etc.o Therapeutic drugs include anesthetics, analgesics, tranquilizers, antibiotics, fluids and other agents such as

antimicrobials and paralytics.o Other study agents include saline, vehicles, disinfecting agents (betadine, alcohol swabs), ophthalmic

ointment, depilatory cream, etc.o Do not list euthanasia drugs in this section (euthanasia drugs should be identified in Section 8)o Rodent biological materials, including cell lines, may require testing for pathogens prior to use.

Purpose: Specify the purpose of the drug/agent in the study. Dose Range: Express dosages as a range and as mg/kg of body mass, wherever possible. Route: Indicate the routes of administration. Duration : Specify how long the drug or agent will be administered (e.g., 3 days, 2 weeks, etc>) Frequency : Specify how often it will be administered in each animal in a 24 hour period(e.g., every 8 hours)

Please list all agents in Table 7.A on the following page

Table 7.A - Experimental Agents and Therapeutic Drugs

Double-click on list below to enter study drug/agent informationTo exit table, click outside of the table

Section 8: Euthanasia Methods

A. Method of Euthanasia

Anesthetic Overdose

List the anesthetic agent(s) that will be used to euthanize animals. Indicate the dose and administration route.

Species Agent Dose(mg/kg)

Route of Administration

Physical Method

Place an X in any checkboxes that apply and describe the method in the following blank section below.

Decapitation:

Cervical Dislocation:

Cardiac Perfusion:

As described in Section 4.A.2

Other:

Will animals be anesthetized prior to physical euthanasia method being implemented?Yes *No

*If no, scientific justification must be provided:

B. Confirmation of Death in Animals Open chest inspection of the heart

Exsanguination (cutting a major blood vessel)

Physical method (indicate):

Other (describe below):

Section 9: Roles, Responsibilities and Experience of the Study Team

List below all study team members who will have contact with live animals. Describe each person’s specific role and responsibility on the project, including the procedures they will perform. Indicate who will be responsible for the daily care and monitoring of the animals. Provide a description of their qualifications, level of training and expertise. If a study team member does not have relevant experience or training for a particular species or procedure

they will perform, describe how they will be trained.

Principal Investogator:

Research Personnel:

Research Personnel:

Research Personnel: