Michigan State UniversityInstitutional Animal Care and Use Committee

APPLICATION TO USE VERTEBRATE ANIMALS IN RESEARCH, TEACHING, OR TESTING Revision date: August 2017

DATE RECEIVED: APPLICATION NUMBER: ROUND: AUF INACTIVE DATE:

DATE APPROVED:

DATE EXPIRES: PHS/NSF FUNDED

DOD FUNDED AGRICULTURE

USDA AMENDMENT DATE: ROUND:

AMENDMENT APPROVAL

IMPORTANT -- ALLOW 4-6 WEEKS FOR APPROVALINCOMPLETE FORMS MAY CAUSE REVIEW TO GO INTO THE NEXT REVIEW CYCLE DELAYING APPROVAL AN

ADDITIONAL 4-6 WEEKS

THIS FORM MUST BE SUBMITTED AS A WORD FILE VIA EMAIL TO: iacuc@msu.edu

ABBREVIATIONS AND ACRONYMS SHOULD BE SPELLED OUT AND EXPLAINED THE FIRST TIME THEY ARE USED.

Approval is for a maximum of three (3) years from the date of approval. Continuation of the approved animal usage beyond three years requires renewal of the AUF and complete IACUC review.

PRINCIPAL INVESTIGATOR(As designated by Contracts & Grants)

LAST NAME: FIRST NAME: MIDDLE INITIAL: ZPID NUMBER:

ACADEMIC RANK / TITLE: PRIMARY COLLEGE:

DEPARTMENT: CAMPUS TELEPHONE NUMBER:

MAILING ADDRESS: FAX NUMBER:

NET ID: CELL NUMBER:

CO-PRINCIPAL INVESTIGATOR(Regular / Fixed-term / Visiting / Adjunct / Clinical Faculty, Research Associate, Specialists, Postdoctoral Fellows)

LAST NAME: FIRST NAME: MIDDLE INITIAL: ZPID / STUDENT NUMBER:

ACADEMIC RANK / TITLE: PRIMARY COLLEGE:

DEPARTMENT: CAMPUS TELEPHONE NUMBER:

MAILING ADDRESS: FAX NUMBER:

NET ID: CELL NUMBER:

August 2017 Page 1

PRINCIPAL INVESTIGATOR'S ASSURANCE

I agree to abide by all federal and state regulations, Michigan State University (MSU) policies and Institutional Animal Care and Use Committee (IACUC) guidelines concerning the use of animals.

I agree that all use of vertebrate animals will be covered by an Animal Use Form (AUF) that has been reviewed and approved by the MSU IACUC and that IACUC approval must be obtained before ordering animals and/or performing any animal procedures described in this form.

I will promptly notify the Attending Veterinarian or designee regarding any unexpected study results that negatively impact the animals, including any unanticipated pain or distress and/or morbidity or mortality.

I agree that any proposed changes to this protocol will be requested by sending an amendment form, an amended AUF, and any related SOPs to the IACUC outlining the changes. IACUC approval must be obtained before performing the revised animal procedures described therein. I will maintain appropriate animal records (e.g., census, health, veterinary care, euthanasia, surgery, diagnostic, anesthesia, etc.).

I will do everything within my power to safeguard the health and well-being of each animal under this protocol.

I accept responsibility that all personnel working on this project are aware of and will follow the approved procedures outlined in this form. I assure personnel are adequately trained and have demonstrated competence in the animal procedures.

I understand that approval of projects is for a maximum of three years from the date of approval. I understand that the IACUC can call for a complete re-review of the project as needed.

By submitting this form, I agree to protocol-related activities including post approval monitoring, equipment maintenance, and communications with representatives of the IACUC at least annually.

PRINCIPAL INVESTIGATOR SIGNATURE DATE

CO-PRINCIPAL INVESTIGATOR SIGNATURE DATE

August 2017 Page 2

PROJECT/COURSE TITLE AND FUNDING AGENCY(if you have more than one project to put on this form, please copy & paste this page as many times as needed)

Title of Study (this has to match the grant title):

This is a renewal of a previously approved study. AUF #

Funding Source

PHS / NIH / NSF DOD / VA Other Federal Funding:

Internal / University Other (includes state funding, foundations, companies, and other organizations)

Contracts & Grants Application or Account Number:

Scientific merit of this project assessed by: (Funding agency, name of department representative, etc.)

Funding agency review Internally reviewed by:

Is this project a Sub-Contract from another Institution?

No Yes -- provide the name of the institution:

Will this project have Sub-Contracts to another Institution(s) involving live animal work?

No Yes -- provide the name of the institution:

List the animals requested for use, including the Use Category for each, and the number that will be used over the three-year period in the column corresponding to the source of animals. WHEN FILING AN AMENDMENT ADDING ADDITIONAL ANIMALS, DO NOT ADD TO THE EXISTING NUMBER – USE A NEW LINE.

Common name(do not list by strain)

Use category1

(one per line)

Number of Animals Requested Over a Three Year Period by SourceACCOUNT FOR ALL ANIMALS SUPPORTED BY THIS AUF

Total NumberNumber

transferred from another AUF

Number purchased or received from

another institution

Estimated number produced In-

house2

(include born but not used)

Other (Client-owned,

donated, wildlife, etc.)

1 Definition of Use Categories :Category B: Animals that will be bred or purchased for breeding, but not used for experiments. This includes breeders, offspring that cannot be used because of improper genotype or sex and any other animals that will not participate in the research studies. NOTE: animals that have been genotyped are NOT considered Category B.Category C: Animals used in research, experiments, or tests which involve no pain or distress or only momentary or slight pain or distress that WOULD NOT REQUIRE anesthetic, analgesic or tranquilizing agents (examples: s.c., i.m., i.p. or percutaneous i.v. injection, a brief period of restraint, tissue harvesting after euthanasia has been performed).Category D: Animals used in research, experiments, or tests where appropriate anesthetic, analgesic, or tranquilizing agents are used to avoid pain or distress (examples: major and minor surgery, tissue or organ collection prior to euthanasia, retro-orbital blood collection, prolonged restraint accompanied by tranquilizers or sedatives). Animals used in research, experiments, or tests which, if they experience pain or distress cannot be treated with an anesthetic, analgesic or tranquilizer, but the agent or procedure producing the pain/distress is immediately discontinued or the animal is euthanized to prevent pain and/or suffering.Category E: Animals used in research, experiments, or tests involving pain or distress in which the use of appropriate anesthetic, analgesic, tranquilizing agents or other treatments that would relieve the pain or distress would have adversely affected the procedures, results, or interpretation of the teaching, research, experiments, surgery, or tests (examples: studies which allow endpoints that are painful or stressful, addictive drug withdrawals without treatment, pain research, noxious stimulation). IF YOU LIST ANIMALS IN THIS CATEGORY YOU MUST PROVIDE A DETAILED JUSTIFICATION (see Section E)

2For protocols involving generation of offspring, list the number of breeders required and the expected offspring from all breeding. All animals born in-house must be included in this table, including any excess or unsuitable animals that may not be used for experiments; if you are breeding rodents, complete Section B.

August 2017 Page 3

VETERINARY CARE

A. Veterinary care will be provided by

Campus Animal Resources MSU Farms Veterinarian

College of Veterinary Medicine NA (Wildlife Field Study)

Van Andel Research Institute Other:

Private Veterinary Practitioner (provide name and contact information):

ANIMAL CAREPlease note that animals must be observed daily and health records must be maintained and readily available

B. Daily animal care will be provided by

Campus Animal Resources University Farm Manager

College of Veterinary Medicine Principal Investigator & Staff

Van Andel Research Institute NA (Wildlife Field Study)

Other:

ANIMAL HOUSING

C. Housing location:

Campus Animal Resources NA (Wildlife Field Study)

College of Veterinary Medicine Van Andel Research Institute

University Farms – If your animals will be housed in an Animal Science farm an ANS Animal Use Form needs to be approved by the Department of Animal Science (http://www.ans.msu.edu/internal_information/electronic_forms_1)

Other:

August 2017 Page 4

PROCEDURES

D. Please check all boxes below that best describe what will be performed on animals to be used in this application.

PROCEDURE/STUDY AREA

Animal Identification and/or Genotyping Methods 02

Antibody Production (complete Section I) 05

Behavioral 82

Biological Material / Recombinant DNA / Controlled Substances (complete Section D) 90

Breeding (complete Section B for rodents only) 09

Exceptions to Guidelines (complete Section J) 93

Food/water restriction (excluding surgical) (complete Section G) 20

Hazardous Chemicals (Biological, Chemical, Radioactive) (complete Section D) 91

Gaseous Anesthetic (complete Section D) 74

Induced experimental infectious disease 22

Induced experimental non-infectious disease 57

Injections or inoculations 83

Injury/trauma 26

Non-Invasive Procedure 92

Nutritional studies 51

Neuromuscular Blockers / Paralytic Agents (complete Question 24) 35

Obesity (experimental) 32

Organ/system failure/dysfunction experimentally induced 34

Unrelieved Pain & Distress (complete Section E) 31

Pharmacological studies 50Restraint of a conscious animal longer than 3 hours (complete Section F)(Wildlife Field Studies complete Section C) 38

Sample collection from live animals (blood, urine, tissue, etc.) (answer Q 25) 39

Sample collection (blood, urine, tissue, etc.) post euthanasia 99

Surgical procedure, non-survival excluding Perfusion & Exsanguination (complete Section A) 44

Surgical procedure, survival (complete Section A) 45

Surgical procedure, multiple major survival (complete Section A) 70

Teaching 61

Trapping/capture of wildlife (complete Section C) 13

Tumor growth experimentally induced / implantation 48

Weight loss 49

August 2017 Page 5

PERSONNEL

Please list in the table below all personnel who will work with live vertebrate animals or animal tissues as part of this Animal Use Form. Include Principal Investigator, co-Principal Investigator(s), Post-Docs, students, lab technicians, farm personnel, etc. The PI is responsible for ensuring that their employees and students are adequately trained in the specifics of their tasks. All trainings must be documented.

Mandatory trainings (final approval of your protocol will be withheld until training has been completed).

• IACUC Tutorial and IACUC Species Specific Tutorial (http://ora.msu.edu/train) • Biosafety Principles for Animal Users (www.ehs.msu.edu) • Medical Questionnaire (http://occhealth.msu.edu/resources/animal-contact)

Name, Title Phone MSU Net Id

Relevant Degree

orCertification

If multiple species, state with which

species personnel will be working

Man

dato

ry T

rain

ings

C

ompl

eted

(see

abo

ve)

PROCEDURES TO BE PERFORMED

Non

-Inva

sive

Act

ivity

Ane

sthe

sia

Eut

hana

sia

Sam

ple

Col

lect

ion

Sur

gery

Sur

gica

l Ass

ista

nce

Ass

ess

Hum

ane

End

poin

ts

Other

Dat

e of

f Pro

ject

(Offi

ce U

se)

i.e. John Doe, Professor 517-432-9999 Doej DVM cat, mouse x x x x x Husbandry

An IACUC species specific training module is not available in SABA; personnel will be trained by the Principal Investigator.

E. How have you determined that the personnel listed above are competent in performing anesthesia, euthanasia, surgery, and/or assessing humane endpoints?

August 2017 Page F

Michigan State UniversityInstitutional Animal Care and Use Committee

Animal Use Form Revision date: August 2017

DATE APPROVED: APPLICATION NUMBER: ANIMALS APPROVED: QTY

CLASSIFICATION OF THIS PROJECTFor clinical trial research, STOP and complete the AUF for Clinical Research

Biomedical Agriculture Teaching / Instruction Testing Wildlife / Field Study

Other:

LAY SUMMARYIt is NOT acceptable to use a section from your grant application, using highly technical terms. This information may

be used for press releases and/or responses to Freedom of Information Act (FOIA) requests.

1. The lay summary must be written in a manner in which the non-scientific members of the committee have an understanding of the relevance to human or animal health, the advancement of knowledge, or the good of society. A paragraph will generally suffice.

August 2017 Page 1

RATIONALE FOR ANIMAL USE / HARM-BENEFIT ANALYSIS

2. Please list the alternatives to the use of live animals that you have considered (e.g., the availability or appropriateness of the use of less invasive procedures, isolated organ preparation, computer simulation, and cell or tissue culture).

3. After considering the alternatives listed above, why do you think it is necessary to use live animals?

4. Justify the appropriateness of the species selected, rather than another or less sentient species.

5. Animal welfare regulations do not allow unnecessary duplication of previous experiments.

Does the proposed research duplicate previous work?

Yes No NA – This is a teaching activity involving different student groups

If yes, explain why it is scientifically necessary to replicate the experiment:

VETERINARY CONSULTATION

6. Are you a new PI submitting your initial Animal Use Form to the MSU IACUC?

No Yes -- complete a consult with a Campus Animal Resources (CAR) veterinarian

Are your animals classified as Category D or E?

No Yes -- complete a veterinary consult*

List the name of the veterinarian you have consulted regarding the planning of this project. You may contact:

• Campus Animal Resources (CAR) veterinarian at (517) 353-5064 (CARvets@msu.edu)• MSU Farms Veterinarian (517) 719-6165 (MSUAV@MSU.EDU)• MSU faculty member who is a graduate of a college of veterinary medicine• Veterinarian with expertise in the species being used

*With the exception of the Campus Animal Resources (CAR) veterinarians, to avoid any conflict of interest, personnel associated with this project cannot serve as the consulting veterinarian for the purposes of this review.

Name: Affiliation: Date:

August 2017 Page 2

PROCEDURES WITH THE POTENTIAL TO CAUSE PAIN AND/OR DISTRESS(Category D or E)

7. With respect to the procedures proposed that have the potential to result in pain and/or distress (e.g. surgery, anxiety/depression models, etc.), how have you determined that there are no alternative procedures that could be used instead?

USDA LITERATURE SEARCH

A literature search is required by the USDA for all covered species used in biomedical research, testing, and/or education activities which include the potential for pain and/or distress

(Category D or E)

Not USDA Covered -- A literature search is not required for:

• birds bred for use in research• rats of the genus Rattus for use in research• mice of the genus Mus bred for use in research• cold-blooded animals• farm animals used or intended for use as food or fiber• livestock or poultry used or intended for use for improving animal nutrition, breeding, management, or production

efficiency, or for improving the quality of food or fiber

8. As the USDA believes that literature searches remain the most effective and efficient method for demonstrating compliance with the requirement to consider alternatives to painful/distressful procedures, please provide the USDA required information in the table below. https://nal.usda.gov/sites/default/files/altwksht_march15.pdf

If you need help developing a computer search, contact: Andrea Kepsel, Health Sciences Educational Technology Librarian (akepsel@msu.edu) or (517) 884-0898

DATABASE NAMEmust search

two databases

DATE SEARCH PERFORMED

YEARS COVERED IN SEARCH

KEYWORDS/SEARCH STRATEGYmust include the words

“alternatives to pain & distress”

August 2017 Page 3

EXCEPTIONS

9. Does this AUF necessitate an exception* to the guidelines listed below?

YES – Please complete Section J (http://animalcare.msu.edu/iacuc-forms)

*Examples of exceptions:

Temperature / Humidity Sanitation / Waste Air Quality / VentilationSpace Requirements Disposal WaterFeed Noise Lighting IntensityBedding Acclimation

• Guide for the Care and Use of Laboratory Animalshttp://grants.nih.gov/grants/olaw/Guide-for-the-Care-and-Use-of-Laboratory-Animals.pdf

• Animal Welfare Act and Animal Welfare Regulationshttp://www.aphis.usda.gov/animal_welfare/downloads/Animal%20Care%20Blue%20Book%20-%202013%20-%20FINAL.pdf

• Guide for the Care & Use of Agricultural Animals in Research and Teachinghttps://www.adsa.org/Publications/FASS2010AgGuide.aspx

• MSU IACUC Guidelineshttp://animalcare.msu.edu/guides

August 2017 Page 4

NO

ENRICHMENT & SOCIAL HOUSING Please review CAR2: Enrichment Guidelines (http://animalcare.msu.edu/guides)

10. The standard practice at Michigan State University is to provide enrichment for the animals. CAR2: Enrichment Guidelines (http://animalcare.msu.edu/guides)

Does your project require an exception to this practice?

Yes No

If yes, provide scientific justification as to why animals cannot receive enrichment:

11. The primary emphasis of the enrichment program at Michigan State University is to house social species in compatible pairs or groups. Institutionally accepted exceptions are as follows:

• It is a normal agricultural husbandry procedure• The animal is incompatible as documented in its record• The animal is recovering from surgery, up to 72 hours • When multiple females are housed with males - the pregnant female may be singly housed when visibly

pregnant• When male rodents are used in multiple cages for breeding (generally they cannot be integrated back into

group housing)

Does your project require an exception to social housing not covered above?

Yes No

If yes, scientifically justify why animals will not be socially housed:

12. Will animals be maintained outside approved housing areas for more than 12 hours?

Yes No

If yes, provide scientific justification and indicate where the animals will be maintained:

August 2017 Page 5

USE OF CELL LINES IN ANIMALS

13. Will you be using cell lines in your research?

Yes No

If yes, what is the origin of the cells (human, animal species)?

Has the cell line been verified to be free of murine pathogens?

Yes No – Contact Campus Animal Resources (CARvets@msu.edu)

USE OF GENETICALLY MODIFIED ANIMALS

14. Does this project involve genetically modified animals?

Yes No

If yes, to assist the veterinary staff, list any known phenotypic variations of the GMA/Strain(s) used within this AUF that could potentially affect animal health, well-being or longevity.

USE OF CUSTOM-MADE ANTIBODIES IN LIVE ANIMALSPlease review: IG030: Antibody Production for PHS Funded Research (http://animalcare.msu.edu/guides)

15. PHS policy states: Antibodies are considered customized if produced using antigen(s) provided by or at the request of the investigator (i.e., not purchased off-the-shelf). An organization producing custom antibodies for an awardee must have or obtain an Assurance, or be included as a component of the awardee’s Assurance.

Will you be purchasing custom-made antibodies?

Yes No

If yes, provide the PHS assurance number of the organization producing the antibodies:

August 2017 Page 6

PROJECT DESCRIPTION

16. If your protocol will utilize standard operating procedures (SOP), please review the instructions on the use of SOPs http://animalcare.msu.edu/iacuc-forms

List the approved SOPs in this research. Append the approved SOP to this AUF.

APPROVED SOP NUMBER: TITLE OF APPROVED SOP: LAST NAME OF PI LISTED ON SOP:

17. Identify the Procedure / Experiment / Activity (if you have more than one procedure/experiment/activity, please copy and paste questions 17, 18, & 19, as many times as needed).

Procedure/Experiment/Activity:

Species: Number of animals: Use Category (for this procedure / experiment / activity):

Will any of these animals be used in another procedure/ experiment/activity within this AUF? If so, how many?

Species: Number of animals: Use Category: Will any of these animals be used in another procedure/ experiment/activity within this AUF? If so, how many?

In which building will the procedure be conducted? Room Number:

18. Provide scientific justification for how you determined the number of animals needed for this procedure/experiment/activity. For guidance see: Scientific Justification for the Number of Animals to be used in Research, Teaching or Testing (RD001). http://animalcare.msu.edu/iacuc-forms

19. Explain the experimental design of the above procedure/experiment/activity. Your target audience is an external inspector and/or faculty member from a discipline that may be unrelated to yours.

Include only the information that explains what is being done to the animals. Do not cut and paste from a grant proposal. Provide a timeline for the procedures proposed (from acquisition to final disposition) denoting the

approximate timing of all important manipulations For surgical procedures, please include the name of the surgery and where within the timeline the

surgeries will be performed; a detailed description of the surgery(s) is to be provided in Section A Please indicate where drugs and compounds will be used within individual procedures; in depth

details regarding drugs/paralytic agents are to be provided in questions 23 & 24 For sample collection from live animals, please indicate when samples will be procured; in depth

details regarding this is to be provided in question 25 If applicable, provide a brief overview (1or 2 sentences) of work done on tissue post euthanasia

ADVERSE CONSEQUENCES

August 2017 Page 7

Please review: IG001: Reporting Unexpected Adverse Events (http://animalcare.msu.edu/guides)

Please note: If any unanticipated adverse consequences not described below do occur during the course of the study, a complete description of these consequences and the steps taken to mitigate them must be submitted to the Attending Veterinarian or designee. Veterinary consultation must occur when pain and distress is beyond the level anticipated in the protocol description or when interventional control is not possible.

20. List and describe known adverse consequences that the animals could potentially experience as a direct result of the procedures described. Examples of adverse consequences might include post-operative complications, unkempt coat, ascites, difficulty rising, inflammation, self-trauma, weight loss, loss of appetite, paralysis, death, etc. These are consequences that you could reasonably predict based on your experience or knowledge of the literature.

21. What supportive care will be provided for animals experiencing adverse consequences?

22. For those animals experiencing adverse consequences, describe the criteria (i.e. clinical signs) that dictate when an animal will be euthanized (this is referred to as the humane endpoint).

August 2017 Page 8

DRUGS AND/OR AGENTSSurgical drugs should be listed in Section A

PIs are expected to use pharmaceutical-grade medications whenever they are available, even in acute procedures. Non-pharmaceutical-grade chemical compounds should only be used after specific review and approval by the IACUC.

Please review: IG005: Use of Non-Pharmaceutical Grade Compounds (http://animalcare.msu.edu/guides)

23. List drugs, compounds and/or agents used in this project except drugs and/or agents used in surgical procedures, which should be included in Section A.

This may include drug categories instead of specific drug names (e.g., classes of receptor agonists and antagonists; calcium blockers; diuretics; serotonin reuptake inhibitors).

The following will need to be listed by its actual name:• those that have potential adverse consequences• analgesics, anesthetics, and sedatives • antibiotics or other drugs used for therapeutic treatment• euthanasia agents • hazardous agents (e.g., chemical hazards, biologic hazards) that need the attention of EHS

Drugs that are not pharmaceutical grade should be identified as such. The use of non-pharmaceutical grade compounds must be scientifically justified.   In addition, if non-pharmaceutical grade compounds are to be used the PI should take precautions to maintain aseptic conditions and the drugs should be:

• of the highest purity available• adjusted to ensure a pH level as close to neutrality as possible• constituted in a sterile solution for administration to the animal• kept in a sterile, sealed container after constitution for a limited period of time• administered with sterile devices

√if not

pharmagrade

drug name(s)FILL IN BOTH route of

administrationfrequency of

administration

duration of treatment

(# of days or hours)volume dosages(range)

Scientifically justify use of non-pharmaceutical grade compounds:

August 2017 Page 9

24. List neuromuscular blockers / paralytics that will be used:

drug name(s)(generic only)

dosages(range)

route ofadministration

frequency ofdosages

duration of treatment(# of days or hours)

Scientifically justify why you need to use a paralytic:

SAMPLE COLLECTION(tissue and/or body fluids)

25. Complete the table below for any specimen collection from LIVE animals

type ofsamples collected

site ofsample collection

method ofsample collection

volumeof sample

number ofsamples

frequencyof collection

CARCASS / TISSUE / BODILY FLUIDS DISPOSAL

26. Does the Project/Experiment/Activity require the collection of animal carcasses, tissue, and/or bodily fluids?

Yes No

If yes, how will they be disposed?

August 2017 Page 10

FINAL DISPOSITION

27. Check all that could apply:

Euthanasia

Transfer to another PI with an approved AUF

Adoption (a record of adoption must be kept in the files)

Animals will remain at or be returned to the appropriate MSU managed facility

Client Owned/Privately Owned Animals will remain with their owners -- Q 28 & 29 do not need to be completed.

Wildlife release

Other (please explain):

EUTHANASIAConfirmation of death is required for all methods of euthanasia*

28. 1Even if your study does not involve euthanizing the animals you should provide an emergency method that you would use in the event of unanticipated injury or illness. Exceptions to this policy: client owned/privately owned animals or animals housed within an MSU managed facility.

Primary Method = CHEMICALNOTE: for inhalant/CO2 euthanasia, a physical method (options B through F) must be used to confirm death

√if not

pharmaceuticalgrade*

species(common name) agent dose (mg/kg)

or flow rateroute of

administrationmethod of death

confirmation*

[ ] I will use the CAR Facility Euthanex® Machine

* Scientifically justify use of non-pharmaceutical grade compounds being used for euthanasia (unless previously justified)

Primary Method = PHYSICALNOTE: for a physical method of euthanasia, a secondary method (options A through F) must be used to confirm death

species(common name) method of euthanasia method of death

confirmation*

*Confirmation of Death MethodsA = no heartbeat/corneal reflex B = decapitation C = bilateral pneumothorax

D = cervical dislocation E = exsanguination / perfusion F = Other (please state above)

29. Is the method of euthanasia recommended by the current AVMA Guidelines on Euthanasia (https://www.avma.org/KB/Policies/Documents/euthanasia.pdf)?

Yes No

If no, provide scientific justification:

August 2017 Page 11