September 20161

The Feinstein Institute for Medical Research

INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE (IACUC)

Animal Use Protocol

Administrative Section: (to be completed by IACUC office) Category E Protocol
Protocol Number: / Date of IACUC Review:

Please advise the IACUC Office upon submission of this protocol if your grant proposal or continuing grant funding requires IACUC approval by a certain date.

SECTION I

I.A. Title of Protocol:
I.B. Is this an Original Submission or Triennial Renewal?
Original submission / Triennial Renewal
I.C. If this protocol has been or will be reviewed for scientific merit by another committee, provide the name of the committee and date of review.
Note granting agency, submission date, title, and grant number (if available):
I.D. Funding Source:
Departmental Funds are available and will be used for this study / Funds not yet available / This protocol is part of a grant application / NIH/PHS funded
Note: If funds are available, please provide the grant fund number or cost center number below:

SECTION II –PRINCIPAL INVESTIGATOR

II.A. Principal Investigator (Include degree MD, PhD, other):
II.B. Status (F/T Faculty, Investigator, Associate Investigator, Assistant Investigator, other):
II.C. Department/Lab or Center of Excellence:
II.D. Site of Study (Feinstein Institute, LIJ, other):
II.E. Principal Investigator Contact Information:
Office Phone:
Email:
Cell or Alternate #:
II.F. Alternate Contact: This individual is authorized to make decisions on this study if the Principal Investigator cannot be reached.
Name:
Office Phone:
Email:
Cell or Alternate#:

SECTION III – EXPERIMENTAL SUMMARY IN LAY TERMS:

This summary should be in non-technical language and address issues that might be of concerns to persons with reservations about the use of animals in research

III.A. What species will be used?
III.B. How many animals are you requesting to use in the entire study (total number for 3 years)?
III.C. What is the overall objective(s)/hypothesis to be tested?
III.D. Harm vs. Benefit Analysis – answer this section in 3 parts:
  1. How could humans or animals benefit from your experiments?
  2. What are the potential adverse effects of this study on the animals?
  3. How do you, the Principal Investigator, weigh the potential adverse effects of this study against the potential benefits that may result from your research?

III.E. (LAY TERM SUMMARY)What will be done to the animals from start to finish? Please provide a simple summary that can be read by non-scientists. Brief description of procedures involving animals.
Detailed descriptions of any surgery should be provided in SECTION VI- SURGICAL RELATED MANIPULATIONS.
Detailed descriptions of all Non-Surgical Procedures should be provided in SECTION IX- NON-SURGICAL RELATED MANIPULATIONS.
III.F. Why must animals be used and why was this particular species selected?
III.G. Will any of the animals experience any pain or distress that cannot or will not be alleviated (Category E)?
Yes NoIf yes, justification required.
III.H. Will animals undergo more than one survival surgery?
Yes No- If yes, provide justification.
III.I. Does this protocol involve use of exogenous substances administered to animals? (e.g. recombinant or synthetic nucleic acid molecules, hazardous chemicals, exogenous substances, biologicals, infectious agents or bacteria)?
Yes No- If yes, complete Section X – Exogenous Material and Cells.
III.J. Does this protocol involve the use of radioactive material?
Yes No - If yes, consult with Radiation Safety Office 516-562-3895.
III.K. Will animals be irradiated?
Yes No - If yes, consult with Radiation Safety Office 516-562-3895.

SECTION IV - ANIMALS

IV.A. Species/Strain. (List all to be used)Complete section below. In addition to completing the table below, you may attach a table to this protocol application outlining the animals to be used per aim per experiment.
Species / Strain(s) / Gender / Age / Weight / Total #
Requested
Total:
IV.B. Number of animals that will be bred for experimental use (total 3 years):This number should be included in the total number of animals requested for the duration of the study.
IV.C. Total number of animals to be utilized in the entire study (total 3 years): The IACUC is mandated to ensure use of appropriate species, use of minimal number of animals necessary and that alternative approaches have been considered.
IV.D. Where will these animals be obtained from? (e.g. vendor, collaborator, other, etc.)
IV.E. Justification for the number of animals requested:Experimental design,Anticipated results,End of experiment
Include the number of experimental control groups and the number of animals per group for each experiment. Statisticians are available for consultation 516-562-0300.
IV.F. Will the animals require special conditions?
Yes No- If yes, describe (e.g. caging, temperature, light cycle, sanitation, diet, water, or isolation).
IV.G. Will restrictions be placed on any of the following in the conduct of this study:
a. the use of social housing- provide justification if any animals will be housed singly (if species is not considered “social”, then so note)
b. environmental enrichment or
c. the 8th edition Guide compliant cage size
Yes No - If yes, please describe the restrictions and provide scientific justification.
IV.H. Will the animals be removed from the Animal Facility during any phase of the protocol?
Yes No-If yes, specify where and for how long. Contact the Facility Director in advance to coordinate and obtain approval.

SECTION V – PERSONNEL - List all personnel working with live animals on this study.

All personnel listed must complete all pre-requisites and receive clearance from the Training Specialist.

V.A. - PERSONNEL- Who will perform experimental manipulations of live animals?

NAME / Procedures individual will be responsible to perform in this study / Office Phone, Cell, Email / If personnel require protocol specific (procedure) training, how will training be obtained?

SECTION VI- SURGICAL RELATED MANIPULATIONS NOT APPLICABLE

Requirements: Provide Justification for the following if applicable.

PRE-OPERATIVE CARE

VI.A. Withholding food
Yes No - If yes, specify how long.
VI.B. Withholding water
Yes No – If yes, specify how long.
VI.C. Antibiotics/Drugs
Yes No - If yes, provide:
Name / Dose / Route / Frequency

ANESTHESIA

VI.D. Will anesthetic(s) be used?
Yes No If yes, provide:
Name / Dose / Route / Frequency
VI.E. How will the depth of the anesthesia be monitored? (e.g. Absence of withdrawal reflex to toe pinch, monitoring of respiration)
VI.F. Will a paralytic agent be used?
Yes No - If yes, you must justify its purpose and explain how the level of anesthesia will be monitored while the animal is paralyzed.

SURGICAL PROCEDURES Not Applicable

REQUIREMENTS:Surgery will be performed under sterile or aseptic conditions. Agree

Sterile conditions are required for mammals (excludes rodents).

Aseptic conditions are required for rodents.

VI.G. Provide detailed descriptions of all surgical related manipulations:
VI.H. Will any animals on this protocol undergo a survival surgical procedure?
Yes No - If no, animal will be euthanized under anesthesia.
VI.I. Will animals on this protocol undergo a “major” surgical procedure? Note: Major surgery penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic function.
Examples of body cavities: cranial cavity, vertebral canal, thoracic cavity, abdominal cavity (including bladder and reproductive organs).
Yes No
VI.J. Will more than one survival surgical procedure be performed on any one animal? Note: Multiple major survival surgical procedures on a single animal are discouraged but may be permitted if scientifically justified and approved by the IACUC.
Yes No - If yes, please provide a justification.
VI.K. How will the skin be closed?Provide details below.
Suture material / Staples / Tissue adhesive / Other: describe

POST-SURGICAL CARE Not Applicable

VI.L. Describe post-surgical care and monitoring? (Include monitoring schedule ensuring evening and weekend coverage and removal of external sutures). Note: PI or personnel listed on this protocol are responsible for post-surgical care and monitoring animals once experiments have been initiated. If you require technical assistance, contact the Animal Facility Director.
VI.M. Describe precautions after surgery to prevent hypothermia and/or dehydration including duration (e.g. heating lamp, warming blanket, intravenous fluids, subcutaneous fluids).
VI.N. Criteria when animals can be returned to their housing (e.g. sternal recumbency, other; specify)

POSTOPERATIVE ANALGESIC(Consult with veterinarian required for category E studies) Not Applicable

VI.O. Will analgesic(s) be given to alleviate pain? Yes No
If Yes, please provide:
Analgesic Drug / Dose / Route / Frequency
If No, please justify:

SECTION VII- MINIMIZATION OF PAIN AND DISTRESS

USDA Pain Category / # of animals
Category B: / Category B animals are those that are being "bred, conditioned, or held for use in teaching, testing, experiments, research, or surgery but not yet used for such purposes." These animals have not been used for any research procedure, however minor. Category B is the place to put breeders and other animals that are not undergoing any experimental procedures.
Category C: / Category C animals are not subjected to procedures that involve pain or distress or would require the use of pain-relieving drugs. Routine procedures such as injections and blood sampling from veins that produce only mild, transient pain or discomfort are reported in this category. Another example of category C procedures is an observational study of animal behavior. Animals that are euthanized before tissue collection or other manipulations are also commonly placed in this category, if no other procedures performed that put them in a higher pain/distress category.
Category D: / Category D animals are those subjected to potentially painful procedures for which anesthetics, analgesics, or tranquilizers will be used. The important concept is that animals are given appropriate anesthesia and/or pain relief to limit their pain and distress as much as possible. Examples of category D procedures are surgery conducted with appropriate anesthesia and postoperative analgesia; Rodent retro orbital eye bleeding performed under anesthesia; Primate tattooing performed for identification under anesthesia; Removal of a small tumor under local or general anesthesia, and use of analgesia after an animal's skin is exposed to ultraviolet light to cause a "sunburn"; Terminal exsanguination (euthanasia by removal of blood) under anesthesia.
Category E: / Category E animals are those that are subjected to painful or stressful procedures without the use of anesthetics, analgesics, or tranquilizers. Withholding of anesthetics, analgesics, or tranquilizers can only be allowed if it is scientifically justified in writing and approved by the IACUC. Examples of category E procedures are lethal dose studies (e.g. LD50 studies) that allow animals to die without intervention, pain studies that would not be possible if pain-relieving agents were administered, and psychological conditioning experiments that involve painful stimuli such as a noxious electrical shock that cannot immediately be avoided by an animal.

SECTION VIII- EUTHANASIA AND STRESS REDUCTION

Stress Reduction

VIII.A. Is it expected this study will induce significant stress in the animals?
Yes No - If yes, detail how the stress be reduced or minimized? Indicate how long animals will be allowed to experience the stress, measures used to identify the stress, when and how you will intervene to address it.

Euthanasia

Acceptable Methods include CO2 asphyxiation, barbiturate overdose, and exsanguination under anesthesia.

VIIIB. Method of Euthanasia. Note: Must comply with 2013 AVMA Guidelines on Euthanasia.
VIII.C. Indicate specific criteria which will be used to identify moribund or distressed animals requiring early euthanasia.Note: Moribund is when an animal is in a dying state; at the point of death.
VIII.D. Will a physical method of euthanasia (e.g. decapitation, cervical dislocation, microwave irradiation of brain, electrocution – excludes CO2 asphyxiation) be used?
Yes No - If yes, you must provide a clear scientific justification for the method of termination.

SECTION IX- NON-SURGICAL RELATED MANIPULATIONS Not Applicable

Requirements: Provide Justification if applicable.

IX.A. List all non-surgical manipulations to be used. Please use this section to detail all procedures that will be performed on living animals.(Examples include but are not limited to tail snips, eye bleeds, ear notching, oral gavage and injections.)
IX.B. Anesthesia– Will you be using anesthetic agents for anynon-surgical procedure in this protocol?
Yes No - If yes, provide:Name, Dose, Route, Frequency for each anesthetic agent
Name / Dose / Route / Frequency
IX.C. Analgesics
Yes No - If yes, provide:
Name / Dose / Route / Frequency
IX.D. Antibiotics
Yes No - If yes, provide:
Name / Dose / Route / Frequency
IX.E. Extended Restraint
Yes No – If yes, provide:
Method - Chemical or Physical
(e.g. restraint device or cage) / Duration
IX.F. Behavioral Conditioning
Yes No – If yes, provide:
Purpose / Describe criteria for the monitoring
the condition of the animals
IX.G. Nutrition, Hydration and Dietary Manipulation
Yes No – If yes provide:
Purpose / Method / Duration

SECTION X- EXOGENOUS MATERIALS, RECOMBINANT DNA, CELLSNot Applicable

X.A. RECOMBINANT DNA

Does this protocol involve the use of Recombinant and Synthetic Nucleic Acid Molecules?
Yes No – If yes, provide IBC approval as an attachment to your submission.
IBC Registration #:
Date of Approval:
Registered Material Proposed for use in this Animal Study:
X.B.1. CHEMICAL GRADE EXOGENOUS MATERIAL(S)- Provide SDS sheet for each exogenous material, if available. Please submit a few references, if available, if the substances listed below have been used in this species before.
CHEMICAL GRADE
agent / List
(Include Dose, Frequency, Route of Administration and volume) / Liquid (L), Solid (S), Vapor (V) / Hazard Type (SDS Sec 3) / SDS Attached
Chemicals
Yes No
Radioisotopes
Yes No
Infectious Agents
Yes No
Antibodies
Yes No
Biological Compounds
Yes No
Other:
Yes No
*Provide justification for all chemical grade substances listed above.
*Describe how purity and sterility will be ensured for the chemical grade substances listed above.
X.B.2. PHARMACEUTICAL GRADE EXOGENOUS MATERIAL(S)- Provide SDS sheet for each exogenous material. If not available, please indicate such. Please provide a few references, if available, if the substances listed below have been used in this species before. A Pharmaceutical grade compound is a drug, biologic, or reagent that is approved by theFood and Drug Administration (FDA) or for which a chemical purity standard has been established by the United States Pharmacopeia-National Formulary (USP-NF), or British Pharmacopeia (BP).
PHARMACEUTICAL GRADE
agent / List
(Include Dose, Frequency, Route of Administration and volume) / Liquid (L), Solid (S), Vapor (V) / Hazard Type (SDS Sec 3) / SDS Attached
Chemicals
Yes No
Radioisotopes
Yes No
Infectious Agents
Yes No
Antibodies
Yes No
Biological Compounds
Yes No
Other:
Yes No
X.C. Will the animals experience changes or any Physical or Physiological Impairment as a result of the administration of exogenous material(s)?
Yes No - If yes, please justify:
X.D. Location where the exogenous material(s) will be used:
I understand that using biohazardous agents in animals will require the use of the BSL-2 suite. I agree to inform the CCP of the use of these agents so housing of animals in the BSL2 room use can be scheduled, and proper safety signs posted.
I understand that the use of hazardous exogenous substances in live animals requires the completion of the CCP Hazardous Exogenous Substances form (available through the CCP). This form must be approved by the Safety Office PRIOR to initiating any hazardous work in animals.
NOT APPLICABLE
CCP (Center for Comparative Physiology) / PI Lab (Building/Room): / Other area:
X.E. Indicate route of administration of all exogenous material(s):
Intraperitoneal / Intramuscular / Intravenous / Subcutaneous / Intracerebral
Orally / Intranasal / Inhalation / Gastric lavage / Other (list):
X.F. Does the agent occur naturally in the species used for the studies?
Yes No
X.G. Describe the expected time course of the experiments:
Number of experiments:
Duration of experiments:
Duration of potential exposure to agent(s):
X.H. Describe the personal protective equipment (PPE) that will be required for personnel working with animals that are exposed to the exogenous material(s):
Gloves / Jumpsuits / Masks / Eye Protections / Other (list):
X.I. Describe the method of disposal and decontamination of PPE:
X.J. Describe the protective measures that will be used with animals that are exposed to exogenous material(s).
Note: Standard ABSL (Animal Biosafety Level) will be performed by CCP personnel unless otherwise requested.
Cage changing / Standard for designated ABSL1-2 / Other (describe):
Bedding disposal / Standard for designated ABSL 1-2 / Other (describe):
Carcass disposal / Standard for designated ABSL 1-2 / Other (describe):
Additional measures not listed:

CELLSNot Applicable

X.K. Does this protocol involve the use of cells:
Yes No - If yes, provide:
Name and Type of Cell / Description of Cell Type / Primary or Cell Lines / Source
(Where obtained)
X.L. If source is cell lines, provide:
Prior Culture Conditions (describe): / Have these cells been RADIL tested?
Yes No –
If yes, attach documentation to this application.

Methods of Disposal for chemicals: Regular garbage, Red Bag, Yellow Bag, Chemical Waste pick-up

Note: All stock vials or unused agents must be discarded through the regular chemical waste disposal procedure. At theFeinstein Institute, please contact the Safety Officeat (516) 562–1050 and at LIJ contact Safety at (718) 470-4804.

Red Bag – (Biohazard) / Yellow Bag (Chemotherapy Drugs)
ALL ANIMAL CARCASSES. Bedding from animals treated with, all agents (biological, chemical, unknowns, proprietary substances, etc.) EXCEPT CHEMOTHERAPEUTIC DRUGS. / Chemotherapeutic Drug Waste ONLY. Contaminated bedding, gloves, paper, gowns, etc. with trace quantities of chemotherapeutic drugs. At the end of the day, all bags must be labeled “CHEMO WASTE” and the investigators must contact: Environmental Services (at NSUH (516) 562–2160 and at LIJ (718)470-7427, for removal of waste.

SECTION XI- ANTIBODY PRODUCTION (poly & monoclonal) Not Applicable

XI.A. Species:
XI.B. Antigen:
XI.C. Did you develop or isolate the antigen yourself?
Yes No- If no, indicate the source of the antigen and attach approval for use documentation to this application.
XI.D. Adjuvant (initial injection):
XI.E. Adjuvant (booster injections):
XI.F. Route of administration:
XI.G. Is Sedation used?
Yes No - If yes, provide:
Drug Name / When will sedation be used? / Dose / Route
XI.H. Will custom-made antibodies be obtained from another Institution or commercial vendor?
Yes No - If yes, provide name, PHS Assurance Number, and USDA Registration Number of supplier.

SECTION XII- TUMOR STUDIES (including Ascites) Not Applicable