Harvard Medical Area Page 1 of 46 Protocol # Revised 11/3/15 ANIMAL EXPERIMENTATION PROTOCOL Principal Investigator Harvard Appointment Title Department Address HMA phone FAX phone Home phone E-Mail Address Contact Person Contact Phone & Email BIDMC PI Check here: ☐ BIDMC investigators must send an approved copy of this protocol to BIDMC, East Campus, RN150D Protocol Title Funding Source Grant Number & Title Please attach the Vertebrate Animal Section for any new grants. Check one: New Protocol Three Year Review of Existing Protocol This proposal has been reviewed and approved by the HMA Standing Committee on
Transcript
Harvard Medical Area Page 1 of 35 Protocol # Revised 11/3/15
ANIMAL EXPERIMENTATION PROTOCOL
Principal Investigator
Harvard Appointment Title
Department
Address
HMA phone
FAX phone
Home phone
E-Mail Address
Contact Person
Contact Phone & Email
BIDMC PI Check here: ☐ BIDMC investigators must send an approved copy of this protocol to BIDMC, East Campus, RN150D
Protocol Title
Funding Source
Grant Number & TitlePlease attach the Vertebrate Animal Section for any new grants.
Check one:
New Protocol Three Year Review of Existing Protocol
This proposal has been reviewed and approved by the HMA Standing Committee on Animals
HMA Standing Committee on Animals Date:
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Table of Contents
Sections listed in BOLD must be completed for every protocol.
Sections F through N are to be completed only as they apply to your protocol.
HCCM’S MANDATORY RISK DISCLOSURE (MRD)
A. ANIMAL USAGE
B. PROCEDURES
C. PERSONNEL
D. HOUSING/PROCEDURE LOCATION
E. DISPOSITION OF ANIMALS
F. IMMUNIZATION
G. ASCITES PRODUCTION
H. TRANSGENIC/KNOCKOUT ANIMALS
I. ADMINISTERED SUBSTANCES
J. SURGERY
K. MULTIPLE SURVIVAL SURGERY
L. PAINFUL PROCEDURES
M. BEHAVIORAL CONTROLS, RESTRAINT, OR ANALYSIS OF BEHAVIOR
N. DIET AND OTHER ENVIRONMENTAL CHANGES
O. DRUG AND SURGICAL SUPPLY MANAGEMENT PLAN
P. ALTERNATIVES TO PAIN AND DISTRESS
Q. MINIMIZATION OF DISCOMFORT, PAIN AND DISTRESS
R. DUPLICATION OF RESULTS
S. PROTOCOL PERSONNEL SIGNATURE PAGE
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PI must print and sign below (mandatory; a proxy will not be accepted):
P.I. Name ___________________ P.I. Signature ________________________ Date __/__/__
**MUST CHECK (X) ALL THAT APPLY**Check: Procedures: Identify specific agent(s): Route(s):
☐ Chemical carcinogen(s)
☐ Toxic substance(s)
☐ Radioisotopelicense #:______________
☐Microbiological Agent(s)COMS/PIBC #: _______Provide Biosafety Level next to each agent
☐ Human or NHP cell line(s) or tissue(s)COMS/PIBC #: _______
☐ Non-human animal cell line(s) or tissue(s)
☐ Other (must specify)
☐
NONE OF THE ABOVEThis box must be checked if none of the above agents apply to your work described in this protocol.By checking this box & signing below you are emphatically stating that you are NOT using any of the above in research animals.
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Introduction
The Harvard Medical School Standing Committee on Animals was established in 1907 almost 60 years prior to the Animal Welfare Act. This Committee charter states: A Committee of at least five, which has the power to inspect the animal rooms at any time, to issue orders and regulations they may deem fit for the proper care and feeding of animals, to discharge any employee of the school who in any way treats animals with cruelty, to dispose of any animal which is in any way objectionable and to determine the sources from which animals are to be obtained. This Committee shall also have general power to stop any experimental work on animals that they may consider improper or unnecessary. This charter is still in effect today and the Committee, appointed by the Dean for Academic and Clinical Programs, continues to have the authority as described.
The Committee members today include scientists, veterinarians, and lay representatives. This Committee is committed to the continued use of animals in research within the constraints of humane treatment as viewed by society. To justify approval of procedures, the Committee must have adequate written information regarding the steps taken to assure humane treatment of animals. Present your material clearly and concisely. Lay members of the Committee must be able to understand this protocol.
The Animal Experimentation Protocol (AEP) MUST include a FLOW CHART AND a written description of the procedures outlined in Section B.2. Failure to comply will result in delay of approval for an additional month.
PIs are encouraged to send any new protocols and amendments as soon as the PI completes a reasonable draft.
Three-year renewals are due to the IACUC no later than 60 days prior to their expiration date in case the protocol needs to be re-reviewed at a subsequent full committee meeting. Otherwise there is risk of research interruption if any required modifications are not satisfactorily addressed prior to the protocol expiration date. Early submission helps ensure that there is no lapse in approval and that research continues uninterrupted.
Submission addressSubmissions and scanned signature pages should be sent electronically to: [email protected] (MS Word format only).
Accessing protocol formsCurrent forms and supplemental information can be found at: http://hms.harvard.edu/departments/hma-standing-committee-animals
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A. ANIMAL USAGE
1. Checklist – animal usage PER YEAR Place an “X” next to the procedures you will be performing. Enter the total number of animals in each category as well as the total number in multiple categories. Complete a separate ‘Checklist’ for each species. Complete a search for alternatives for each procedure in Category D and/or E
Species
# Used per year
CATEGORY B -Teaching, research, or tests that involve breeding or housing of animals only☐ Breeding colony☐ Housing ONLY (no other procedures)
Total number of animals to be used in Category B only
CATEGORY C -Teaching and/or research experiments that do not involve more than momentary pain and/or distress or require the use of pain relieving drugs.☐ Alert animals (behavioral observation or brief restraint)☐ Anesthetize and release for non-invasive procedures (e.g. imaging, bandage change)☐ Change in environmental parameters (diet, light cycle, room temperature, etc.)☐ Chemical carcinogens (e.g. BrdU)☐ Euthanasia with or without tissue/organ harvest☐ Forced exercise☐ Gavage☐ Irradiation☐ Microbiological agents☐ Non-surgical collection of body fluids (blood, urine, etc.)☐ Radioisotopes☐ Simple injections (IP, IV, etc.)☐ Tail biopsy for genotyping (for mice 21 days of age or younger)☐ Use as a parasitic host
Total number of animals to be used in Category C
Note: For all procedures in Categories D & E (below) a search must be completed in Section PCATEGORY D – Teaching and/or research experiments or tests that will involve pain and/or distress and for which appropriate anesthetic, analgesic or tranquilizing drugs will be used.☐ Anesthetize and release for non-surgical painful procedures (e.g. retro-orbital bleeds)☐ Antibody production: polyclonal (non-ascites, no footpad)☐ Burns☐ Controlled drugs or substances (as test material, e.g. cocaine)☐ Electric shock☐ Food or water deprivation other than pre-surgical fasting
☐ Introduction of illness (including the administration of toxins, microbiological agents, clinical transgenic phenotypes) WITH pain relief
☐ Lavage
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☐ Multiple MAJOR survival surgery☐ Multiple MINOR survival surgery☐ Non-survival surgery☐ Physical trauma☐ Survival surgery: MAJOR☐ Survival surgery: MINOR☐ Tail biopsy for genotyping (for mice over 21 days of age)☐ Toxic substances (e.g. streptozotocin, MPTP, cisplatin, doxycycline)☐ Transgenic mouse production (embryo manipulations to produce novel strains, etc.)☐ Tumor induction or implantation
Total number of animals to be used in Category D
CATEGORY E – Teaching and/or research experiments, surgery or tests that involve PAIN and/or DISTRESS, and for which appropriate anesthetic, analgesics or tranquilizing drugs are NOT used because they would adversely affect the results or interpretation of data.☐ Antibody production: Ascites☐ Death-as-an-endpoint or lethal dose studies*☐ Footpad injections (antibody production or microorganism)
☐ Induction of illness (including the administration of toxins, microbiological agents, clinical transgenic phenotypes) WITHOUT pain relief
☐ Noxious stimuli from which there is no escape☐ Pain study☐ Paralysis☐ Total body irradiation (without reconstitution)☐ Unusual or prolonged restraint (e.g. pie chamber, chair)☐ Withholding of post-operative analgesia for any surgical procedure
Total number of animals to be used in Category E*You must follow the IACUC Guidelines for ‘Death as an Endpoint’ or ‘Lethal Dose’ studies.
EXCEPTIONS to STANDARDS – Select any policy deviations that will be required in this protocol. Scientific justification must be included in the indicated section of the protocol.☐ Single housing of animals Section D.5☐ Withholding of enrichment from singly housed animals Section D.5.c☐ Euthanasia methods that are not consistent with the AVMA Guidelines Section E☐ Ascites collection Section G
☐ Non-pharmaceutical grade drugs and other compound suspensions when USP-grade drug or alternative is available.
Section I.7.c
☐ Withholding postoperative analgesia Section J.2.k☐ Food/water withholding or scheduling (not ad-lib) Section N☐ Smaller caging than the recommended space allotment (e.g.: CLAMS) Section D.4☐ Deviations from housing density standards Section D.6☐ Toe-clipping for identification purposes Section D.3.b.iii☐ Deviations from the 12/12 hour light/dark cycle Section N
☐ Any other exceptions to the standards of the PHS Policy, the Guide, or the AWA. - Please specify:
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2. JUSTIFICATION FOR THE USE OF ANIMALSFederal Assurances: You are required by federal law to provide the following justifications.
a. Why are animals needed for this study? (i.e.: Could the same information be obtained by experiments using tissue culture or computer models? If you are generating antibodies, could you use in vitro methods?)
b. Justify choice of each species:
c. Justify the number of animals to be used: (You must convince the Committee that the number of animals is appropriate to the work being proposed. Provide groups and statistical analysis to be used if relevant.)
d. Discuss your strategy to reduce the number of animals and to minimize pain and distress to your animals: (examples include: earlier euthanasia, decreasing the number of time points, changing the methods to those that are less invasive):
e. Breeding Colony users only: The estimated numbers of animals in breeding colonies must be provided and justified. These numbers must be also be tracked on a log sheet in the animal housing room.
i. Estimated number of breeding adults:
ii. Estimated number of offspring produced:
iii. Estimated total number of offspring euthanized without research use:All offspring euthanized without research use should be accounted for under Category B of the checklist.
iv. Criteria for offspring euthanized without research use:
f. NHP users only: Are the requested animals naïve in terms of experimental procedures? Yes ☐ No ☐
If NO, submit the approval for reuse summary from the Animal Allocation and Reuse Committee.
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B. PROCEDURES Briefly describe the procedures to be conducted on animals. State the aims and significance of your
experiments. Do not include in vitro work done on tissue taken from the animal, nor post-mortem procedures or analysis.
For 3-year reviews, incorporate all procedures listed in approved amendments in the past three years, if applicable. All amendments expire at the protocol 3-year renewal date.
Use language that can be readily understood by investigators and lay persons not familiar with your specific field of research and by non-scientists. Avoid or explain all acronyms.
Note: AEP submissions receive veterinary review prior to final submission to, and consideration by the IACUC. Veterinary pre-review and addressing veterinary comments do not guarantee Committee approval.
1. Aims and Significance: Provide a brief lay summary, no more than one page, describing your research. Do not include an excerpt from your grant proposal.
2. Procedures and manipulations: In lay terms, provide a short description of each animal procedure and manipulation (including changes in light cycle, alteration of feeding, etc. DO NOT include details of in vitro work). For SURGERY, state ONLY the name of the procedure in this section. Describe the surgery in detail (including anesthesia and site prep) only in Section J
3. FOR 3-YEAR REVIEWS:
a. For USDA-covered species (not mice, rats, or birds), describe any unexpected animal deaths in the course of your experiments:
b. Changes (briefly summarize any changes in the protocol in the past three years, i.e. species, numbers, procedures, etc.):
c. Progress Report (optional): Briefly describe your results from the past 3 years and explain how the present application continues this work:
d. List publications and meetings in the past three years (optional):
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FOR ALL (NEW and 3-YEAR Review) protocols
4. FLOWCHART - you must diagram your procedures in a flowchart
Present an overview of your experiments in a flowchart format (as opposed to a table, grid, or paragraph). Show the schedule for the animal procedures in your flow chart, including the time points when the animals
will be euthanized, if applicable. For guidelines on creating your flowchart click here. Excessively long flowcharts may be rejected.
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C. PERSONNEL
1. Will the principal investigator perform the procedure(s)? Yes ☐ No ☐
2. Personnel List:
Provide the name(s), title(s) and qualifications for each person who will work with the animals. The Principal Investigator must also be included.
Include the number of years and experience working with the species listed in Section A (Checklist) above.
If the person needs to be trained, indicate who will do the training. If this is a three year renewal protocol: The list below WILL REPLACE ALL CURRENT LAB
PERSONNEL. Any additional personnel must be added via a personnel form, after renewal. Members of the veterinary staff do not need to be listed.
Name Title/Job Classification
Qualification/ Experience
Phone Email Address
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
3. Personnel Training:
All personnel handling animals must be trained in their humane care and use. Records of personnel training must be maintained by the lab.
State the building and room number where training records for lab personnel will be stored:
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AALAS Learning Library TrainingClick here for the AALAS Learning Library FAQ.
D. HOUSING/PROCEDURE LOCATION
1. Where will your animals be housed?
Species Building (Circle or boldface one) Room #
WAB HSPH2 THORN HIM NRB SGMB MCP 65 Landsdowne
WAB HSPH2 THORN HIM NRB SGMB MCP 65 Landsdowne
For all research using rodents, indicate the level of biosecurity that you require. Specific-pathogen-free (SPF) facilities are free from infection with a defined panel of infectious agents. These areas ONLY house animals from approved vendors or those animals that have undergone HCCM quarantine. Animals in SPF facilities are maintained so as to prevent infection and undergo regular serologic surveillance to assure SPF status. Non-SPF facilities may harbor one or more of these pathogens.
Species SPF Non-SPF BL2/BL3
2. Location of your animals:a. Provide the area(s) where your animal procedures will be performed, including euthanasia.
Species Building Room # Planned procedures Time in this area**
**Animals may not remain in a lab for more than 12 hours, nor may they be housed in the lab overnight, without express approval from the IACUC.
b. Please check situations or locations to which you plan to transport animals:
Vendor-placed tag or tattoo *Toe Clipping Micro-chipping Other (Specify)
b. Describe the details of your method for identifying your animals:
i. Describe the procedure for identifying your animals (vendor-placed tags/tattoos excluded):
ii. Age of animal at time of ID procedure (note: Analgesia may be required under certain conditions):
iii. Scientific justification is required for toe-clipping. All other methods of identification must first be considered. State your justification here (you must contact the IACUC office to request an Exemption form):
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4. Will your animals be housed in the CLAMS metabolic caging system? Yes* ☐ No ☐*The use of CLAMS metabolic caging encompasses three deviations from standards: Cage size is smaller than standard, exemption from environmental enrichment, and social species housed individually. If you select “yes” here, you do not need to select each exemption individually.
a. Scientific justification for use of CLAMS metabolic caging system:
☐ The CLAMS system is required and used standardly for metabolic monitoring and activity testing. The mice will only be in the cages for a few days at a time (at most). When doing gas exchange measurements, it is advisable to keep the chamber volume as small as possible so as to have the system be as responsive to changes in the animal's metabolism. This involves a trade-off in floor area and/or height. Columbus Instruments elected to provide adequate floor space so that activity could be sensed within a reasonably sized space. The present chamber is approximately 2.6 liters in volume. When the chamber is configured for use in CLAMS, we add a floor that raises the animal above the waste material and affords easy access to the feeder system. Raising the walking surface does reduce the head space. Thus far, all clients that use the system (for a total of 1,400 or so installed cages) have not met with objections to the head space issue because the animals are in the system for a short period of time, i.e. a couple of days.
b. Or provide your own scientific justification:
5. Will your animals be socially housed (more than one animal per cage)? Yes ☐ No* ☐*The HMA IACUC requires social housing of all social animals. If ‘NO’, provide scientific justification for not socially housing the animals. Describe what will be done to replace this social contact with conspecifics.
a. Scientific justification for non-social housing of animals:
b. Methods for replacing socialization:
c. Will individually housed animals receive environmental enrichment?
d. If ‘NO’, provide scientific justification for withholding environmental enrichment:
6. Will your animals be housed in excess of housing density guidelines? Yes ☐ No* ☐
a. Scientific justification for housing density deviation:
E. DISPOSITION OF ANIMALS (Note: Any applicable endpoints noted below must be included in the flowchart in Section B)
1. Euthanasia Will you be euthanizing animals on this study? Yes ☐ No☐
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If you are using CO2 or any inhaled anesthetic (e.g. Isoflurane) as a means of euthanasia, you MUST also use a second physical means to ensure death (e.g. bilateral thoracotomy, exsanguination, or harvest of an essential organ, etc.).
Guillotining without anesthesia is not acceptable unless scientifically justified and approved by the Committee.
Cervical dislocation without anesthesia may ONLY be performed by personnel who have been properly trained.
Dry ice is NOT AN ACCEPTABLE METHOD of euthanasia under any circumstances.
Any deviation from the AVMA Guidelines on Euthanasia must be justified below:
If you answered ‘YES’ above, you must answer the following (copy and paste the table below for each additional species):
Species:
Method of euthanasia:
Secondary method of euthanasia if using inhalant overdose as primary method:
Justification for any deviation from AVMA guidelines:
Anesthetic/tranquilizing agent(s), if applicable:
Dosage:
Route of administration:
2. Transfer
a. INTERNAL TRANSFER – a transfer of animals to another protocol (either one of your own or that of another PI) within the jurisdiction of the HMA IACUC. NOTE: if your animals have had procedures performed, other than genotyping, the protocol on which the animals are being transferred to must be amended to include the animals that have undergone the prior procedure(s).
Will you be transferring any of your animals internally? Yes ☐ No ☐
If you answered ‘YES’ above, you must contact the HCCM to fill out the appropriate transfer form from the HCCM website.
b. EXTERNAL TRANSFER - a transfer of animals to another institution (outside of the HMA IACUC’s jurisdiction)
Will you be transferring any of your animals externally? Yes ☐ No ☐
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If receiving collaborator information is not yet available, answer “No” above and submit an External Transfer Form to the IACUC office when this information becomes available and at least one week prior to the animal transfer.
If you answered ‘YES’ above, you must complete the following (copy and paste for each collaborator):
i. Species:
ii. Receiving Investigator:
iii. Receiving Institution:
iv. Transport Method ☐ Walking
☐ Commercial shipper (list here):
vi. Principal Investigator ASSURANCES:
☐ I agree to ship ONLY to the Animal Resource Center at the Receiving Institution and NOT to an individual investigator or laboratory.
☐ I agree that the receiving Investigator has a current approved protocol for these animals.
☐ I agree to notify the receiving Investigator of any prior genotyping or identification procedures that have been performed on these animals.
☐ I agree to provide long-term gel-packs or equivalent if the animals will travel for more than 3 hours.
3. Dual-Assignment (USDA covered species only)
Will any of these animals be concurrently assigned to other protocols? Yes ☐ No ☐
Approval of a protocol that uses animals assigned to other protocols binds the principal investigator into an agreement to coordinate all animal sampling and procedures with the corresponding investigator to ensure compliance with all guidelines of federal agencies and the IACUC. Please see the Dual Assignment Policy for further information.
NOTE: Dual protocol assignment requires prior approval by the Animal Allocation and Re-use Committee (AARC) via the “Dual–Assignment Request Form.”
Protocol # Species # of animals Pain category (C,D,E)
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5. a. Are you performing footpad injections? Yes ☐ No ☐
b. If "YES", you must justify the use of footpad injections rather than other methods of immunization and you must provide assurance that the animals will be housed on soft bedding and have easy access to food and water.
NOTE: Footpad injections are allowed in ONE FOOT PER ANIMAL and are permitted only if justification is deemed sufficient by the IACUC.
Justification for footpad injection:
G. ASCITES PRODUCTION
1. Will the animals be used to generate ascites for any purpose? Yes ☐ No ☐
**Be advised that there is evidence that the ascites method of polyclonal or monoclonal antibody production causes discomfort, distress, and/or pain. Practical in vitro methods exist which can replace the ascites method in many experimental applications without compromising the aims of the study.
If “NO,” proceed to Section H
2. If “YES,” provide scientific justification for use of in vivo ascites production. Describe the method(s) you will employ to minimize discomfort, distress and pain and fill out the following table (NOTE: Animals MUST be checked at least once a day – INCLUDING WEEKENDS AND HOLIDAYS - when producing ascites fluid. Animals may be tapped a maximum of 3 times, with the fourth tap being a terminal tap followed by euthanasia.):
a. List methods (e.g. in vitro) found unsuitable (cost is not considered a suitable reason for exclusion):
b. Provide scientific justification for ascites production (cite references here):
c. Describe the methods that will be employed to avoid or minimize discomfort, distress and pain:
d. Where will the animals be housed?
e. How often will the animals be checked by laboratory personnel?Laboratory personnel must be listed in section C or added via New Personnel Form.
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H. TRANSGENIC/KNOCKOUT ANIMALS
1. Will your study involve transgenic or knockout animals? Yes ☐ No ☐
If "NO", proceed to Section I
2. If “YES”, complete the following:
a. Describe the phenotype
b. Describe any pain or distress associated with the phenotype expressed and your proposed procedure(s) to avoid or alleviate pain:
c. Describe your monitoring schedule.
d. Define the duration of survival after expression of the phenotype:
e. Describe the criteria you will use to determine pain and distress in the animals and your plan for monitoring the animals:
f. The construction of transgenic animals must be approved by the Committee on Microbiological Safety (COMS) or Partners Healthcare Institutional Biosafety Committee (PIBC).COMS or PIBC Registration #Contact the Biosafety Officer at 617-432-4727 (HMS/HSPH) or 617-964-8550 (NRB/HIM) or 800-825-5343 (BWH) for appropriate registration.g. If receiving animals from an outside source, identify the origin of the animals, or their COMS/PIBC Registration Number, if on another HMA protocol.Origin:COMS or PIBC#:
I. ADMINISTERED SUBSTANCES
1. Will you administer any substances to your animals during the course of the life of the animal(s)? Yes ☐ No ☐
If "NO", proceed to Section J
2. If “YES”, list all administered substances here and categorize (refer to http://toxnet.nlm.nih.gov/):
Do NOT include analgesia/anesthesia here, include as appropriate in Section B or Section J
Specify each substance:a. Chemical carcinogen:b. DEA Controlled substance:c. Toxic substance:d. Radioisotope: license #:
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e. Microbiological agent*:f. Human or animal cell lines** (including tumor cell lines):g. Human embryonic stem cells (hESC)***h. Diluents or carriers:i. Other (Including monoclonal antibodies and serum-derived products. Do not include
anesthestics or analgesics):
☐ I certify that BOTH animal-derived tumor or tissue cell lines and passaged human tumor or cell lines will be tested for biological contaminants prior to use, to prevent introduction of adventitious or zoonotic disease according to the following guideline:
HCCM must be contacted if you will be using substances potentially harmful to humans in the animal facilities, see the Mandatory Risk Disclosure page.
3. For substances potentially harmful to either animal subjects or staff:
a. The use of *microbiological agents must be approved by the Committee on Microbiological Safety (COMS) or Partners Institutional Biosafety Committee (PIBC). Contact a Biosafety Officer at EH&S at 617-432-1720 (HMS) or 800-825-5343 (BWH) for the necessary forms. BWH investigators must obtain approval from the Partners IBC; registrations can be completed online: https://insight.partners.org or contact the PIBC for assistance at [email protected]
Provide the COMS/PIBC Registration #
b. The use of **human or NHP tissues or cells must be approved by COMS or PIBC. Contact a Biosafety Officer at EH&S at 617-432-1720 (HMS) or 800-825-5343 (BWH) for the necessary forms. BWH/MGH investigators must obtain approval from the Partners IBC; registrations can be completed online: https://insight.partners.org or contact the PIBC for assistance at [email protected]
Provide the COMS/PIBC Registration #
c. Protocols involving the use of ***hESC must also be sent to the Harvard University Embryonic Stem Cell Research (ESCRO) Committee for review and approval (617-495-9829) and the Office of Research Compliance must be notified (617-432-3884).
BWH PI’s – Contact the Partner’s ESCRO Committee (617-424-4171).Children’s PI’s – Contact the Children’s ESCRO Committee
d. IACUC approval CANNOT be granted until ESCRO approval is in place. Have you submitted an ESCRO protocol application? Yes ☐ No ☐
List your ESCRO Protocol Number here: ___________________
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4. Specify the following for each substance listed above (copy this table and complete separately for each substance):
AGENT:
VERTEBRATE SPECIES to which agent will be administered:
a. Volume, Concentration, Route of administration:b. Anatomic Site of injection(s) (e.g. tail vein, thigh muscle) (if applicable):c. Number of Injection(s) (if applicable):d. Frequency of injection(s) (if applicable):e. Effects of the agent or substance on research animal:f. Length of survival time after initial administration:g. Will the animal experience pain, distress or illness as a result of the substance
administered? Yes ☐ No ☐
If "YES", describe the procedures to alleviate the pain, distress or illness:
h. Is this compound pharmaceutical grade (USP)? Yes ☐ No ☐ N/A ☐
If NO, the following standards should be followed for all non-pharmaceutical grade agents:
pH: Agents will be made pH appropriate for the route of administration. Purity: The highest available purity grade will be used. Stability: If stability information is unavailable, the agent will be prepared
immediately prior to use. Sterility: The agent and equipment will be sterilized (e.g. filtered, autoclaved,
etc.) prior to parenteral administration. Labeling/Storage: Agents will be labeled with the name, concentration, and
preparation date. Expiration: Agents will be used within the expiration date. Compounds will be
used within 30 days of preparation.
☐ I agree to abide by the above standards.☐ I will deviate from the above standards in the following way(s):
5. Specify the following for all HAZARDOUS/INFECTIOUS agents:
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You must complete the following section for all studies involving HAZARDOUS/INFECTIOUS agents:
Route of Administration or Delivery MethodShed in: Blood ☐ Saliva ☐ Urine ☐ Feces ☐
Location where agent will be administered:Building:Room:
Duration of Animal Survival Post treatment:
Animal Housing Location Post-treatment:
Special Housing Request:
Has contact with HCCM and EH&S been established for use of hazardous agents?Yes ☐ No ☐State the terms of, or describe the agreement with HCCM and EH&S on procedures and precautions for use of hazardous agents:
6. Pharmaceutical-grade compounds
a. Will you be using compounds, including diluents, that are not pharmaceutical-grade?Yes ☐ No ☐
In keeping with the USDA and OLAW guidance, it is the policy of the IACUC that pharmaceutical-grade compounds, including diluents and carriers, must be used in experiments instead of non-USP drugs or reagents, even in acute procedures. Non-pharmaceutical-grade chemical compounds, diluents and carriers may only be used after specific review and approval by the IACUC. The IACUC may consider scientific necessity or non-availability of an acceptable veterinary or human pharmaceutical-grade product as sufficient reasons for accepting the use of non-pharmaceutical grade drugs or diluents in animal experiments. Cost savings alone is not a sufficiently compelling reason to justify use of a potentially ineffective or toxic non-pharmaceutical-grade substance.
b. If you are administering ANY non-pharmaceutical grade substances, list each in the table below and check appropriate box a., b., or c. Microbiological agents need not be listed in this section.
Compound Identify need for non-USP substance
☐ a. Non-commercially available experimental compound
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☐ b. Only non-USP grade is commercially available and no other alternatives exist
☐ c. USP grade drug or alternative is available, but a non-USP grade is requested
c. If pharmaceutical-grade compounds are available and you are NOT administering pharmaceutical-grade compound(s) to your animal(s), (c is checked) you must also complete the detailed scientific justification column below.
The IACUC expects researchers to justify the use of a non-pharmaceutical grade drug, compound, diluent, or carrier and to demonstrate consideration (and rejection) of pharmaceutical grade alternatives for experimental reasons. Reasons related to the cost of USP grade products are generally deemed insufficient on those grounds alone to justify substitution of non-pharmaceutical grade alternatives for USP drugs and reagents. Provide sufficient detail and present the respective advantages and disadvantages of the pharmaceutical and non-pharmaceutical-grade compound/chemicals.
Describe how rejected alternatives will negatively impact outcomes or measurements. Describe how experimental logistics have influenced the choice of the chosen agent. Address any issues of animal or human safety, compound efficacy related to the experimental aim, and
address the possibility of inadvertent introduction of research-complicating variables.
Provide detailed scientific justification
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J. SURGERY Will you perform surgery? (See the HCCM website for specific requirements.) Yes ☐ No ☐
If “NO,” proceed to Section L
If “YES”, answer the following:
1. ANESTHESIA- complete table for each proposed surgery (for non-survival procedures, “duration” is the interval between induction and death).
Procedure Name Species Anesthetic(s) Dose(s) & Route Duration (Minutes/Hours)
2. NON-SURVIVAL SURGERY- animals that will not recover from surgery (DO NOT include description of “euthanasia only” animals):
a. Specify non-survival surgery (e.g. perfusion, laparotomy & organ harvest, terminal embryo harvest):
b. Location (building & room):
c. Describe surgical site prep:
d. Describe non-survival surgery in detail (exclude anesthesia methods):
e. Euthanasia method for non-survival surgery:
3. SURVIVAL SURGERY - animals that will recover from surgery
a. Specify MAJOR SURGERY (entering body cavity, e.g. laparotomy, craniotomy or creation of permanent physiological defect):
b. Specify MINOR SURGERY (cut-downs, needle aspirations, etc.):
c. Identify the location of surgery. Specify building and room:
d. Identify the location of housing after surgery. Specify building and room:
e. Describe surgical site prep:
f. Describe the surgical procedure in detail (include anatomic site, incision/manipulation, and closure details, exclude anesthesia methods):
g. Explain why it is necessary for the animals to recover from surgery:
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h. State the anticipated survival interval following surgery that is required to complete experimental objectives:
i. Describe your plan for intra-operative AND post-operative monitoring of the animals for the first 4 post-operative days. Laboratory personnel are responsible for post-operative monitoring.
j. Describe any impairment that may result after the surgery. List potential post-operative complications or sequelae. Describe your plans to handle them:
k. Describe how you will treat post-operative pain, and state analgesic, dose and route: If multiple survival procedures are described above, specify the analgesic therapy for each.
Analgesic:Dose:Route:Interval between administration of drugs:Duration of analgesic therapy (Post-operative):
**You must administer analgesia for 48 hours after major procedures and for 24 hours after minor procedures (the dosing regimen will depend on the analgesic
agent). Reproductive surgery is considered a major surgical procedure. **
K. MULTIPLE SURVIVAL SURGERY
1. Will any animals undergo more than one survival surgical procedure? Yes ☐ No ☐ See the Guide, page 30:
(NOTE: if the second surgery is a non-survival surgery, answer ‘NO’ and proceed to Section L)
2. If “YES”, complete the following
a. List procedures in chronological order:
Surgical procedure Major Minor Interval before subsequent procedure
1.
2.
3.
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b. Explain why it is scientifically necessary to have the animal(s) undergo more than one SURVIVAL surgical procedure:
L. PAINFUL PROCEDURES
1. Will your study be concerned with the effects of TRAUMA? Yes ☐ No ☐
2. Will your study be concerned with the effects of BURNS? Yes ☐ No ☐
3. Will you be studying PAIN? Yes ☐ No ☐
4. Will you be using ELECTRIC SHOCK? Yes ☐ No ☐
If “YES” to any of the above, complete the following:
a. Describe the procedure in detail:
b. i. State your proposed procedures to avoid or alleviate the pain:
ii. State the anesthetic or analgesic agents, dosages, route and intervals between doses to be used for each species:
iii. Provide justification if you do not intend to use anesthetic or analgesic:
c. Describe the monitoring schedule:
d. State the impairment that can be expected from the procedure:
e. State the duration of survival after the procedure:
5. Will you be using paralytic agents* or performing paralytic procedures? Yes ☐ No ☐
(*Examples include, but are not limited to: Succinylcholine chloride, Decamethonium bromide, D-Tubocurarine chloride, Gallamine triethiodide, Pancuronium bromide, Vecuronium bromide, Atracurium besylate, Mivacurium, Doxacurium, and Pipecuronium.)
If "YES", complete the following:
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a. Describe the procedure/use of agent in detail:
b. Provide scientific justification for the use of paralytic agent/procedure(s):
c. Describe the impairment expected from the procedure:
d.i. Describe your proposed procedures to avoid or alleviate pain:
ii. State the criteria to evaluate pain (inability to reach food and water, etc) and what will be done to alleviate pain:
iii. State the anesthetic or analgesic agents and dosages to be used for each species:
e. State whether the animals will be able to urinate/defecate properly, and if not, what will be done to assist the animal:
f. Describe your monitoring plan.
g. State the duration of survival after the procedure:
M. BEHAVIORAL CONTROLS, RESTRAINT OR ANALYSIS OF BEHAVIOR ON ALERT ANIMALS
1. Will your study require a physical restraint device of any type? Yes ☐ No ☐
Physical restraint includes, but is not limited to: chairs, pie chambers, tail first restrainers, rabbit restrainers, tethers, jackets, etc. See the Guide , page 29. If "YES", complete the following:
a. Describe restraint device:
b. Anticipated duration of restraint:
c. Explain choice of restraint method, include considered alternatives:
d. Describe acclimation to restraint device prior to experiment (if not acclimating, please justify):
e. What will happen if any animals fail to acclimate to the restraint?
f. Describe observation/monitoring of animals while restrained (include any additional means used to ensure the welfare of the animal while in the restraint, include measures to be taken if animals appear to be in distress):
g. Prolonged restraint (Prolonged restraint is defined as any restraint other than brief restraint for the purpose of performing routine clinical or experimental procedures.) Provide justification for why
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prolonged restraint is necessary for your studies (include search for alternatives in Section P):
2. Will your study involve experiments on alert animals, behavioral control, or analysis of behavior? Yes ☐ No ☐
If "YES", complete the following:
a. If behavioral control, with use of food or water deprivation, state the duration of food or water deprivation and means to ensure the welfare of the animal:
b. If behavioral control with psychotropic drugs and withdrawal, specify duration, monitoring plan during withdrawal and means to ensure the welfare of the animal:
N. DIET AND OTHER ENVIRONMENTAL CHANGES
1. Will this study involve any of the following environmental changes or diet changes?(NOT including deprivation to prepare for surgery)?
YES NOa. Food deprivation? ☐ ☐b. Water deprivation? ☐ ☐c. Temperature changes? ☐ ☐d. Changes in the light cycle? ☐ ☐e. Other diets or changes (specify): ☐ ☐
2. If “YES” to any of the above, complete the following:
a. Brief description of the experiment:
b. Effect on the animal’s health - be specific:
c. Duration of the experiment:
d. Specify duration of water or food deprivation and provide justification:
e. Describe the specific observation schedule:
f. Specify criteria for removal from the experiment:
Comply with HCCM policy, see: https://hccm.med.harvard.edu/restriction-of-food-or-fluid-for-laboratory-animals
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O. DRUG AND SURGICAL SUPPLY MANAGEMENT PLAN
It is federal policy that expired drugs cannot be used in animal research. However, expired surgical supplies (e.g. sutures, catheters) may be used for non-survival procedures only. All laboratories or departments must create a “medical drugs and surgical supplies management plan” describing the laboratory’s mechanism to monitor and dispose of or properly mark all expired drugs and surgical supplies on a regular basis. Expired surgical supplies may be labeled for ‘non-survival’ use.
Laboratories which use drugs or surgical supplies must make their management plan available in the laboratory for any regulatory inspections such as the semi-annual IACUC inspection, or any USDA or AAALAC inspections. The following are suggested methods for managing your medical drugs and surgical supplies:
Mark a computer calendar program (e.g. Outlook) with expiration date check reminder Color or sticker-code materials Conduct appropriate short-term ordering and stock rotation Appoint an expiration date monitor within the lab Train all research staff in medical drugs and surgical supply storage Consolidate all medical supplies into a single area Conduct monthly laboratory checks Any other method appropriate for your laboratory/department
a. Check here to indicate that you will implement a drug & surgical supplies management plan: ☐
b. State the building and room number where drugs will be stored:
BWH Investigators must contact the BWH Office of Research Compliance for more information and guidance (617-732-5761).
P. ALTERNATIVES TO PAIN AND DISTRESS
NOTE*: Current legislation in the United States decrees that all proposed use of laboratory animals should be subject to review to determine whether such use appears to be scientifically and ethically justifiable. Investigators seeking to use animals in research thus have a legal and moral obligation to replace, reduce and refine laboratory animal experimentation wherever possible. The Animal Welfare Act requires that all procedures involving animals be reviewed by an institutional animal care and use committee (IACUC). The IACUCs are required to ensure that the protocols are worthwhile, that they
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use the minimum number of animals necessary, and that the investigators document that they have adequately considered alternatives to any procedure that causes more than momentary pain or distress (either with or without the use of anesthetics). It is the responsibility of the investigator to try to find ways whenever possible to employ research strategies and experimental designs that:
REDUCE Find ways to obtain comparable levels of information from the use of fewer animals in scientific procedures or obtain more information from a given number of animals so that, in the long run, fewer animals are needed to complete a given research project or test. The number of animals used should be the minimum that is consistent with the aims of the experiment. Examples for reducing the number of painful procedures include decreasing the number of time points for blood sampling or using saliva sampling rather than tail biopsy.
REFINE Employ methods that alleviate or minimize potential pain and distress and enhance animal well-being. Much potential pain and distress can be avoided or alleviated with the proper use of anesthetics, analgesics, and tranquilizers. Examples of refinement include the use of current anesthetics or analgesics to improve pain management or using new surgical materials, sutures or equipment
REPLACE Consider methods that permit a given purpose to be achieved without conducting experiments or other scientific procedures on animal. Such alternative methods and approaches include improved storage, exchange, and use of information about previous animal experiments to avoid unnecessary repetition of animal procedures; use of physical and chemical techniques and predictions based upon the physical and chemical properties of molecules; use of mathematical and computer models; use of organisms with limited sentience such as invertebrates, plants and microorganisms; use of in vitro methods including sub-cellular fractions, tissue slices, cell suspensions, and perfused organs. In vitro methods are increasingly used as the methods of choice in place of animal studies because they offer the best scientific approach. Such replacement alternatives include using in vitro methods of antibody production rather than the ascites model.
1. Will any of the proposed procedures cause more than slight or momentary pain or distress to the animals?(**IMPORTANT: If your procedures are listed in USDA Category D or E (from Section A - Checklist) and/or include anything other than observation, breeding, venipuncture, simple injection, or euthanasia followed by tissue/organ harvest, then you MUST answer "YES" in accordance with USDA regulations for covered species and PHS and AAALAC policy for all species. This includes tail biopsy for genotyping). Yes ☐ No ☐
If "YES", then you must explain below by answering P.2 and/or P.3 why you cannot use alternative procedures that might cause less pain or distress. If you are performing tail biopsy for genotyping, you must complete section P3 to document why/if alternatives (stool, pin prick and blood sample collection on filter paper, saliva samples) are not feasible. Also you should confirm that you will follow the tail biopsy SOP, available at: http://collaborate.hms.harvard.edu/display/IACUC/317.01.
The USDA requires that you make a good faith effort to try to find alternatives to or look for refinements for each painful or distressful procedure.
2. Alternatives do NOT exist. You know this because the following sources were consulted:
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a. Literature search: Complete a search for each potentially painful and/or distressful procedure used in this amendment in the table below (copy & paste for each additional painful procedure).
NOTE: Key words should specify the painful or distressing procedures, not the model being studied. You must include Pubmed and at least one of these free alternative resources websites in your search:
ALTWEB (Alternatives to Animal Testing on the Web) http://altweb.jhsph.edu
NAL Animal Use Alternatives Thesaurus http://agclass.nal.usda.gov/agt.shtml
Animal Welfare Institute http://www.awionline.org/lab_animals/index.htm
Procedure:
Database(s) Searched:
Date searched:
Keywords Used:
Years Covered by the Search:
Include one or more of the following (optional):
b. Meetings or conferences (names and dates of meetings attended):
c. Periodicals (names of periodicals or journals read on a regular basis):
d. Consultation with colleagues (names and credentials of colleagues, such as MD, PhD, dates of consultations and nature of discussions):
AND/OR
3. Alternatives do exist [or one is found through the search], but:
a. Explain why alternative experiments are unsatisfactory:
b. Explain the steps you will take to assure that the animals will not be subject to any more discomfort
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than is absolutely necessary:
c. Describe the methods used to ensure that the duration of the discomfort will be as short as possible:
d. Explain your plans to monitor and correct problems (e.g., by euthanasia):
Q. MINIMIZATION OF DISCOMFORT, PAIN AND DISTRESS
Summarize your procedures as follows to avoid or minimize any discomfort, pain or distress the animals may experience.
1. Describe observation schedule:
2. Provide criteria for determination of pain and distress:
3. Describe what actions will be taken to relieve pain and distress:
4. Provide the experimental endpoint(s) at which euthanasia will take place:
5. Describe humane endpoints that will supercede experimental endpoints for clinical conditions or unexpected events. (Veterinary guidance available at http://collaborate.hms.harvard.edu/display/IACUC/317.01. Must be logged in to view.)
THE USDA REQUIRES THAT YOU LIST THE NAMES AND DOSAGES OF ANALGESICS OR TRANQUILIZING DRUGS TO BE USED FOR EACH SPECIES OF ANIMAL UNDER STUDY.
[Note: If no analgesia is given post-operatively for major surgery, then you will need to check ‘Category E’ as appropriate in the Section A Checklist
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R. DUPLICATION OF RESULTSIn accordance with the USDA Regulations (9 CFR Part 1-3) and the Animal Welfare Act, the HMA
Standing Committee on Animals is required by Federal Law to obtain the following assurance from you:
1. Do the activities described in this protocol application form UNNECESSARILY duplicate previous experiments whether your own or another investigators’ experiments?
Yes ☐ No ☐
2. If “NO”, indicate below how you determined that these activities do not unnecessarily duplicate previous experiments (literature searches, attendance at national or international meetings, etc.).
a. Meetings or conferences. List names and dates of meetings attended:
b. Periodicals. Names of periodicals read on a regular basis:
c. Consultation with colleagues. List names and credentials of colleagues (i.e., MD, PhD), dates of consultations and nature of discussions:
d. Literature searches. Names of literature searches completed and key words:
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S. ASSURANCE/SIGNATURE PAGES
1. ASSURANCE: As Principal Investigator, I certify the following:
I assume responsibility for the work described here in this proposal.
I understand that I am responsible for ensuring that I and my staff will comply with the policies governing the care and use of animals at Harvard Medical School. It is understood that these policies include applicable local, State, and Federal policies and regulations.
If this is for the three-year review of a previously approved protocol, I confirm that this application includes all relevant animal procedures, and I understand that any procedures or other changes in amendments not included in this application will no longer be approved, and that any previously approved Exception to Standards will be resubmitted for IACUC review.
I confirm that the information described in this proposal is correct and accurately reflects the information in the specific grant application(s) or fellowships cited on the Face Page of this protocol form.
I understand that I am responsible for ensuring that I and my staff are familiar with proper handling, experimental and restraint techniques required for the species used or will seek advice and assistance from the staff of the Harvard Center for Comparative Medicine. In addition, I confirm that all personnel listed in section C of this application shall review the protocol following IACUC approval and prior to participation in any of the activities described therein.
I agree to abide by the requirements for the use of pharmaceutical-grade compounds (Section I.6.).
If I perform survival surgery, I agree to abide by the following requirements for sterile technique (this does not apply to frogs or fish):
I agree to prep animals for surgery by clipping hair and disinfecting the skin I agree to maintain sterile conditions and a sterile field at all times I agree to use sterile instruments at all times
This research does not represent unnecessary duplication of previous studies.
I understand that HMA IACUC approval is valid for one year only; that approval must be renewed annually; that every third year the IACUC must perform a new review of my protocol before work may proceed.
Finally, I realize that failure to adhere to HMA IACUC policies relating to animal care and use may result in suspension or revocation of permission to perform animal research in Harvard Medical School facilities.
2. SIGNATURES:
Principal Investigator’s signature: Date:
Sponsor’s signature: Date: (Sponsor required if PI is not of ‘Faculty’ rank)
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Department Head’s signature: Date: (Required for all submissions)
Department Head print name: Date:
The following statement must be included within any new or competing grant application:
The Harvard Medical School animal management program is accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care, International (AAALAC), and meets National Institutes of Health standards as set forth in the 8th edition of the Guide for the Care and Use of Laboratory Animals (National Research Council. "2 Animal Care and Use Program." Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press, 2011). The institution also accepts as mandatory the PHS Policy on Humane Care and Use of Laboratory Animals by Awardee Institutions and NIH Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training. There is on file with the Office of Laboratory Animal Welfare (OLAW) an approved Assurance of Compliance (A3431-01).
