Harvard Medical Area (HMA) Standing Committee on Animals – “Established in 1907”

Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 Harvard Medical Area (HMA) Standing Committee on Animals – “Established in 1907” 180 Longwood Avenue, Suite 113 Boston, MA 02115 Tel: 617-432-3192 Fax: 617-432-3169 http://www.hms.harvard.edu/orsp/ iacuc@hms.harvard.edu 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 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: Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 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 Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 MRD: MANDATORY RISK DISCLOSURE - Hazardous substance disclosure sheet STOP YOU MUST RECORD AND CHECK OFF THE FOLLOWING PERTAINING TO YOUR SUBMITTED PROTOCOL: FAILURE TO DO SO WILL RESULT IN A DELAY IN PROTOCOL APPROVAL Note- items checked below must correspond with the information provided in your protocol **MUST CHECK (X) ALL THAT APPLY** Check: Procedures: Chemical carcinogen(s) Identify specific agent(s): Route(s): Toxic substance(s) Radioisotope license #: 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 ABOVE This 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 Note: YOU MUST NOTIFY THE HCCM WHEN YOU BEGIN WORK WITH THESE AGENTS Contact the HCCM Coordinator of Special Projects via phone at: 617-432-4658 or mailto:hccm@hms.harvard.edu PI must print and sign below (mandatory; a proxy will not be accepted): P.I Name _ P.I Signature Date / / Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 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  The Deadline for submission of a New AEP, 3-Year Renewal, or Amendment is 5:00 p.m on or before the 1st business day of that month Failure to comply will result in the delay of review for an additional month You must submit an electronic (MSWord) version of your protocol  Fax Signature pages to 617-432-3169  Submit 3-Year Renewal two months prior to the protocol’s expiration date to avoid a lapse in research Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 A ANIMAL USAGE 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 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.) 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 P CATEGORY 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: ascites 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 Footpad injections (antibody production or microorganism) Introduction of illness (including the administration of toxins, microbiological agents, clinical transgenic phenotypes) Lavage Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 Multiple MAJOR survival surgery Multiple MINOR survival surgery Non-survival surgery Physical trauma Survival surgery: MAJOR Survival surgery: MINOR Tail biopsy for genotyping Toxic substances (e.g streptozotocin, MPTP, cisplatin, doxycycline) Transgenic mouse production (embryo manipulations to produce novel strains, etc Tumor induction or implantation Unusual or prolonged restraint (e.g pie chamber, chair) Use of paralytic agents 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 Death-as-an-endpoint** studies Lethal dose studies** Noxious stimuli from which there is no escape Pain study Withholding of post-operative analgesia for any surgical procedure Total number of animals to be used in Category E **You must follow the Guidelines for ‘Death as an Endpoint’ or ‘Lethal Dose’ studies at: https://www.hms.harvard.edu/ORSP/Secure/animal_guidelines.html 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 and/or separate exemption form Section D.4 Single housing of animals Section D.4.c Withholding of enrichment from singly housed animals Section E & Exemption Euthanasia methods that are not consistent with the AVMA Guidelines Section G & Exemption Ascites collection Section I.7 Non-pharmaceutical grade drugs and other compound suspensions Section J.2.k & Exemption Withholding postoperative analgesia Section N Food/water withholding or scheduling (not ad-lib) Exemption form Smaller caging than the recommended space allotment (e.g.: CLAMS) Exemption form Deviations from housing density standards Exemption form Toe-clipping for identification purposes Section N Deviations from the 12/12 hour light/dark cycle Exemption form Any other exceptions to the standards of the PHS Policy, the Guide, or the AWA - Please specify: Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 JUSTIFICATION FOR THE USE OF ANIMALS Federal 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: 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 Procurement, Allocation, and Reuse Committee 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 Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013  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 not guarantee Committee approval 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 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 only in Section J FOR 3-YEAR REVIEWS: a Progress Report: Briefly describe your RESULTS from the past years and explain how the present application continues this work: b For USDA-covered species (not mice, rats, or birds), describe any unexpected animal deaths in the course of your experiments: c List publications and meetings in the past three years: d Changes (briefly summarize any CHANGES in the protocol in the past three years, i.e species, numbers, procedures, etc.): Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 FOR ALL (NEW and 3-YEAR Review) protocols FLOWCHART - you must diagram your procedures in a flowchart  Present 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  Include animal numbers  For guidelines on creating your flowchart click here  USDA covered species users note: This flowchart will be included with the medical record of each animal Present your flowchart here: Harvard Medical Area Page 10 of 35 Protocol # Revised 2/1/2013 C PERSONNEL Will the principal investigator perform the procedure(s)? Yes No Personnel List:  Provide the name(s), title(s) and qualifications for each person, including the PI, who will work with the animals  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 the training  NOTE - ALL current personnel must sign the last page of this form, Section S to gain access to the facilities NEPRC clinical staff members that not have substantial, direct, intellectual contributions to the work not need to be listed or sign this form unless they are the PI but must sign a confirmation compliance statement annually  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 Name Title/Job Classification 10 Personnel Training: Qualification/ Experience Phone Email Address Harvard Medical Area Page 22 of 35 Protocol # Revised 2/1/2013 Pharmaceutical-grade compounds a Will you be using agents 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 nonpharmaceutical-grade compound b If you are administering ANY non-pharmaceutical grade drugs, diluents or carriers, list each in the table below and check appropriate box a., b., or c Agent X Identify need for non-USP compound a Non-commercially available experimental compound 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 pharmaceuticalgrade 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 Provides detailed scientific justification Harvard Medical Area Page 23 of 35 Protocol # Revised 2/1/2013 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: Non-survival SURGERY- animals that will not recover from surgery (DO NOT include description of “euthanasia only” animals): Species Anesthetic Dose Route a Non-survival surgery details (describe non-survival surgery in detail): b Euthanasia for non-survival surgery (specify euthanasia method and procedure for these animals): c Location of non-survival surgery Building: Room: 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 the surgical procedure in detail: f Identify anesthetic agents and dosages in mg/kg (body weight), routes of administration for each species: g Explain why it is necessary for the animals to recover from surgery: h Describe your plan for post-operative monitoring of the animals for the first post-operative days State who will be responsible for the monitoring: i Describe any impairment that may result after the surgery List potential post-operative Harvard Medical Area Page 24 of 35 Protocol # Revised 2/1/2013 complications or sequelae Describe your plans to handle them: j State the anticipated survival interval following surgery that is required to complete experimental objectives: k Describe how you will treat post-operative pain, and state analgesic, dose and route: 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 Will any animals undergo more than one survival surgical procedure? See the Guide, page 30: Yes No (NOTE: if the second surgery is a non-survival surgery, answer ‘NO’ and proceed to Section L) If “YES”, complete the following a List procedures in chronological order: Surgical procedure Major Minor Interval before subsequent procedure b Explain why it is scientifically necessary to have the animal(s) undergo more than one SURVIVAL surgical procedure: Harvard Medical Area Page 25 of 35 Protocol # Revised 2/1/2013 L PAINFUL PROCEDURES Will your study be concerned with the effects of TRAUMA? Yes No Will your study be concerned with the effects of BURNS? Yes No Will you be studying PAIN? Yes No 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 not intend to use anesthetic or analgesic: c State who will monitor the animals after the procedure and how often: d State the impairment that can be expected from the procedure: e State the duration of survival after the procedure: 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: 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: Harvard Medical Area Page 26 of 35 Protocol # Revised 2/1/2013 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 Identify who will monitor the animals after the procedure/agent administration and how often: g State the duration of survival after the procedure: M BEHAVIORAL CONTROLS, RESTRAINT OR ANALYSIS OF BEHAVIOR ON ALERT ANIMALS 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 (Brief restraint for the purpose of performing routine clinical or experimental procedures need not be described here ) Provide justification for why prolonged restraint is necessary for your studies (include search for alternatives in Section P): Will your study involve experiments on alert animals, behavioral control, or analysis of behavior? Yes No If "YES", complete the following: Harvard Medical Area Page 27 of 35 Protocol # Revised 2/1/2013 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 Will this study involve any of the following environmental changes or diet changes? (NOT including deprivation to prepare for surgery)? YES a b c d e NO Food deprivation? Water deprivation? Temperature changes? Changes in the light cycle? Other diets or changes (specify): 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 and identify the personnel who will monitor the animals: f Specify your monitoring criteria: Comply with HCCM policy, see: https://ARCM.med.harvard.edu/secure/documents/pdfs/policy_food_or_fluid_restriction.pdf 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 Harvard Medical Area Page 28 of 35 Protocol # Revised 2/1/2013 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 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: Harvard Medical Area Page 29 of 35 Protocol # Revised 2/1/2013 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 wellbeing 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 * excerpted from Environmental Health Perspectives, August 1996 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: https://arcm.med.harvard.edu/secure/documents/pdfs/sop_317.01_partial_tip_excision_biopsy_mice.pdf 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 Alternatives NOT exist You know this because the following sources were consulted: Harvard Medical Area Page 30 of 35 Protocol # Revised 2/1/2013 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/altrntvs/index.htm 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: 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 Alternatives 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 than is absolutely necessary: Harvard Medical Area Page 31 of 35 Protocol # Revised 2/1/2013 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 Describe observation schedule: Provide criteria for determination of pain and distress: Describe what actions will be taken to relieve pain and distress: Provide the experimental endpoint(s) at which euthanasia will take place: Describe humane endpoints for clinical conditions or unexpected events (Veterinary guidance available at https://arcm.med.harvard.edu/secure/documents/pdfs/humane_endpoints.pdf): 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 Species Analgesic Dose Route R DUPLICATION OF RESULTS In 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: Harvard Medical Area Page 32 of 35 Protocol # Revised 2/1/2013 Do the activities described in this protocol application form UNNECESSARILY duplicate previous experiments whether your own or another investigators’ experiments? Yes No If “NO”, indicate below how you determined that these activities 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: Harvard Medical Area Page 33 of 35 Protocol # Revised 2/1/2013 S ASSURANCE/SIGNATURE PAGES The investigator and research staff assigned to this protocol are required to sign this form, thus certifying that they: a have read the protocol, b are familiar with all of its requirements, and the requirements of all rules regulations and policies governing the care and use of animals relating to their activities; c have had the opportunity to ask questions about it; d understand that all animal experimentation not described in the protocol is prohibited in the absence of an approved amendment; e are authorized to conduct procedures involving animals under this proposal; f have completed the institutionally required investigator training, and received training in: the handling, and care of this species; aseptic surgical methods and techniques (if necessary); the concept, availability, and use of research or testing methods that limit the use of animals or minimize distress; the proper use of anesthetics, analgesics, and tranquilizers (if necessary); and procedures for reporting animal welfare concerns as required by Section 2.32 of the Animal Welfare Act NB: Veterinary and veterinary tech staff members that not have substantial, direct, intellectual contributions to the work not need to sign below Name Signature Harvard Medical Area Page 34 of 35 Protocol # Revised 2/1/2013 CERTIFICATIONS: 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 I agree to abide by the requirements for the use of pharmaceutical-grade compounds (Section I.6.) If I am doing 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 SIGNATURES: Principal Investigator’s signature: Date: Sponsor’s signature: Date: (Sponsor required if PI is not of ‘Faculty’ rank) 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: Harvard Medical Area Page 35 of 35 Protocol # Revised 2/1/2013 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) ... Date / / Harvard Medical Area Page of 35 Protocol # Revised 2/1/2013 Introduction The Harvard Medical School Standing Committee on Animals was established in 1907 almost 60 years... 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... indicated section of the protocol and/or separate exemption form Section D.4 Single housing of animals Section D.4.c Withholding of enrichment from singly housed animals Section E & Exemption Euthanasia