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Introduction to the Use of Animals and the DLAM Facilities

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					Mandatory Animal Care and Use (MACU)
              Training
     Introduction to the Use of Animals
          and the DLAM Facilities




    Institutional Animal Care and Use Committee

      Tufts University & Tufts Medical Center
         Outline of Presentation

   Introduction to Federal and State Regulations
   Institutional Animal Care and Use Committee
    (IACUC)
   Division of Laboratory Animal Medicine
   Institutional Procedures
    Regulations Governing the Use of
          Animals in Research
   The Animal Welfare Act, passed in 1966, is overseen by
    the United States Department of Agriculture (USDA)
    http://www.aphis.usda.gov/ac/awa.html
   Public Health Service Policy was passed in 1985 and is
    overseen by the Office of Laboratory Animal Welfare at
    the National Institute of Health (OLAW at NIH)
    http://grants.nih.gov/grants/olaw/references/phspol.htm
   Massachusetts Research Animal Regulations (MDPH)
    was passed in 1985 and is overseen by the MA
    Department of Public Health
           The Animal Welfare Act
   Rats (genus Rattus), mice (genus Mus), birds, frogs, and fish
    are among those currently exempted from definition of
    “animal.”
   Horses and farm animals used in biomedical research and
    teaching are included in the definition of “animal,” but
    animals used for food and fiber only are exempt.
   Requires the Institutional Official (IO) of the Institution to
    appoint an Institutional Animal Care and Use Committee
    (IACUC)
   Compliance is ensured by unannounced annual inspections
    (at a minimum) of each registered Institution.
   Non-Compliance is managed by monetary fines and legal
    action against the Institution
   Institutions are required to submit an Annual Report to USDA
    with the species, numbers, and pain/distress categories used
    within that year.
    Public Health Service Policy (PHS)
 Defines “animal” as any live vertebrate
 Requires IO of Institution to appoint Institutional
  Animal Care and Use Committee (IACUC).
 Requires Institution to provide a written Assurance to
  the Office of Laboratory Animal Welfare (OLAW) at the
  National Institutes of Health (NIH) in order to receive
  funding from NIH.
 Each Institution has it’s own Assurance number on file
  with NIH. You must provide your campus’ Assurance
  number on your funding application.
 Requires that Institutions follow both of these
  publications to receive funding.
    ◦ PHS Policy on Humane Care and Use of Laboratory Animals
    ◦ Guide for the Care and Use of Laboratory Animals
      PHS Policy and OLAW
 Ensures compliance by requiring that each
  Institution’s Assurance is resubmitted and
  renewed every 4 years. Additional
  requirements for the Institution can be made
  by OLAW prior to renewal.
 Requires any “significant” issue of
  non-compliance to be reported to OLAW and
  to the funding component.
 For a definition of what OLAW considers
  significant non-compliance, go to:
  http://grants.nih.gov/grants/guide/notice-
  files/NOT-OD-05-034.html
Massachusetts Research Animal Regulations
 Massachusetts Research Animal Regulations were
  passed in 1985.
 Overseen by the MA Department of Public Health
  (MDPH)
 Requires each Institution to hold a license for research
  animal use
 The MA Commission of Public Health, or an agent
  designee, may visit and inspect the animal research
  institution that houses dogs or cats. These inspections
  may be unannounced and a facility could be inspected
  four times a year.
                       AAALAC
Association for the Assessment and Accreditation of
           Laboratory Animal Care, Int’l
 Accreditation is voluntary and is recognized as the highest
  standard of laboratory animal care
 Triennial visits to ensure that the Institution meets
  regulatory compliance and ensures high standards of
  laboratory animal medicine.
 Tufts Boston and Grafton are both AAALAC accredited.
  Each campus maintains its own accreditation number.
Regulatory oversight of animal
          research
               3 Rs of Russell and Burch
       The Principles of Humane Experimental Technique

   The concept of the “3Rs” are cited in the Regulations and are used in our
   Institutional policies as ethical guidance for the use of animals in research.
   Each is defined below:
Replacement -methods that avoid using animals, such as replacing animals with
  inanimate systems or replacing animals such as vertebrates with animals lower on
  the phylogenetic scale.
Refinement-modifications of husbandry or experimental procedures to enhance animal
  well-being and minimize or eliminate pain and distress.
Reduction-strategies for obtaining comparable levels of information from the use of
  fewer animals or for maximizing the information obtained from a given number of
  animals (without increasing pain/distress) so that fewer animals are needed to
  acquire the same scientific information.
  Investigators are discouraged from advocating animal re-use as a reduction strategy.
  Reduction should not be a rationale for re-using an animal that has already
  undergone experimental procedures, especially if the well-being of the animal
  would be compromised.
    Animal Welfare Concerns
 Whistleblower Policy-animal welfare concerns are
  handled with as much confidentiality as possible.
  Reporters are protected by law. (See Policy for detailed
  information and contact numbers.)
 Reports can be made to:
  ◦ Any member of the IACUC staff or committee
  ◦ The IACUC Chair
  ◦ Your Principal Investigator
  ◦ The Tufts anonymous hotline at 617-636-2492
    Institutional Animal Care and Use
           Committee (IACUC)
   Committee members are appointed by the
    Institutional Official, Peggy Newell

• IACUC Staff
    - Dr. Barry Goldin, Boston/Grafton IACUC Chair
    - Dr. L. Michael Romero, Medford IACUC Chair
    - Valerie Parkison, IACUC/IBC Manager
    - Stephanie Heller, IACUC Specialist
    - Ann Holm, IACUC Coordinator, Grafton campus
    - Terri Naumes, Regulatory Admin. Assistant
         Institutional Animal Care and Use
                Committee (IACUC)
The role of the IACUC is to oversee and routinely evaluate the Animal Care
  and Use Program (the Program), which includes:
   Ensure that the Institution is in compliance with all federal and state laws.
   Protect Institution’s Assurance, licenses, and accreditation so that grant
    funding can be received.
   Review and approve proposed animal use (via protocols) and of proposed
    changes (via amendments) to animal use.
   Regular inspections of centralized facilities and laboratories approved for
    animal use.
   Regular review of program requirements, such as training, occupational
    health and safety, grant verifications, etc.
   Establishment of a mechanism for receipt and review of concerns involving
    the care and use of animals at the institution. The IACUC is authorized to
    suspend any animal activity in order to protect the animals’ welfare.
   Submit reports of program evaluation to government agencies and to the
    Institutional Official (IO).
                 IACUC Constitution
Tufts Institutional Care and Use Committee membership includes:

 Practicing scientists experienced in research involving
  animals. Each scientist represents a different
  department/division.
 Attending veterinarian(s).
 One member from a nonscientific background from Tufts
  University.
 One public member to represent general community
  interests in the proper care and use of animals.
            The IACUC and its Institution
   The IACUC serves as the advisory Committee on all matters
    concerning the humane and ethical care of research animals.
   The IACUC investigates and ensures resolution to compliance
    issues involving the care and use of research and teaching
    animals.
   The IO is the institutional representative of the IACUC and is
    asked to assist when:
     1) the IACUC votes an issue of non-compliance as
         “significant” and therefore reportable to federal agencies
     2) federal reports are submitted
     3) financial resources are necessary to maintain
        compliance in the program or the facilities.
Information for the IACUC Protocol
 Project Objective and Justification
   Required to describe proposed research in lay terminology to represent the public’s
    interest in publicly-funded work.
   Should include hypothesis, explicit goals, potential contributions to human and/or
    animal health & the advancement of biomedical knowledge.
   Justify use of animals & why non-animal alternatives cannot be utilized.
   Justify choice of species & describe unique characteristics that each species, stock, or
    strain has that are necessary for your objectives.
   Justify how the number of animals requested was determined. Including a brief
    description of statistical analysis, including tests, power and probability levels utilized, if
    applicable.
 Experimental Plan and Design
   Describe all procedures and time intervals between procedures. Include specific
    information on how study duration and endpoints will be determined. Indicate how all
    procedures and compounds administered will affect animal health & ultimate disposition
    of animals.
   Requires approval of all procedures performed on animals. This includes procedures that
    are not “experimental”, such as tail biopsies, post-procedural care, etc.
   Category Definitions – you must place your animals into one of the following three
    categories and justify the choice of category:
     USDA Category C: procedures with minimal, momentary, or no distress
     USDA Category D: use of appropriate anesthetics, tranquilizers, or analgesics to alleviate
     pain and/or distress
     USDA Category E: animals may experience unrelieved pain and/or distress without
     intervention
            IACUC Protocol Form
 Search for Alternatives
   Federal regulations mandate you describe how the lack of alternatives were
    verified for each potentially painful/distressing procedure (Cat. D or E). Must be
    in the form of a narrative description including the date search was conducted,
    database searched, years covered by search, keywords or search strategy, and
    number of references found.
Personnel Qualifications
   Federal law requires IACUC evaluation of training and qualifications of
    personnel intending to use live animals in research or teaching. Personnel must
    understand the basic needs of species utilized, use proper techniques when
    handling each species, select methods that minimize distress, provide proper pre-
    and post-procedural care to animals, use aseptic surgical techniques, & use
    anesthetics/tranquilizers appropriate for each species.
 Authorization of the Principal Investigator
   PI must confirm that information is accurate and conforms with institutional and
    regulatory policies on using animals in research & teaching, that alternatives have
    been considered, that project doesn’t unnecessarily duplicate previous
    experiments, that personnel have agreed to participate & are aware of approved
    procedures, & are instructed in humane care, handling, and use of animals. They
    must confirm that no changes will be made to procedures, care, or housing with
    prior IACUC approval, & that failure to comply w/ IACUC policies and
    procedures will jeopardize Tufts Animal Welfare Assurance on file with the NIH,
    and may lead to revocation of privileges to conduct animal research.
 Euthanasia (or Final Disposition)
   Must justify how euthanasia is performed and provide details. A secondary,
    physical method of euthanasia is required if an inhalant is used (CO2, isoflurane,
    etc).
             Protocol Amendments
   All changes in protocol must be approved by the IACUC
    prior to implementation, which includes:
    - change in procedure
    - change in species
    - change in animal number

   The amendment form (Supplement A) is used to make
    these changes.

   Amendments are reviewed by designated reviewers and
    generally require one week to process.
    - Some amendments are considered significant changes to the original
      protocol and require Full Committee review rather than Designated
      Review. These include; change in species, a significant increase in
      animal numbers, additional Category E animals, or addition of a
      Category E procedure.
         Use of Animals in Laboratories
   Use of animals outside of the centralized DLAM facilities
    is not permitted without IACUC approval.
    ◦ Laboratory Use – locations that are approved for animal use of short
      duration. Rats and mice may be taken to a laboratory for no more
      than 24 hours. USDA covered species may be taken to a laboratory
      for no more than 12 hours.
    ◦ Satellite Housing – locations approved for housing of the animals
      over 24 hours outside of the centralized facilities. Additional facility
      and husbandry requirements must be ensured.
 For any use of animals outside of the DLAM facilities,
  investigators must provide justification to the IACUC in a
  protocol or amendment.
 Satellite housing and areas where survival surgery is
  conducted must be approved by the IACUC prior to use.
 Areas approved for animal use need to be inspected by the
  IACUC semiannually.
         IACUC Approval Verification for
              Funding Agencies
    It is non-compliant to provide an IACUC date
       without obtaining a grant verification letter.
 IACUC must verify that animal use in a NIH (or other)
  funding proposal is the same as in the approved IACUC
  protocol(s) prior to release of funding.
 Submit the Funding Proposal Request Form and the
  proposal to the IACUC staff for review.
 If the proposal includes procedures not included in the
  IACUC protocol, amendments may be needed. Please
  allow enough time for this process to take place, if
  necessary.
                     Training Programs
   Approval to work with animals is contingent upon compliance with
    the training requirement. For new investigators, Mandatory Animal
    Care and Use (MACU) training is required and a 90-day grace period
    is provided. MACU training must be renewed every 3 years.
   Hands-on training is available from DLAM (eg. handling and restraint,
    injection techniques and gavage, blood collection techniques, and
    surgery).
   DLAM maintains a library of audiovisual and text references and
    gives DLAM Seminar Series talks on specific topics.
   Principal Investigators must keep in mind that it is their responsibility
    to guarantee that their students, associates, and staff are appropriately
    trained for the in vivo work in which they are involved. If additional or
    more specialized training is required, it can be provided through the
    DLAM veterinary staff.
Division of Laboratory Animal Medicine
       (DLAM) Services Provided
   Consultation on the selection, purchase, and use of
    animals
   Technical procedures
   Breeding colony organization and genotyping
   Receipt, transport, and housing of animals
   Daily observations of animal health
   Euthanasia
   Decontamination program
   Vermin control and eradication
    DLAM Centralized Animal Facilities
   Boston
    - Stearns/Arnold/South Cove 3rd and 4th floors
    - Ziskind 3rd floor & Surgical Research Laboratory
    - Transgenic Core in Tupper Building
   Medford
    - Bacon Hall
    - Dana/Barnum
             Surgical Research Laboratory
•   Provides dedicated operating rooms and facilities for large
    animal surgery
•   Space shared with DLAM for large animal housing


                Transgenic Core Facility
• Provides consultation for DNA preparation and embryonic
  stem (ES) cell tissue culture
 Produces transgenic and chimeric animals using DNA
  constructs and ES cells
 Provides rederivation services
 Provides cryopreservation
    Security Access into DLAM Facilities
   IACUC authorization is required for entry into animal facilities. In
    order to request access, please email Terri Naumes
    (terri.naumes@tufts.edu)
    ◦ Personnel must be added onto a protocol prior to facility access.
    ◦ Occupational Health Clearance must be obtained prior to facility access.
   On the Boston Campus (Stearns/Arnold/South Cove & Ziskind
    buildings), the perimeter is secured at both the main entrances and at
    each individual rooms by an swipe card system managed by the Tufts
    Police. This is your Tufts ID card that will be authorized with your
    specific room number(s).
   Visitors are not permitted in DLAM facilities for the protection of
    personnel, animals, and facilities.
   DO NOT SHARE YOUR ID CARD and don’t let others piggyback
    into the facility behind you.
  Occupational Health and Safety (OHS)
Working    with animals can expose research personnel to
potential hazards (such as bites, scratches or zoonotic diseases). The
PI is ultimately responsible for ensuring his/her staff obtain
the proper training about hazards associated with their
specific research.
  OHS is a mandatory item for compliance with the
   institution’s PHS Assurance and AAALAC accreditation
  New personnel must complete “Assessment Form for
   Animal Users”
  Contact Employee Health at 636-5480
  OHS Clearance requires annual renewals
Bite/Scratch Injury in the Animal Facility
If you are bitten or scratched by an animal or if you have been cut or
   scratched by an instrument or piece of equipment that has been
   exposed to an animal or its bodily fluids, then follow the basic first aid
   precautions immediately:
 Scrub the wound with antiseptic soap and flush the injured area
   liberally with water
 Contact the Occupational Health and Safety office and describe the
   incident. Follow their instructions regarding treatment and/or proceed
   directly to the treatment center.


  PROMPTLY REPORT ALL SUSPECTED WORK
  RELATED INJURIES AND ILLNESSES TO A DLAM
  SUPERVISOR AND YOUR SUPERVISOR
           Emergency Procedures
 In the event of an animal emergency, please
  contact a DLAM staff member for assistance
 For after-hours, contact Tufts Police and request
  that the veterinarian on-call be paged
 For personal injuries, contact a DLAM staff
  member and/or Tufts Police for assistance
 Emergency numbers are listed by phones in the
  facilities
    Personal Conduct in Animal Rooms
   Minimum Personal Protective Equipment (PPE)
    requirements
     disposable gowns
     Gloves
     Other PPE as directed on room door
 No eating, drinking, or loud music permitted
 Courtesy, Cleanliness, Awareness


             Promote a collegial atmosphere
                Procedure rooms
 In Boston each animal housing room has an assigned
  procedure room. Please do not take your animals to any
  procedure room other than your own.
 Sign up sheets are provided, otherwise space is on a first-
  come first-serve basis.
 Procedure rooms each include a bench top anesthesia unit,
  glass bead sterilizer & scale. Items removed from
  procedure rooms will not be replaced.
 Usage of isoflurane must be tracked. Mark the sheet next
  to the machine to document use. PLEASE contact DLAM
  when the isoflurane is out or is low, so that it can replaced.
      Individually Ventilated Caging
 Available in DLAM facilities on the Boston
  campus
 Provides HEPA filtered ventilation to each
  individual cage
 Provides a better environment for animals
 Provides higher level of protection from cross-
  contamination
               Biohazardous Agents
 All biohazardous agents must be approved by the
  Institutional Biosafety Committee prior to use.
 The IBC website can be found at:
http://www.tufts.edu/central/research/IBC/
 Biohazard card must be used for all biohazardous
  agents used in animals.
 These agents must be also be approved as part of
  the animal use protocol form prior to use.
 Investigators are mandated to follow the disposal
  directions given by the Environmental Health and
  Safety (EHS) office for these agents.
                             Chemical Hazards
Chemicals considered hazardous need to be included in an
  approved IACUC protocol and be registered with Tufts
  University EHS.
Please see the chemical hygiene page under the chemical safety
   link on the EHS web page at http://publicsafety.tufts.edu/ehs to
   determine what chemicals are considered hazardous.
A DLAM Safety Plan will be created by TU/TMC EHS for the use
   of each hazardous chemical in animals after it has been
   registered.
Prior to using a hazardous chemical, a meeting with a DLAM
   husbandry supervisor or manager needs to take place.
USE OF THE CHEMICAL CANNOT BEGIN UNTIL THE
  ABOVE HAS BEEN COMPLETED.
Chemical hazard card must be used for all chemical hazards used
  in animals.
All chemical waste collection supplies are available through
   TU/TMC EHS. It is the investigative staff’s responsibility to
   arrange for the delivery of these items and the pick up when the
   chemical use has been completed.
              Health Physics Approval
   All studies involving radioisotopes must be approved by
    Radiation Safety officer; DLAM; and the TMC radiation
    committee or Tufts University Radiation Hazards Control
    Group.
   No animal work involving radioisotopes is allowed until
    approval from the IACUC and consent of Health Physics has
    been obtained.
   Users must receive training in safe handling of radioisotopes
    prior to use.
   Failure to strictly comply with the requirements established by
    Health Physics and/or DLAM can result in substantial fines
    and/or sanctions.
           Disposal of Dead Animals
   All rodents found dead in the cage will be held in
    refrigeration for 3 days, after which they will be moved to
    a freezer.
    ◦ Dead animal stickers will be placed on the cage card holder of cages
      where dead animals were removed by DLAM.
    ◦ If a whole cage of animals is found dead by DLAM, the cage cards
      will be placed in a cardboard box located on the DLAM cage card
      supply rack in the facility (Boston campus).

   All animal carcasses and animal parts are to be disposed of
    in DLAM’s dead animal freezers.
                     Sentinel Program
   Sentinel animals are used to monitor the
    health status of rodent colonies.
   Cages are placed on the bottom of each
    rack and monitor the experimental
    animals housed on that rack.
   Please do not move these cages from their
    location on the racks.
   Diagnostic specimens are obtained from
    sentinels on a routine basis.
   Bimonthly, Semiannual, and Annual
    testing of Sentinel animals is conducted
    by DLAM.
   Investigators will be immediately notified
    if pathogen(s) are found in the colony.
   Yearly summary report is provided to
    investigators in the Fall.
 Infections in Research Animals
 Are   often subclinical (this means you cannot
 tell there is a problem by visual inspection)
 Can inhibit induction of animal model
 Can increase difficulty interpreting results
 Can increase variation in group (requiring
 larger sample sizes)
 Can have    zoonotic potential (meaning they
 can be transmitted from animals to people)
    Testing of Biological Products
 Transplantable tumors, hybridomas, cell lines,
  blood products, and other biologic materials can
  be sources of both murine and human viruses,
  which can contaminate rodents or pose risks to
  laboratory personnel.
 ALL biological products must be tested by
  DLAM prior to use (MAP, PCR, etc.). Even
  products (blood, serum) that have been passaged
  in rodents.
               Animal Procurement
 Animals must be covered by an IACUC protocol prior to
  ordering.
 Animals must be purchased through DLAM using only
  Approved vendors. Fill out the “DLAM Animal Purchase
  Request Form” and submit to DLAM.
 Animal procurement from Non-approved Vendors will
  require submission of request and approval from DLAM
  Director.
    ◦ Fill out the “Tufts Rodent Importation Request Form” and
      “DLAM Animal Purchase Request Form” and submit to
      DLAM.
    ◦ “Rodent Health Report for Importation Form” is sent to the
      non-approved vendor/institution and should be completed by
      that facility veterinarian or designated facility staff and
      returned to DLAM.
           Importation of Rodents from
             Non-Approved Sources
 Dependent upon clean health certificate received
  in advance of the shipment.
 Animals will be quarantined or can be rederived
  in the Transgenic Core.
    ◦ Quarantine is the separation of newly received
      animals from those already in the facility in order to
      prevent potential spread of contaminants until their
      health status has been determined, typically 10-12
      weeks.
    ◦ Rederivation has benefits of decreased time,
      availability of F1 progeny, and virtual elimination of
      pathogens including opportunistic organisms.
    Animal Arrival and Acclimation
 All animals should be afforded a period of acclimation/
  stabilization following shipment.
 Stabilization periods vary depending on species, type of
  transportation, and intended use of animal:
  ◦ Rodents: 2-3 days
  ◦ Large animals:
      Chronic procedure: 5 days
      Acute (< 2 weeks): 3 days
 Studies involving immune response, hormone levels, or
  response to stimulus should be given 72 hours to stabilize.
  ◦ Corticosterone levels & immune function affected by
     shipping
         Exportation of Animals
 DLAM can assist investigators who are shipping
  animals to other institutions.
 The receiving institution will require a health
  certificate from a DLAM veterinarian.
 Complete “Request for Health Certificate for
  Shipping Animals” and send to dlam-
  export@tufts.edu or fax to 6-8354.
 Do NOT ship animals until the receiving
  institution has provided approval of shipment.
Intra-Institutional Transportation of Animals

  See Intra-Institutional Policy on Transport of
    Animals from the Animal Facility to
    Laboratories
    • Transport animals in a new clean cage covered with an
      opaque drape (please don’t use yellow gowns) or in brown
      bags provided in facility for transport of rodent cages.
    • Rodent caging must be complete with filter tops.
    • Remove water bottles or reposition to prevent water
      leakage into cages.
    • Transfer rodents into another clean cage before returning to
      animal room.
Animal Protocol Transfers & Room Relocation
 Animals may be transferred from one protocol to another.
  Use the Animal Protocol Transfer Form to list the details
  and submit it to DLAM for Approval. The IACUC office
  will send an Approval email when all conditions of transfer
  have been Approved. Transfer should occur only after the
  IACUC office has emailed the Approval.
 If you need to move your animals into a different room,
  you will need to obtain approval from DLAM. You can do
  this by submitting the Animal Relocation Request Form to
  the DLAM office. Animals should not be moved until
  after you receive approval from DLAM.
 Both forms can be submitted to DLAM at fax #
 (617) 636-8354.
               Escaped Animals
 If an animal escapes in an animal room or your
  laboratory, please contact DLAM for assistance.
 Lab rodents caught in traps or found loose in
  animal rooms should be euthanized and may be
  necropsied by DLAM.
Animal Room Maintenance and Caging
 DLAM Husbandry staff performs daily
  maintenance of all animals, including routine
  cage changing.
 If investigative staff only want to provide care,
  then use “Do Not Disturb” cage card.
 Room racks have a limited supply of cages and
  equipment. Hallway racks have additional
  caging supplies.
 If caging is not available, contact the Husbandry
  office.
  DO NOT DISTURB Cage Card
 See “DLAM Policy Regarding           Investigative staff is
Animal Husbandry Procedures            responsible for placing and
Provided by Investigative Staff”       removing DND cage card.
                                      Assumes responsibility for
                                       all husbandry (cage change,
                                       food, water) and must
                                       maintain same frequency as
                                       DLAM.
                                      Date of last cage change
                                       must be provided and cages
                                       must be changed at least
                                       once a week.
                                      Lab staff must date and
                                       initial when cages are
                                       attended to.
     Special Husbandry Services

 Special husbandry by DLAM is available upon
  request or is done by the investigative staff.
 “Special Husbandry” cage card is used to identify
  cages for:
  ◦ Special food (eg: breeder chow)
  ◦ Treated water (eg: Tylenol)
  ◦ Special cage environment (eg: “no nestlets”)
Special Husbandry Cage Card
                 Placed and removed
                  by research
                  personnel
                 Card needs to be
                  filled out completely
                 If DLAM is asked to
                  provide special
                  husbandry you must
                  notify DLAM
                  Facility Supervisors
                  to coordinate.
            Animal Identification
 See IACUC Policy and Procedures for Animal
  Identification and Procedures for Corrective Action
 A cage card must be established for each animal and/or
  cage of group-housed rodents. Blank cards are available in
  the animal facilities.
 Investigators may submit a request to DLAM for a
  designated number of computer-generated cage cards with
  all required information. Send Cage Card Request Form to
  Maggie Collum and allow five days to process.
 Animals/cages found with incorrect identification, will be
  flagged with a Non Compliant cage card.
Required Information on Cage Cards
   Investigator’s name
   Investigator’s phone numbers, both office and
    laboratory
   Approved and active protocol number
   Current account number
   Species, strain or stock (if applicable)
   USDA tag # or other identification (if applicable)
   Contact person (individual responsible for the
    animals) and extension
Non-Compliant Cage Card
               Placed by DLAM
                when required
                information is
                missing from cage
                card
               Gives a 30 day
                window to correct
                the non-compliant
                issue
Overcrowded Cages
         Mouse Cage Populations
See IACUC Policy for Maintaining Mouse Cage
  Populations
Small standard shoebox:
 Five adult animals; or
 One adult pair and one litter – pups must be weaned at 21
  days of age; or
 One adult female and one litter – pups must be weaned
  between 22-28 days of age, no post-partum estrus breeding
  allowed; or
 Two adult females, with 1-2 litters – no more than 10 pups
  in cage, must be weaned at 21 days
Overcrowded Cage Cards
       Placed by DLAM
       Given a specific deadline either
        3 or 7 days
       3 day deadline: you will receive
        email notification.
        7 day deadline: you will not
        receive notification.
       Failure to correct the situation
        results in DLAM separation of
        the cage and lab will be charged
        a new cage setup fee.
       OC cage card can only be
        removed by DLAM staff.
      Inventory and Monthly Billing
 DLAM takes census once per week and records numbers
  on each room’s Census Sheet.
 Census Sheets are located near the entrance to each
  housing room.
 Use these sheets to recorded the number of cages added to
  or removed from the housing room so that you are charged
  per diem rates for the most accurate number of cages each
  day.
       Weaned/separated cages (+ the number of cages you generated)
       Euthanized cages (- the number of cages you removed)
   Monthly billing questions contact Betty Turner at
    (617) 636-5614.
    Daily Animal Health Evaluations
 Animal health is checked routinely by husbandry staff,
  veterinary technicians, and veterinarians.
 Investigator health check (HC) procedures are an
  integral part of program. Please contact DLAM and/or
  place a “Health Check” (HC) card to flag cages with
  health concerns.
 If a Health Check card has been placed on a cage, you
  can contact a vet tech (x6-2674) if you haven’t already
  been notified of the finding.
  ◦ Cards are placed to help monitor and track animal
     health and/or treatments
  ◦ Cards are only to be removed by DLAM staff
   Two sided card
       Front describes health
        concern(s) found
       Back is a treatment
        chart
   Card is dated and
    initialed by DLAM
    personnel to track
    treatments and
    progression
   Research personnel
    can record
    observations and/or
    sign off if they perform
    treatments.
      Room Logs
   Located on each animal
    room door
   Changed monthly
   Gives a quick overview
    of problems found in
    each room (such as
    overcrowded cages,
    health checks, dead
    animals found, etc.)
    during DLAM rounds
   Includes a key of
    abbreviations at the
    bottom of the form
      Technical Service Request Form
   Used by investigative staff to request DLAM services
   Forms can be found within the DLAM Animal Facilities, or
    call the DLAM office at X6-2674 and request blank form.
   Can request euthanasia, technical services (such as blood
    collection, injections, gavage, surgery, etc.), or training on
    various techniques.
   Please give 5 business days notice for scheduling with
    Veterinary Technicians.
                    Rodent Survival Surgery
 Survival surgery needs to be performed in a clean, dedicated space.
  For Boston campus investigators, please perform surgery ONLY in the
  procedure room associated with your animals’ housing room.
Gas anesthesia machines are available in every procedure room for
 isoflurane use. Please utilize the sign-up sheets to reserve space and the
 isoflurane sheets to track the time and use of the machine.
Aseptic technique is REQUIRED!
    -Disinfect surfaces prior to surgery
    -Equipment MUST be sterilized
    -A clean lab coat/disposable gown/ surgical scrub shirt, sterile surgical
    gloves, and a surgical mask all MUST be worn
    -Proper preparation of surgical site
    -Proper suture material and wound closure
 Use of the rodent “Surgery Card” on all cages is REQUIRED!
   Use of surgery card is required
    by IACUC policy
   It is the investigative staff’s
    responsibility to properly
    monitor animals in the post-op
    period for 3 days, which
    includes filling out the card
    completely.
   All relevant information on the
    card must be filled out, dated,
    and initialed each day.
   Improper monitoring or non-
    use is an issue of Non-
    Compliance.
       Post-Operative Care for Surgeries
 Investigative staff is responsible for providing all animals
  involved in survival surgical procedures appropriate post-
  op care, as described in the IACUC policy.
 Records of post-op care must be accurately maintained
 Animals must be kept warm during procedure and in the
  immediate post-op period while recovering from
  anesthesia. Electrical heating pads must not be used as they
    frequently develop “hot spots,” which can cause thermal burns.
   Continuous monitoring is required until animals have
    recovered consciousness and can maintain sternal
    recumbancy.
       Post-Operative Care for Surgeries
   Assess hydration/ fluid replacement/adequate nutrition (see Surgery
    policy for details).
   The incision must be examined for evidence of wound dehiscence or
    infection until it is completely healed and wound clips/sutures must be
    removed 10-14 days post-operatively.
   Analgesics, in general:
     ◦ Major operative procedures will require the administration of
       analgesics for 3 days post-operatively.
     ◦ For less invasive surgeries, frequency and duration of analgesic
       administration will depend on the type of surgery, but the minimum
       analgesia required is typically at least once peri-operatively or
       immediate post-operatively, unless scientific justification can be
       provided for not using all classes of analgesics.
             Animal Health Records
   Appropriate records must be kept by the investigative staff
    in order to document post-operative care evaluation and
    treatments. Please consult the IACUC Policy on
    Maintaining Proper Animal Health Records.
    ◦ For rodents, this information must be recorded on the blue Surgery
      cage card.
    ◦ For USDA covered animals; all procedures, including surgery,
      must be detailed in each animal’s medical record.


    Records are subject to inspection by IACUC,
            USDA, PHS, and AAALAC
Use of Anesthetics and Analgesics
 Adequate anesthesia and analgesics must be
  used to minimize pain and distress, unless
  justified in the approved protocol.
 NO EXPIRED or UNLABELED substances can
  be used in animals
 Chloroform is strictly prohibited – toxic to some
  mice and suspected carcinogen
 Ether is strictly prohibited –explosive!
     Ordering Controlled Substances
   Controlled substances can be ordered through DLAM.
   Use of the specific drug must be included in an approved
    protocol.
   All controlled drugs must be kept behind two locks.
   A usage log including; drug name, amount used for each
    administration and date, amount remaining, and user ID
    must be maintained and is subject to inspection.
   The “Authorization for the Acquisition of Controlled
    Substances” form is filled out by PI granting permission for
    lab personnel to request controlled substances from DLAM.
   Fill out and submit a “DLAM Drug and Supply Request
    Form” each time you wish to order drugs or supplies from
    DLAM.
    - Please allow 5 business days
    - Some items may take longer to order, so plan accordingly.
         Policy on Protocol Non-Compliance and/or
                    Animal Mistreatment
Situations of noncompliance can vary considerably and the IACUC is required to
determine the significance of each issue. Examples are listed below:
   Minor issues are typically handled at the institutional level (unless there are repeat violations).
    Examples may include, but are not limited to:
    ◦ Lack of the mandated surgical and post-operative care records
    ◦ Failure to adhere to proper aseptic technique for survival surgery
    ◦ Not informing the IACUC of the addition of personnel
    ◦ Unapproved transfer of animals from one protocol to another

   Significant issues that are deemed serious must be reported to the Office of Laboratory Animal
    Welfare (OLAW) at the National Institutes of Health (NIH) and/or to the United States
    Department of Agriculture (USDA), as well as reported to the specific funding agency for that
    project. Examples may include, but are not limited to:
    ◦ Acquiring animals for research or performing unapproved procedures without IACUC
       approval
    ◦ Performing a procedure in such a manner that animals endure distress, pain, or suffering
       that is not addressed in the approved protocol.
    ◦ Repeated incidents of minor issues of non-compliance.
    The IACUC is mandated to comply with the following regarding how to classify non-
       compliance, which is explained in the OLAW notice found here:
       http://grants.nih.gov/grants/guide/notice-files/NOT-OD-05-034.html
                        Euthanasia
   The method of euthanasia described in your protocol must
    comply with the Report of the American Veterinary
    Medical Association’s Panel on Euthanasia. For example:

    Approved methods include:
    ◦ CO2 asphyxiation, which must be followed by a
      physical method to ensure death. CO2 must be given via
      a gas cylinder. Use of dry ice is NOT permitted.
    ◦ Anesthetic overdose
    ◦ General anesthesia followed by perfusion, removal of
      major organ, or cervical dislocation.

    Conditional methods require scientific justification in the
     protocol. These include methods such as decapitation or
     cervical dislocation while the animal is conscious.
Euthanasia Rack

   Animals left on the rack for euthanasia must be afforded the same
    conditions as all animals housed in this facility:
   Do not house more than 5 adults per cage
   Do not leave pre-weaned litter w/o their mother
   Do not stack cages on top of each other
   Do not leave animals w/o adequate food and water
   DO contact DLAM immediately if you see any of the above
    conditions

Individuals who do not comply with the above requirements risk
not only their suspension from work at this institution, but the
suspension of the protocols under which they are conducting
research.
This concludes the MACU Training power
point presentation.

Please continue on by reviewing the IACUC
Policy folder before taking the quiz.

          Thank you

				
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