Introduction to the Use of Animals and the DLAM Facilities by D928wS


									Mandatory Animal Care and Use (MACU)

     Introduction to the Use of Animals
          and the DTRR Facilities

   Institutional Animal Care and Use Committee

     Tufts University & Tufts Medical Center
    Why must you participate in this
          on-line training?
 You will be using or working with animals
  in research or teaching
 You need to have a basic understanding of
  the complex system that is meant to
  ensure the protection of animals
 We are obligated by law to make sure
  everyone using animals on campus is
  aware of the regulations
        Outline of Presentation

   Introduction to Federal and State Regulations
   Institutional Animal Care and Use Committee
   Division of Teaching and Research Resources
   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)
   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)
   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
   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
   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:
    Massachusetts Research Animal
 Massachusetts Research Animal Regulations were
  passed in 1985.
 Overseen by the MA Department of Public Health
 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.
    Association for the Assessment and Accreditation of
          Laboratory Animal Care, International

   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
AGENCY       REPORT                 INSPECTION
USDA APHIS   Annual                 Annual
AC           Registration 3 years   Unannounced

PHS OLAW     Annual                 Possible
             Assurance 4 years

MA DPH DFD   Annual                 Annual

AAALAC       Annual                 Site visit 3 years
             Program description
             every 3 years
          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
    - Elizabeth Prescott, IACUC/IBC Regulatory Coordinator
    - 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
   Attending veterinarian(s) (AV)
   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
   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) an issue of non-compliance is voted to be reportable
         by the IACUC
     2) all federal reports are submitted
     3) financial resources are necessary to maintain
        compliance in the program or the facilities.
              IACUC Protocol Form
 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
 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
           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
    without 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.
    ◦ There are some situations that will require Full Committee review,
      such as change in species, significant increase in animal numbers,
      additional Category E animals, or Category E procedure changes.
    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.
   The PI must submit the Funding Proposal Request Form
    and the proposal to the IACUC staff for review to start the
   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
      Use of Animals in Laboratories
   Use of animals outside of the centralized DTRR 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 DTRR 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.
         Security Access into DTRR
 IACUC authorization is required for entry into animal facilities
  – OHS Clearance and addition onto protocol must be completed
  prior to facility access
 Keycard and key access to buildings
 Key access to animal rooms
    ◦ Everyone must have their own KEYCARD/KEY.
       Get your own keycard/key when you start at TCSVM.
       Return your keycard/key to the police when your employment at TCSVM ends.
  ◦ Don’t let others piggyback into the facility behind you
 Access request form can be obtained at the link below:
  ◦ Send directly to Dr. George Lathrop or Dr. Lara Helwig,
    Building 17
Division of Teaching and Research Resources
         (DTRR) Services Provided
   Consultation on the selection, purchase, and use of
   Technical services
   Staff training
   Receipt, transport, and housing of animals
   Daily observations of animal health
   Euthanasia
   Decontamination program
   Vermin control and eradication
                      Training Programs
   Approval to work with animals is contingent upon compliance with the training
    requirement which you are doing now. 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 available from DTRR
    ◦ Mouse and rat handling
       Restraint and handling
       Injection techniques
       Blood collection techniques
    ◦ Rodent surgery
       NIH training video
       Models available
       Required for everyone doing surgery in rodents
    ◦ Cage changing (coming soon)
       Will be a required session
   DTRR maintains library of audiovisual and text references.
   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 DTRR veterinary staff.
             DTRR Facilities

           AAALAC accredited since 1997
Housing for all research species (small mammals
  and livestock species)
3 Surgical suites
  2-with HEPA filtration
  1-equipped with a Biological Safety Cabinet
Several ABL-2 housing and research suites
Several shared procedure rooms equipped with
  Chemical Fume Hoods
RBL-Regional Biosafety Laboratory
          Animal Manipulations
   Procedure Rooms
   Bldg. 20
    ◦ Room 009 – chemical fume hood, sink, table, C02
    ◦ Room 004 – anesthesia machine, tables
   RBL
    ◦ Procedure Rooms 1442, 1453, 1454, 1522, 1523,
      1532, 1533, 1542, 1543, 1552 and 1553

   Animals must be alert and mobile before being
    returned to housing rooms
        Caging Supplies and
        Equipment Sanitation
• Caging supplies and water bottles can be
  found in Bldg. 20, Room 013 Clean Side
• Sterile caging supplies can be found in
  Bldg. 20, Room 013
• If you need large numbers of cages on a
  certain day, please sign up using the
  calendars in the corridor of Bldg. 20,
  outside room 013
 RBL – Clean Cage Wash Room 1401
Personal Conduct in Animal Rooms
   Minimum Personal Protective Equipment (PPE)
     Disposable Gowns
     Gloves
     Other PPE as directed on room door
 No eating, drinking, or loud music permitted
 Courtesy, Cleanliness, Awareness

            Promote a collegial atmosphere
  Bite/Scratch Injury in the Animal
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 your institutional health care provider and describe the
   incident. Follow their instructions regarding treatment and/or proceed
   directly to the treatment center.

        Emergency Procedures
 In the event of an animal emergency, please
  contact a DTRR staff member for assistance
 For after-hours, contact Tufts Police and request
  that the veterinarian on-call be paged
 For personal injuries, contact a DTRR staff
  member and/or Tufts Police for assistance
 Emergency numbers are listed by phones in the
     Occupational Health and Safety
Working   with animals can expose research personnel to
potential hazards (such as bites, scratches or zoonotic diseases). The
PI is ultimately responsible for ensuring their staff obtains the
proper training about hazards associated with their specific
  OHS is a mandatory item for compliance with the
   institutions PHS Assurance and AAALAC accreditation
  New personnel should complete the “Health
  OHS Clearance requires annual renewals
  Contact:
    ◦ Milda Walkley, RN, 1 Ridge Road (Building 17)
      phone: 508- 251-7260
            Biohazardous Agents
 All biohazardous agents must be approved by the
  Institutional Biosafety Committee prior to use.
 The IBC website can be found at:

 All biohazardous agents used in animals must be
  listed on the cage card.
 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 EHS for these agents.
              Chemical Hazards
 All chemical hazards must be approved by the
  Environmental Health and Safety office prior to
 The EHS website can be found at:

 All chemical hazards used in animals must be
  listed on the cage card.
 These agents must also be approved as part of the
  animal use protocol form prior to use.
 Investigators are mandated to follow the disposal
  directions given by EHS for these agents.
         Health Physics Approval
 All studies involving radioisotopes must be
  approved by Radiation Safety officer; DTRR; and
  the Tufts University Radiation Hazards Control
 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 DTRR can
  result in substantial fines and/or sanctions.
       Disposal of Dead Animals
 All animal carcasses, and animal parts are to be
  disposed of in the DTRR freezers
  ◦ Tunnel between Bldgs. 21 and 22
 All animals from the RBL will go into tissue
 If you are using larger animals and require
  disposal drums, please contact DTRR
 NOTE: Unmanipulated rodents may be donated
  to the wildlife clinic for feeding purposes. Only if:
  ◦ No drugs/implants have been used
  ◦ Euthanasia via CO2 asphyxiation/physical
  ◦ They are not transgenic
                      Sentinel Program
   Sentinel animals are used to monitor the health status of rodent
   Please do not move these cages from their location on the racks and
    do not remove sentinels from the room at any time.
   Diagnostic specimens are obtained from sentinels on a routine basis
    by DTRR.
   Sentinel program is essential for disease tracking.
   Investigators will be immediately notified if pathogen(s) are found in
    the colony.
 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 (this means
    they can be transmitted from animals to people)
      Testing of Biological Products
   Biological products are the most common sources of
    introduction of infectious disease in a colony.
   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
   ALL biological products (cells, tumor lines, etc.) must
    be tested prior to use (MAP, PCR, etc.) and the
    results sent to DTRR. Contact Dr. Lara Helwig for
    more information.
   Includes products (blood, serum) that have been
    passaged in rodents.
               Animal Procurement
   Animals must be covered by an IACUC protocol prior to
   Animals must be purchased through DTRR using only
    Approved Vendors. DTRR maintains an Approved Vendor
    List that is frequently updated.
   Fill out the “Request for Laboratory Animal Purchase and
    Care” and submit to DTRR.
   Animal procurement from Non-Approved Vendors will
    require submission of request and approval from the
    DTRR Director.
    ◦ A health certificate must be received in advance of the
    ◦ If animals do not meet the health criteria for housing in
      DTRR facilities, off-site quarantine or rederivation may be
     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
    ◦ Rabbits: 1 to 2 weeks
    ◦ Large animals:
        Chronic procedure: 5 days
        Acute (< 2 weeks): 3 days
   Exceptions may be made with veterinary approval.
   Studies involving immune response, hormone levels, or
    response to stimulus should be given 72 hours to stabilize.
    ◦ Corticosterone levels & immune function affected by
         Exportation of Animals
 DTRR can assist investigators who are shipping
  animals to other institutions.
 The receiving institution will require a health
  certificate from a DTRR veterinarian.
 Do NOT ship animals until the receiving
  notification that the receiving institution has
  provided approval of shipment.
     Transportation of Rodents
           within DTRR
See Intra-Institutional Policy on Transport of Animals from
  the Animal Facility to Laboratories
    ◦ Rodent caging must be complete with filter covers, when
    ◦ Remove water bottles or reposition to prevent water leakage into
    ◦ Individual cages may be transported by hand within a building
    ◦ For transport of multiple cages, please use a cart
    ◦ Use indoor routes of travel (e.g., use tunnels and elevators)
    ◦ Carts may be used but should be disinfected (use Quat PV bottles
      available in the corridor) before returning to the animal facility
    ◦ If large numbers of cages must be moved, plastic containers can
      be used as a secondary containment system
    ◦ Cages should not be carried by hand outside without a secondary
      containment device (e.g., a bag or container)
Transportation of Large Animals
   Rabbits and other larger, non-rodent
    animals can be transported in Vari-kennel
    enclosures which are available from
   Transport of large animals (sheep, pigs)
    between campus buildings can be done
    by DTRR (using the Request for Services Form)
   The Transport Devices must be cleaned
    after each use and disinfected on a regular
               Escaped Animals
 If an animal escapes in an animal room or your
  laboratory, please contact DTRR for assistance.
 Lab rodents caught in traps or found loose in
  animal rooms should be euthanized and may be
  necropsied by DTRR.
       Special Husbandry Services
 Special   husbandry by DTRR is
       available upon request.
    ◦ Special food
    ◦ Special cage environment
      Such as “No nestlets”
    Special Husbandry Cards are made
    specific to the PI request and placed on
    the animal cages.
            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.
   Initially generated by animal order form
   If you need additional cage cards, contact
    Sherry Castonguay at 8-4981
 Animals/cages found with incorrect identification, DTRR
  contacts the PI to make any corrections.
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
   Emergency contact name and phone number
   Biohazardous or radioactive agent use
Cage Card – example 1
Cage Card – example 2
Overcrowded Cages
         Mouse Cage Populations
See IACUC Policy for Maintaining Mouse Cage
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. Pups must be within
  1-2 days of each other in age.
      Inventory and Monthly Billing
 DTRR takes census routinely 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
       Weaned/separated cages (+ the number of cages you generated)
       Euthanized cages (- the number of cages you removed)
   Monthly billing questions contact Susan Weidman
    at x-84642 or .
    Daily Animal Health Evaluations
 Animal health is checked daily by husbandry staff, vet
  tech and DTRR veterinarians.
 Investigator health check procedures are an integral part
  of program.
  ◦ Please contact DTRR and/or place “Problem
     Notification” card to flag cages with health concerns.
 If a “Problem Notification” card has been placed on a
  cage, you can contact DTRR, 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 DTRR staff.
Problem Notification Card
  Problem Notification Cards
Single sided card
Front lists
   Common problems
Dated  and initialed by DTRR personnel to
track treatments and progression
Research personnel may sign it if performing
May only be removed by DTRR
 Request for DTRR Services Form

Use to:
  Schedule surgery suites and procedure
  rooms (except 20-009)
  Order pharmaceuticals or other supplies
  Request technical assistance in
  Request animal transport
 Submit via fax to x87996
               Rodent Survival Surgery
 Rodent Surgery Training is required for all who will be performing
Rodent Surgery. Contact Dr. Lara Helwig for more information.
Survival surgery needs to be performed in a clean, dedicated
Gas anesthesia machines are available in every procedure room
for Isoflurane use. Use the sign up sheets to reserve space and
Isoflurane sheets to track the time in which the Isoflurane machine
was in use.
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
         Proper preparation of surgical site
         Proper suture material and wound closure
   Use of Rodent Postoperative Monitoring card is required
    by IACUC policy.
   It is the Investigators staff’s responsibility to properly
    monitor animals in the post-op period for 3 days, which
    includes filling out the monitoring card completely.
   All animal observations and analgesics given must be
   Improper monitoring is an issue of Non-Compliance.
             Post-Operative Care
 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.
  ◦ Cage Cards (for Rodents)
  ◦ Individual Animal Records (for USDA covered species)
 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
   Assess hydration/ fluid replacement/adequate nutrition (see policy for
   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 postoperative care evaluation and
    treatments. Please consult the IACUC Policy on
    Maintaining Proper Animal Health Records.
    ◦ For rodents, this information must be recorded on the Cage Cards.
    ◦ For USDA covered animals, all procedures, including surgery,
      must be detailed in each individual animal’s medical record.

    Records are subject to inspection by IACUC,
            USDA, PHS, and AAALAC
Animal Health Records
              Animal health records must be
               complete, including:
              All procedures (performed by
               either lab personnel or DTRR)
              Injections (drug, concentration,
               volume, site/route)
              Blood collection (volume, site)
              Other manipulations (must be in
               approved IACUC protocol)
              Observations
              Treatments
              Adverse events and outcomes
              Surgery reports and post-op care
Anesthetics, Analgesics, and Tranquilizers

    Adequate anesthesia, analgesics, and
     tranquilizers must be used to minimize pain and
     distress, unless justified in the approved
    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 DTRR.
   Their use must be included in an approved protocol.
   You must maintain a drug log of usage and all controlled
    drugs must be kept behind two locks.
   The “Authorization for the Acquisition of Controlled
    Substances” form is filled out by PI granting permission for
    lab personnel to request controlled substances from DTRR.
    This form is filled out one time only, and kept on file in
    DTRR for reference.
   Fill out and submit a “DTRR Request for Services Form”
    each time you wish to order drugs or supplies from DTRR.
        Please allow 5 business days
        Some items may take longer to order, so plan
    Policy on Protocol Non-Compliance and/or
               Animal Mistreatment
Situations of Noncompliance can vary considerably and determination of significance
is made on a case-by-case basis based on the particular circumstances surrounding the
   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:
 AVMA Panel of Euthanasia should include the
  method of euthanasia described in your protocol.
  ◦ Approved methods (CO2, anesthetic OD)
  ◦ Conditional methods require justification
    (decapitation, cervical dislocation).
 CO2 euthanasia must be performed via gas
 Authorization (on protocol) required to perform
          When Euthanizing Large
            Groups of Rodents
   Animals 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 not mix animals from cages.
   DO contact DTRR immediately if you see any of the above

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
This concludes the MACU Training power
point presentation.

Please continue on by reviewing the IACUC
Policy & DTRR/DLAM Guideline folders
before taking the quiz.

          Thank you

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