LATG: A Review of The Guide for the Care and Use of Laboratory Animals

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LATGA Review ofThe Guide for the Care and Use
of Laboratory Animals

• Created by
• Marc S. Hulin, DVM, Dipl. ACLAM

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History of The Guide

• First published in 1963 6th revision
• Current revision was written by ILAR
• Purpose assist institutions in caring for and
  using animals in ways judged to be
  scientifically, technically, and humanely
  appropriate
• 6th edition was supported by NIH, USDA, and Dept.
  VA

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Introduction

• Laboratory Animal any vertebrate animal used in
  research, teaching or testing
• Guide endorses 9 US Government principles, AWA,
  PHS policy, other federal and state laws

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Evaluation Criteria

• Performance standards preferred over Engineering
  standards
• performance define an outcome in detail and
  provide criteria for assessing the outcome, but
  dont limit methods to achieve outcome
• Must considered imperative
• Should strong recommendation

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I. Institutional Policies and Responsibilities

• IACUC membership
• DVM with lab animal experience
• 1 practicing scientist with research experience
  with animals
• 1 public member (non-affiliated)
• Physical restraint
• use of manual or mechanical means to limit
  animals movement for examination, collection of
  samples, or drug administration

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Physical Restraint

• Prolonged restraint should be avoided unless it
  is essential for research objectives and approved
  by IACUC
• Guidelines
• not to be considered normal methods of housing
• restraint period should be minimum time
• animal should be trained
• regular observations
• veterinary care provided if lesions or illness

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Multiple Major Surgical Procedures

• Major surgery
• penetrates and exposes a body cavity or produces
  substantial impairment of physical or physiologic
  function
• Multiple major surgeries on a single animal are
  discouraged unless
• scientifically justified
• approved by the IACUC
• related components of a research project
• conserve scare animal resources

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Food or Fluid Restriction

• If experimental protocols require restriction at
  least minimal quantities of food and fluid should
  be available
• Restriction for research purposes should be
  scientifically justified
• Highly preferred food or fluid as positive
  reinforcement, instead of restriction, is
  recommended

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Veterinary Care

• Adequate veterinary care must be provided,
  including access to all animals for evaluation of
  their health and well-being
• Veterinarian must provide investigators with
  advice on use of sedatives, analgesics, or
  anesthetics in animals

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Personnel Qualification Training

• Personnel caring for animals should be
  appropriately trained and the institution should
  provide for formal or on-the-job training to
  facilitate effective implementation of the
  program and humane care and use of animals
• Occupational Health and Safety program must be
  part of the overall animal care and use program

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Preventative Medicine for Personnel

• Development and Implementation of a program for
  medical evaluation should involve input from
  trained health professionals
• Cercopithecine Herpesvirus 1 (Herpes B virus)
• personnel who work with Macaques should have
  access to and be instructed in the use of bite
  and scratch emergency kits

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Animal Environment, Housing, Management

• Proper housing and management
• Animal well-being
• Quality of research data
• Health and safety of personnel
• Animals should be housed with goal of maximizing
  species-specific behaviors and minimizing stress
  induced behavior

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Micro vs. Macroenvironment

• Microenvironment
• physical environment immediately surrounding it,
  i.e. primary enclosure, food, wire bar lid,
  sipper tube.
• Macroenvironment
• the physical environment of the secondary
  enclosure, such as a room, barn, or an outdoor
  habitat
• Linked by ventilation between primary and
  secondary enclosure

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Primary Enclosures

• Should be constructed with materials that balance
  the needs of the animal with the ability to
  provide sanitation
• Solid-bottom caging, with bedding, is recommended
  for rodents

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Space Recommendations

• Based on professional judgment and experience
• Thigmotactic rodent species benefit more from
  wall space (stay along the walls in cages)
• Animal must have enough space to turn around and
  express normal postural adjustments
• Social animals should be housed in pairs or groups

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Space Recommendations

• Cage height
• Mice 5 inches
• Rats 7 inches
• Hamsters 6 inches
• Guinea pigs 7 inches
• Rabbits 14 inches
• Cats 24 inches

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Space Recommendations

• Floor space
• Dogs
• lt15 kg 8.o sq. ft.
• 15-30 kg 12.0 sq. ft.
• gt 30 kg 24.0 sq. ft.
• Monkeys
• Groups 1-7 (including baboons)
• Apes (Pongidae) Groups 1-3
• Brachiating species cage height should be when
  fully extended, swing from cage ceiling without
  feet touching the floor

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Temperature and Humidity

• Regulation of body temperature within normal
  variation is necessary for the well-being of lab
  animals
• Relative humidity should be controlled to the
  acceptable range of 30-70

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Dry Bulb Temperatures

• Mouse, Rat, Hamster, Gerbil, Guinea Pig
• 64-790 F
• Rabbit
• 61-720 F
• Cat, Dog, NHP
• 64-840 F
• Farm animals Poultry
• 61-810 F

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Ventilation

• Supply adequate oxygen remove thermal loads
  caused by animals and equipment
• Use of computer modeling for designing facility
  ventilation (computational fluid dynamics)
• 10-15 fresh-air changes per hours for secondary
  enclosures is general standard
• Recycled vs. Non-recycled air

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Illumination

• Light can affect the physiology, morphology, and
  behavior of various animals
• Lighting should provide sufficient illumination
• for well-being of the animals
• allow good housekeeping practices
• Time-controlled lighting system should be used to
  ensure a regular diurnal cycle

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Illumination

• Light levels of 325 lux (30 ft-candles) about
  1.0m above the floor appear to be sufficient for
  animal care and do not cause signs of phototoxic
  retinopathy in albino rats
• Light at cage level for animal susceptible to
  phototoxic retinopathy should be between 130-325
  lux

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Noise

• Noisy animals (dogs, swine, etc.) should be
  housed away from quieter animals, such as
  rodents, rabbits, and cats
• Noise gt85 dB can have both auditory and
  non-auditory effects
• eosinopenia
• increased adrenal weights in rodents
• reduced fertility in rodents
• increased blood pressure in NHP

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Behavioral Management

• Structural Environment
• primary enclosure- cage complexities, cage
  furniture, manipulanda
• Social Environment
• physical contact and communication with
  conspecifics (members of same species)
• Activity
• animals should have opportunities to exhibit
  species-typical activity patterns

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Husbandry

• Food
• unused, opened bags of food should be stored in
  vermin-proof containers
• exposures to temp. gt 700 F and humidity extremes
  hastens deterioration
• dry lab animal diets stored for 6 months after
  manufacture
• Vitamin C diets- 3 months shelf-life
• Stabilized forms of Vitamin C extend shelf life

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Bedding

• Untreated softwood shavings and chips are
  contraindicated may affect animals metabolism
• Cedar shavings are not recommended, because
  microsomal enzymes and cytotoxicity
• Soiled bedding should be removed and replaced
  with fresh materials as often as is necessary to
  keep the animals clean and dry

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Cleaning and Disinfection of Primary Enclosures

• If animal waste is to be removed by flushing-at
  least once daily (animals kept dry during
  flushing)
• enclosures and accessories (tops) should be
  sanitized at least once every 2 weeks
• Solid-bottom caging, bottles, and sipper tubes
  usually require sanitization at least once a week

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Cleaning and Disinfection of Primary Enclosures

• Effective disinfection can be achieved with wash
  rinse water at 143-1800 F
• Traditional 1800 F for rinse water refers to the
  water in the tank or sprayer manifold
• A regularly scheduled and documented pest control
  and monitoring should be implemented

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Emergency, Weekend, and Holiday Care

• Animals should be cared for by qualified
  personnel every day, including weekends and
  holidays
• Emergency veterinary care should be available
  after work hrs., weekends, and holidays
• A disaster plan that takes into account both
  personnel and animals should be prepared as part
  of the overall safety plan for the animal facility

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Identification and Records

• Toe-clipping, for identification of small
  rodents
• only when no other method is feasible
• performed only on altricial rodents
• Clinical records for individual animals
• valuable for dogs, cats, NHP
• contain pertinent clinical and diagnostic info.
• dates of vaccinations, surgery, experimental use

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Veterinary Medical Care

• Adequate veterinary care consists of effective
  programs for
• Preventative Medicine
• Surveillance, Diagnosis, Treatment, and control
  of disease including Zoonosis
• Management of protocol associated disease
• Anesthesia and Analgesia
• Surgery and Postsurgical care
• Assessment of Animal Well-Being
• Euthanasia

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Veterinary Medical Care

• A veterinary care program is the responsibility
  of the attending veterinarian, who is certified
  or has training or experience in lab animal
  science and medicine
• The veterinarian must provide guidance to
  investigators and all personnel involved in the
  care and use of animals

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Animal Procurement Transportation

• All animals must be acquired lawfully
• Dogs and cats from Class B dealers should be
  carefully inspected for special identification
  markers
• Importation of NHP is regulated by PHS with
  specific guidelines for Tuberculin testing

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Quarantine, Stabilization, and Separation

• Effective quarantine program minimizes the chance
  for introduction pathogens into an established
  colony
• Veterinary Medical staff should have procedures
  for evaluating the health and pathogen status of
  newly received animals

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Quarantine, Stabilization, and Separation

• Effective quarantine program for NHP
• limit exposure of humans to zoonotic infections
• filoviral and mycobacterial infections in NHP
  have necessitated specific handling guidelines
  (CDC)
• Newly received animals should be given a period
  for physiologic, psychologic, and nutritional
  stabilization before their use.

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Quarantine, Stabilization, and Separation

• Physical separation of animals by species is
  recommended to prevent interspecies disease
  transmission and conflict
• separate rooms
• cubicles
• laminar-flow units
• cages that have filtered air or separate
  ventilation
• isolators

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Quarantine, Stabilization, and Separation

• Examples of need for separate housing by species
  
• Bordetella bronchiseptica in rabbits- severe
  disease in guinea pigs
• Simian Hemorrhagic Fever and SIV separate New
  World, Old World African, and Old World Asian
• Squirrel monkey latently infected with
  Herpesvirus tamarinus which is fatal to Owl
  Monkeys

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Surveillance, Diagnosis, Etc.

• All animals should be observed for signs of
  illness, injury, or abnormal behavior daily
• Unexpected deaths and signs of illness should be
  reported promptly for appropriate veterinary
  medical care
• The choice of medication or therapy should be
  made by the veterinarian in consultation with the
  investigator

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Surveillance, Diagnosis, Etc.

• Infectious agents that affect research
• Sendai, KRV, MHV, LCMV, and Mycoplasma pulmonis
• The principal method for detecting viral
  infections is serologic testing
• Transplantable tumors, hybridomas, cell lines,
  and other biologic materials should be tested for
  murine viruses
• MAP (mouse antibody production) test, RAP, HAP
  used for monitoring for viral contamination

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Surgery

• Appropriate attention to presurgical planning,
  personnel training, aseptic and surgical
  technique
• Use of antibiotics should never be considered as
  a replacement for aseptic procedures
• PHS policy and AWA place responsibility with the
  IACUC for determining that personnel are
  qualified and trained for surgery

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Surgery

• Major survival surgery
• penetrates and exposes a body cavity or produces
  substantial impairment of physical or physiologic
  function
• laparotomy, thoracotomy, craniotomy, joint
  replacement, limb amputation

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Surgery

• Minor survival surgery
• does not expose a body cavity and causes little
  or no physical impairment
• wound suturing, peripheral-vessel cannulation,
  castration, prolapse repair, skin biopsy
• most procedures routinely done on an outpatient
  basis in veterinary clinical practice

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Surgery

• Nonsurvival surgery
• animal is euthanized before recovery from
  anesthesia
• does not require aseptic surgery
• surgical site should be clipped, wear gloves, and
  clean instruments
• Skin sutures, wound clips, or staples
• must be removed in timely manner
• Veterinary medical standard 10-14 days

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Surgery

• Liquid chemical sterilants should be used with
  adequate contact times
• Alcohol is neither a sterilant nor a high-level
  disinfectant
• Nonrodent aseptic surgery should be conducted
  only in designated surgical facility
• PI and veterinarian share responsibility for
  ensuring that postsurgical care is appropriate

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Pain, Analgesia, and Anesthesia

• Fundamental to pain relief is ability to
  recognize clinical signs in specific species
• vocalization, depression, behavioral changes,
  immobility, abnormal posture
• Anthropomorphic concept
• Neuromuscular blocking drugs do not provide pain
  relief
• paralyzes muscle animal feels pain but unable to
  move

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Euthanasia

• The act of killing animals by methods that induce
  rapid unconsciousness and death without pain or
  distress
• Methods should be consistent with 2000 Report of
  the AVMA Panel on Euthanasia
• Avoid animal distress
• other animals should not be present when
  euthanasia is performed

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Construction Guidelines

• Corridors
• should be wide enough to facilitate the movement
  of personnel and equipment
• 6-8 ft. wide can accommodate most facilities
• Animal-Room Doors
• should open into animal rooms
• large enough for easy passage of racks and
  equipment
• 42 x 84 inches

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Construction Guidelines

• Floors
• moisture-resistant, nonabsorbent,
  impact-resistant, and smooth
• Drainage
• if used should be sloped and drain traps kept
  filled with liquid
• drainpipes should be at 4 inches in diameter
• floor drains are not essential in all animal rooms

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Construction Guidelines

• HVAC systems
• capable of adjustments or - 20 F
• relative humidity 30-70
• temperature is best regulated by having
  thermostatic control for each room
• regular monitoring of HVAC system done at room
  level
• partial redundancy

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Construction Guidelines

• Power Lighting
• in event of power failure an alternative or
  emergency power supply should be available to
  maintain critical services
• recessed energy-efficient fluorescent light is
  most commonly used
• time-controlled lighting system should be used to
  ensure a uniform diurnal lighting cycle

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Construction Guidelines

• Storage areas
• bedding and food should be stored in separate
  area from risk of contamination from toxic or
  hazardous
• refrigerated storage kept below 70 C to reduce
  putrefaction of wastes and animal carcasses

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Facilities for Aseptic Surgery

• Rodent surgery
• small simple dedicated space in lab minimize
  contamination during surgery
• Large animal surgery
• surgical support, animal preparation, surgeons
  scrub, operating room, and postoperative recovery

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