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

Created by

Marc S. Hulin, DVM, Dipl. ACLAM

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

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

Evaluation Criteria

Performance standards preferred over Engineering standards performance: define an outcome in detail and

provide criteria for assessing the outcome, but don’t limit methods to achieve outcome

“Must”: considered imperative “Should”: strong recommendation

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

animal’s movement for examination, collection of samples, or drug administration

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

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

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

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

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

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

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

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

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

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

Space Recommendations

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

Space Recommendations

Floor space: Dogs:

<15 kg: 8.o sq. ft. 15-30 kg: 12.0 sq. ft. > 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

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%

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

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

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

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

Noise

Noisy animals (dogs, swine, etc.) should be housed away from quieter animals, such as rodents, rabbits, and cats

Noise >85 dB can have both auditory and non-auditory effects: eosinopenia increased adrenal weights in rodents reduced fertility in rodents increased blood pressure in NHP

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

Husbandry

Food: unused, opened bags of food should be stored

in vermin-proof containers exposures to temp. > 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

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

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

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

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

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

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

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

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

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

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.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Facilities for Aseptic Surgery

Rodent surgery: small simple; dedicated space in lab; minimize

contamination during surgery Large animal surgery:

surgical support, animal preparation, surgeon’s scrub, operating room, and postoperative recovery