Laboratory Animal Care and Use Program and Facilities

Judy Hickman-Davis, DVM, PhD, DACLAM

ALAT Chapter 13 Colony Health surveillance

“Good research depends on the availability of

healthy research animals.”

Laboratory Animal Care and Use Program and Facilities

Health Surveillance

When does health surveillance begin? Before the animals are received Procurement form and health reports Quarantine and re-evaluation

Specific Selected vendors LAT receiving Follow up schedule

PE, vaccinations, testing, treatment

Laboratory Animal Care and Use Program and Facilities

Animal Observation

What is the most important factor in health surveillance and disease prevention? Husbandry LAT is the first line of defense Early detection is ESSENTIAL!

Laboratory Animal Care and Use Program and Facilities

Animal Observation

General Body Condition Stunted growth Weight loss Swelling (lumps, growths) Abnormal posture or movement

Head tilt Circling/spinning

Paralysis Behavior

Suddenly aggressive, quiet, loss of interest…

Laboratory Animal Care and Use Program and Facilities

Animal Observation

Condition of skin, Fur or Feathers Dirty, unkempt or ruffled fur

One of the most reliable signs of illness

Piloerection Alopecia Red, swollen or irritated skin Pruritus Bleeding

Laboratory Animal Care and Use Program and Facilities

Animal Observation

Mucus Membranes Pale

Anemia or lack of iron

Blue Cyanosis

Bleeding Swollen

Jaws and Teeth Malocclusion

Rodents and rabbits

Broken or missing

Laboratory Animal Care and Use Program and Facilities

Animal Observation

Perineal Area Vaginal discharge Prolapse

Rectum Vagina Uterus

Diarrhea Feces or blood staining

Constipation Inadequate feed or water intake

Laboratory Animal Care and Use Program and Facilities

Animal Observation

Tail Lesions

Trauma, infection, cancer

Sloughing Degloving

Lost tip Amputation, trauma

Feet Ulceration inflammation Lameness

Laboratory Animal Care and Use Program and Facilities

Animal Observation

Respiration Dyspnea Coughing Sneezing Rapid Shallow Nasal discharge

Laboratory Animal Care and Use Program and Facilities

Cause of Disease

Definition Any alteration of the normal anatomy of

physiology Malnutrition Infection Parasites Environment Contaminant

Laboratory Animal Care and Use Program and Facilities

Malnutrition

Water Fat Carbohydrates Proteins Vitamins Vitamin C

Minerals Experimental? Be aware of clinical signs expected

Laboratory Animal Care and Use Program and Facilities

Infection

Causes Bacteria Virus Fungi

Nonpathogenic Species specific Route Air Water Feed Feces

Laboratory Animal Care and Use Program and Facilities

Infection Submicroscopic, Microscopic, Macroscopic

Virus

Submicroscopic

Bacteria Microscopic

Fungus Microscopic (Macroscopic??)

Protozoa Microscopic

Helminth Macroscopic

Arthropod Macroscopic

Laboratory Animal Care and Use Program and Facilities

Parasites

Definition Organism that lives on or in an animal and

draws nourishment from the host Protozoa Single cell

Helminthe Worms

Arthropods Lice, ticks, fleas

Subclinical Altered data

Laboratory Animal Care and Use Program and Facilities

Environment

Temperature Noise Humidity Overcrowding Lighting Ventilation Vibration Avoid loud noises and music

Laboratory Animal Care and Use Program and Facilities

Contaminants

Air Water Bedding Cedar (↑ liver enzymes) ONLY

Hardwood, paper or corncob

Mold! Other Causes… Degenerative changes (aging, cancer) Allergy

Laboratory Animal Care and Use Program and Facilities

Transmission

Vector Organism that transmits a pathogen but does

not cause disease itself Fomite Nonliving items or materials that transmit an

infectious organism Direct contact Indirect contact (stable in the environment)

Laboratory Animal Care and Use Program and Facilities

Transmission

Aerosol Fecal-oral Mucous membranes Skin (puncture or wounds) High Quality Husbandry!!

SOPs PPE Designated facility workflow Sanitation (What water

temperature?) 180°F (82°C)

Laboratory Animal Care and Use Program and Facilities

Transmission

Sterile food HEPA filters Autoclave Room air pressure Positive or Negative?? Surgery Quarantine Biohazard

Laboratory Animal Care and Use Program and Facilities

Sentinel Animals

To monitor the health status in each colony animal room, isolator, and quarantine group.

Indicated by cage card Rotated on a schedule Necropsied or test (q 3 months) Direct Indirect Time Expense

Laboratory Animal Care and Use Program and Facilities

Sentinel Animals

Placed in a specific location on the rack

On schedule According to the SOP May miss some pathogens Mites Aerosol

Large animals Tested individually according

to signs of disease

Laboratory Animal Care and Use Program and Facilities

Rodent Health Surveillance Viral and Bacterial Pathogens RAT Sendai PVM SDAV KRV H-1 RPV RMV NS-1 REO GDVII

MOUSE EDIM ECTRO MAV1&2 MCMV K POLY Hantan M. pulmonis E. cuniculus Carbacillus MTLV

MOUSE Sendai PVM MHV MVM MPV1 MPV2 NS-1 GDVII REO LCMV Pasturella Pseudomonas H. hepaticus H. bilis

RAT LCMV MAV1&2 Hantan M. pulmonis E. cuniculus Carbacillus

Laboratory Animal Care and Use Program and Facilities

OSU Rodent Health Monitoring

Room Entry Housing requirements Disease Status

1 sterile, ventilated micro-isolator

cages, hood, sterile supplies Negative for viral & bacterial agents,

parasitology and Helicobacter

2 sterile, ventilated micro-isolator

cages, hood, sterile supplies

Negative for viral & bacterial agents, parasitology; Helicobacter positive

3 barrier ventilated micro-isolator or

static cages, hood Negative for viral agents and

Helicobacter

4 barrier ventilated micro-isolator or

static cages, hood Negative for viral agents, unknown

Helicobacter status

5 conventional housing &

techniques Negative for viral agents, unknown

Helicobacter status

6 Housing varies as listed in room Positive for viral, bacterial agent or

parasitology

Laboratory Animal Care and Use Program and Facilities

Diagnosis

Definition Investigation of the nature and cause of

disease History PE Testing Culture (urine, feces, wound) Evaluate blood cells Chemicals in the blood

Serum chemistry

Laboratory Animal Care and Use Program and Facilities

Detecting Parasites

Internal Cellophane tape test Fecal float

External Cellophane tape test Fur pluck (most common test) Direct exam (fleas, lice)

Regular testing

Laboratory Animal Care and Use Program and Facilities

Vaccinations

Large animals Ferrets

Canine distemper

Cats? Dogs? Long term Research requirements

Requires consultation with investigators

Laboratory Animal Care and Use Program and Facilities

Tuberculosis testing in NHP

Regular screening Highly contagious Where does it come

from? Human exposure

Rapid spread Personnel at risk Serious, rapid

progressive illness in NHP

Laboratory Animal Care and Use Program and Facilities

Tuberculosis testing in NHP

Tuberculin Noninfectious Purified protein

Frequency? Initial: within 7 days Twice more at 2 week intervals (total 3 in

quarantine) Every 6 months

Staff

Laboratory Animal Care and Use Program and Facilities

Tuberculosis testing in NHP

Eyelid Read? 48 or 72 hours

Six possible readings Isolation (quarantine) Animals suspected or known to be carrying

infectious disease Protect the colony from disease transmission Negative pressure air supply Husbandry last

Laboratory Animal Care and Use Program and Facilities

Tuberculin Test Reaction Grades Intradermal intrapalpebral test

0 No reaction observed 1 Bruise; extravasation of blood in

eyelid from injection 2 Varying degrees of redness

without swelling 3 Varying degrees of redness with

minimal or slight swelling 4 Obvious swelling with drooping of

lid 5 Swelling and/or necrosis with

eyelid closed

Laboratory Animal Care and Use Program and Facilities

What Grade?

Laboratory Animal Care and Use Program and Facilities

AALAT Chapter 14 Euthanasia

Laboratory Animal Care and Use Program and Facilities

AVMA Guidelines

Euthanatos “Good death” Ending animal life with minimal pain and

distress Rapid loss of consciousness

Cardiac arrest Respiratory arrest Loss of brain function

Responsibility of the institution to train personnel Humane proficient

Laboratory Animal Care and Use Program and Facilities

Methods

Categorized by species Acceptable Used alone consistently humane

Conditionally acceptable Require certain conditions to be met (weight,

specialized equipment) Unacceptable Inhumane Should not be used

Laboratory Animal Care and Use Program and Facilities

Animal Behavior

Avoid Distress Fear Anxiety

Gentle restraint Careful handling Sedation

Decreased blood pressure Delay onset of death by injectable

Laboratory Animal Care and Use Program and Facilities

Human Behavior

Attachment to the animals Human-research animal

bond Positive animal quality of life

Euthanasia related stress to care takers Psychological depression Alienation Job dissatisfaction Careless/callous animal

handling

Laboratory Animal Care and Use Program and Facilities

Human Behavior

Mechanisms for coping Training in grief coping skills Rotation of duties Peer support Professional support Education Time off

Laboratory Animal Care and Use Program and Facilities

Methods

Inhaled Agents Purified gas Slow exposure (gentle death takes longer) No leaks Neonatal animals may be resistant Minimize noise Groups

Same species Compatible Not crowded

Diving birds and mammals, reptiles, amphibians take longer

Laboratory Animal Care and Use Program and Facilities

Inhaled Agents

Anesthetics ACCEPTABLE WITH CONDITIONS for small

animals (<7 kg) Order of preference

Isoflurane Halothane Sevoflurane Enflurane Methoxyflurane Desflurane

Nitrous oxide never alone

Laboratory Animal Care and Use Program and Facilities

Inhaled Agents

Anesthetic delivery Open drop Face mask Chamber Vaporizer Vapors only (liquid is

irritating) Personnel exposure

Laboratory Animal Care and Use Program and Facilities

Inhaled Agents

CO Compressed gas cylinder with regulator Minimally distressful Aversive to rodents Hazardous to personnel!!

Training

Explosive (electrical outlets explosion proof) Chamber

High quality Well ventilated

Monitors present

Laboratory Animal Care and Use Program and Facilities

Inhaled Agents

N2 and Ar Compressed gas Minimal hazard to personnel Distressing to some species Revive with O2

CONDITIONS Some species O2 < 2% Regulated flow Heavily sedated animals

Laboratory Animal Care and Use Program and Facilities

Inhaled Agents

CO2 Compressed gas (NOT dry ice) Regulated flow

Pressure reducing regulator Displace 10-30% chamber volume per min

Maintain flow for 1 min after “death” Heavy, animals may climb above Prefilling is UNACCEPTABLE Must confirm

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Routes of Administration Immersion Topical Injection (Speed, fastest to slowest)

GENTLE IC IV IP IM SQ

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Barbituric Acid Derivatives ACCEPTABLE Depress the CNS Cardiac arrest Fast Regulated DEA Careful with carcass disposal

Pentobarbital Combinations ACCEPTABLE Combined with local anesthetic Neuromuscular agents NOT acceptable

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Tributame ACCEPTABLE for dogs Embutramide, chloroquine phosphate and

lidocaine IV only Not available in all countries

T-61 ACCEPTABLE Not available in the US Nonbarbiturate, nonnarcotic, curariform

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Dissociative Agents/Alpha-2 Adrenergic Receptor Agonists ACCEPTABLE WITH CONDITIONS

Approved euthanasia drugs unavailable As a secondary method

KCL and MgSO4 ACCEPTABLE WITH CONDITIONS

Unconscious or under general anesthesia IV or IC

Adjunctive method of euthanasia

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Chloral Hydrate/Alpha Chloralose NOT ACCEPTABLE

Aesthetically objectionable Previously acceptable for horses, ruminants and

swine

Alcohols ACCEPTABLE WITH CONDITIONS

Avertin (tribromoethanol) Prepared and stored appropriately

Tricaine Methanesulfonate ACCEPTABLE

Fish, some amphibians, reptiles MUST be buffered Protect from light, stored 4° or -20°F, remade monthly

Laboratory Animal Care and Use Program and Facilities

Noninhaled Agents

Benzocaine Hydrocloride ACCEPTABLE

Amphibians and fish

Agents used for fish ACCEPTABLE WITH CONDITIONS

Clove oil, isoeugenol, eugenol, 2-phenoxyethanol, quinaldine sulfate, metomidate and sodium hypochlorite

UNACCEPTABLE Strychnine Nicotine Caffeine Cleaning agents Solvents Pesticides Disinfectants

Laboratory Animal Care and Use Program and Facilities

Physical Methods

Penetrating Captive Bolt ACCEPTABLE WITH CONDITIONS

Placement and restraint important

Nonpenetrating Captive Bolt ACCEPTABLE WITH CONDITIONS

Suckling pigs, neonatal ruminants, turkeys

Stun

Blunt Force Trauma ACCEPTABLE WITH CONDITIONS Neonatal animals

Gunshot ACCEPTABLE WITH CONDITIONS Risky!

Laboratory Animal Care and Use Program and Facilities

Physical Methods

Cervical Dislocation ACCEPTABLE WITH CONDITIONS

Mice, rats <200g, rabbits and some birds

Decapitation ACCEPTABLE WITH CONDITIONS

Rodents, small rabbits, birds, fish, amphibians and reptiles SHARP!

Electrocution ACCEPTABLE WITH CONDITIONS

Cattle, sheep, swine and poultry NOT acceptable to tail, head, foot OR head to wet metal plate

Kill Traps ACCEPTABLE WITH CONDITIONS

Wildlife (when other methods have failed) Checked daily Approved by the International Org for Standards

Laboratory Animal Care and Use Program and Facilities

Physical Methods

Maceration ACCEPTABLE WITH CONDITIONS

Day old poultry and embryonated eggs

Focused Beam Microwave Irradiation ACCEPTABLE WITH CONDITIONS

Fix brain metabolites Small laboratory rodents

Thoracic Compression ACCEPTABLE WITH CONDITIONS

Wildlife biologist Small birds and mammals that are anesthetized

Adjunctive methods Applied when animals are anesthetized or unconscious

Decapitation Bilateral thoracotomy Exsanguination

Laboratory Animal Care and Use Program and Facilities

Physical Methods

UNACCEPTABLE Air embolism Burning Decompression Drowning Exsanguination Hypothermia Rapid freezing Smothering Thoracic compression was previously approved

as conditionally acceptable for free range birds

Laboratory Animal Care and Use Program and Facilities

Laboratory Animal Euthanasia

Research methods must be considered Approved by the IACUC Familiar with the species Well trained (for ANY method) Consider aesthetics Operator safety Economy

Laboratory Animal Care and Use Program and Facilities

Disposal

According to state and federal regulations Pentobarb and barbiturates are poisonous REMEMBER death must be confirmed! Lack of pulse Breathing Corneal reflex Toe pinch Mucous membranes Rigor mortis

Laboratory Animal Care and Use Program and Facilities

Questions??