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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??