LATG Chap. 4

1
LATG Chapters 4 5
Quality Assurance Occupational Health and Safety
2
Chapter 4 Quality Assurance

• ongoing verification system for the various
• aspects of laboratory operation
• - provides confidence
• - ensures reliability

3
Good Laboratory Practice
FDA regulations - must have a quality assurance
unit - responsible for monitoring studies -
members separate from personnel engaged in
study

4
Good Laboratory Practice

• Requirements
• Maintain a copy of master schedule indexed by
  test article
• Maintain copies of all protocols
• Inspect studies periodically
• No deviations from approved protocol or SOP
• Review final report for conformity
• List available to FDA of methods of recording and
  indexing

5
Good Laboratory Practice

• Standard Operating Procedures ( SOPs)
• Handling of test and control articles
• Maintenance and calibration of equipment
• Animal care
• Animal room preparation
• Test system observations

6
SOPs

• Laboratory tests
• Handling of dead or moribund animals
• Necropsy of animals
• Collection and identification of specimens
• Histopathology
• Data handling, storage, and retrieval
• - Transfer, placement, and identification of
  animals

7
Good Laboratory Practice

• Equipment Design
• of appropriate design and capacity function
  according to
• protocol located suitably
• Maintenance and Calibration
• written SOPs to cover equipment maintenance
• actions to be taken for malfunction
• designate personnel responsible for each
  operation
• written records of maintenance to include dates
  and any revisions

8
GLP

• Animal Care
• SOPs for feeding, handling, housing, and care
• facilities for quarantine, health
  evaluations
• ID, separate rooms for species
• - analysis of feed and H2O

9
Good Laboratory Practice

• Protocol
• each study must have an approved written
  protocol that indicates study objectives and all
  methods to be used
• - all changes and reasons for changes must be
  documented and signed by the study director
• Nonclinical Study
• - all data recorded directly, promptly, in ink
• - entries dated signed changes not to obscure
  original

10
Components of a Good Quality Assurance Program

• Procuring Animals
• Animal Husbandry
• Animal Health
• Genetic Monitoring
• Equipment Monitoring
• Monitoring Climate
• Microbial Monitoring of Environments
• Monitoring Techniques

11
Components
Procuring Animals - specifications according to
needs - suppliers in compliance with
regulations Animal Husbandry - Feed (expiration
dates autoclavable certified) - Water Quality
( automatic devices RO contaminant
analysis) - Bedding (processed bedding best
lookout for dust foreign materials,
feces) - Disposal Services (timely and thorough
use discretion)
12
Components
Animal Health Microbial monitoring of live
animals - 10 14 days to develop immune
response - serology - sample size Loge
(1.0 C) C confidence level Loge
(1.0 M) M morbidity Sentinel
Animals - should resemble study animals -
immunocompetent - indirect association (soiled
bedding airborne transmission) - in place 3 6
weeks
13
Components

• Microbial Monitoring of Environments
• - agar plates incubated for 24 hours at 37ºC
  (CFUs)
• frequency tailored to need (mouse cage vs. dog
  run)
• Equipment Monitoring
• - autoclaves (autoclave tape chemically treated
  paper)
• - Bacillus stearothermophilus spores
• - cage washers (adhesive plastic tape changes
  from
• silver to black at 82 /- 1ºC at 13 secs)

14
Components

• - water bottles filling stations automatic
  watering system
• - (test for Salmonella Shigella Pseudomonas
  coliforms)
• ventilated racks/hoods
• (filter replacement airflow hoods certified)

15
Components
Monitoring Climate Ventilation within
microenvironment - elevated temp humidity
ammonia increases
susceptibility to infectious agents Noise
adverse effects gt 85 dB - house noisy animals
away from rodents and rabbits - hearing
protection for humans - reduces fertility
audiogenic seizures - loud equipment away from
animal rooms
16
Components
Illumination gt 75 foot-candles retinal damage
to albino animals - 30 foot-candles at one meter
above floor adequate - 12/12 light cycle
(changes detrimental to breeding)
17
Components

• Genetic Monitoring (inbred strains)
• 1. It must be accurate and precise and therefore
  highly
• reproducible
• 2. It must be relatively easy to carry out
• 3. It must be efficient
• 4. It must be economical
• done by qualified experienced technicians

18
Components

• Monitoring Techniques
• Biochemical Markers enzymes in animal tissue
  detected using electrophoresis and histochemical
  staining
• Immune Markers antigens found on cell
  surfaces erythrocyte histocompatibility
  antigens common
• Mandibular Morphometrics measures size and
  shape
• Breeding Performance sudden increase in first
  litter size
• Breeding Index
• Lane Petters Q Index
• Polymerase Chain Reaction ( PCR) newest
  method utilizes DNA patterns

19
OccupationalHealth and Safety
Chapter 5
20
ILAR
1997 - Occupational Health and Safety in the Care
and Use Of Research Animals Considered the
standard by which all occupational health
programs involving research animals are measured.

21
Risk Assessment
Knowing what hazards are present, then
determining the level of protection necessary to
minimize danger Physical hazards (equipment,
bites, sharps) Chemical and biological
(carcinogens infectious agents) Allergens Zoonose
s
TRAINING !!
22
Physical and Chemical Hazards
1. Animal bites, scratches, kicks, related
injuries - know your species proper handling 2.
Sharps improper disposal 3. Flammable materials
4. Pressure vessels cylinders chained
autoclaves 5. Lighting 6. Electricity report
defects
23
Physical and Chemical Hazards

• UV Radiation eye skin protection
• 8. Lasers protection from beams
• 9. Ionizing Radiation - type, dose, half-life
• - proper shielding, monitoring exposure, waste
  handling
• 10. Housekeeping safe work environment
• 11. Ergonomic Hazards- lifting repetitive
  motion
• 12. Machinery good maintenance

24
Physical and Chemical Hazards
13. Noise hearing protection 14. Chemicals
nature quantity mode and duration of exposure
MSDS fume hoods 15. Infectious Agents and DNA
virulence, pathogenicity, communicability, and
route of spread
25
Allergens
Laboratory animal allergy (LAA) - MOST COMMON
occupational health problem - dander, saliva
urine proteins Prevention - face mask -
gloves/ washing hands - surgical gowns/ lab
coats - filtered cage tops


26
Allergens
- positive pressure air flow - HEPA filtered
exhaust systems - frequent air changes -
biological safety cabinets
27
Zoonoses
Diseases of animals transmissible to humans TB
testing NHPs Toxoplasmosis pregnant
women Immunizations Tetanus Rabies random
source dogs/cats bats Vaccinia Hepatitis B
serum, blood, tissues from humans/apes
28
Personal Protection
Protective apparel lab coats, safety goggles,
face shields, gloves Special equipment -
biological safety cabinets
29
Personal Protection
Respirators - toxic chemical vapors gases -
BSL 3 4 agents - allergies fit test for
proper mask-to-face seal OSHA specifications
30
Personal Protection
Eye and Skin Protection - Safety glasses -
Gloves primary barrier (latex best)
31
Personal Protection
Biological Safety Cabinets Class I and II - air
curtain creates barrier Class II - laminar flow
or biosafety hood Class III- physical barrier
prevents direct contact HEPA filters remove
99.97 of particulates 0.3 microns or
larger Fume hood for gaseous constituents
32
Waste Disposal
Must comply with institutional, local, state and
federal regulations General waste soiled
bedding, carcasses from healthy
animals Hazardous waste toxic chemicals,
infectious and radioactive materials,
contaminated or diseased animal carcasses -
separated from other waste - clearly identified
(universal symbols) - use protective gloves and
clothing
33
Waste Disposal
- packaged in leak resistant paper/cardboard,
stainless steel, polymers - rigid , puncture
resistant sealable containers for sharps - wet
waste double bagged
34
Waste Disposal
Infectious waste steam autoclaving best for
decontaminating prior to removal from
biocontainment area Incineration best for
treating large volumes of infectious
waste Radioactive waste separated labeled
according to isotope and form Mixed waste
use caution for treatment
35
Emergencies and First Aid
Emergency plan to cover both humans and animals
- evacuation routes - meeting places -
facilities for medical treatment - procedures
for reporting emergencies Assist injured
Provide first aid Use resources (fire dept
medical help) Plan to evacuate and save animals