Chapter 4 – Quality Assurance
• ongoing verification system for the various aspects of laboratory operation
- provides confidence
- ensures reliability
Good Laboratory Practice
FDA regulations - must have a quality assurance unit
- responsible for monitoring studies
- members separate from personnel engaged in study
Requirements
1. Maintain a copy of master schedule indexed by test article
2. Maintain copies of all protocols
3. Inspect studies periodically
4. No deviations from approved protocol or SOP
5. Review final report for conformity
6. List available to FDA of methods of recording and indexing
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
Good Laboratory Practice
-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
SOPs
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
Good Laboratory Practice
Animal Care
- SOPs for feeding, handling, housing, and care
- facilities for quarantine, health evaluations
- ID, separate rooms for species
- analysis of feed and H2O
GLP
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
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
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)
Components
Animal HealthMicrobial 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
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)
*
- water bottles; filling stations; automatic watering system
- (test for Salmonella; Shigella; Pseudomonas; coliforms)
-ventilated racks/hoods
- (filter replacement; airflow; hoods certified)
Components
Components
Monitoring Climate
Ventilation: within microenvironment - elevated temp; humidity; ammonia increases susceptibility to infectious agents
Noise: adverse effects > 85 dB- house noisy animals away from rodents and rabbits- hearing protection for humans- reduces fertility; audiogenic seizures- loud equipment away from animal rooms
Illumination: > 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)
Components
Components
Genetic Monitoring (inbred strains)1. It must be accurate and precise and therefore highly
reproducible2. It must be relatively easy to carry out3. It must be efficient4. It must be economical
-done by qualified experienced technicians
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 IndexLane Petters Q Index
• Polymerase Chain Reaction ( PCR) – newest method utilizes DNA patterns
*
ILAR
1997 - Occupational Health and Safety in the Care and Use Of Research Animals
Considered the standard by which all occupational health programs involving researchanimals are measured.
Risk Assessment
Knowing what hazards are present, then determiningthe level of protection necessary to minimize danger
Physical hazards (equipment, bites, sharps)Chemical and biological (carcinogens; infectious
agents)Allergens
Zoonoses
TRAINING !!
Physical and Chemical Hazards
1. Animal bites, scratches, kicks, related injuries
- know your species; proper handling2. Sharps – improper disposal3. Flammable materials 4. Pressure vessels – cylinders chained;
autoclaves5. Lighting6. Electricity – report defects
Physical and Chemical Hazards
7. UV Radiation – eye & skin protection8. Lasers – protection from beams
9. Ionizing Radiation - type, dose, half-life- proper shielding, monitoring exposure, waste
handling10. Housekeeping – safe work environment11. Ergonomic Hazards- lifting; repetitive
motion12. Machinery – good maintenance
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
Physical and Chemical Hazards
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
*
- positive pressure air flow
- HEPA filtered exhaust systems
- frequent air changes
- biological safety cabinets
Allergens
Zoonoses
Diseases of animals transmissible to humansTB testing – NHPsToxoplasmosis – pregnant women
ImmunizationsTetanusRabies – random source dogs/cats; batsVacciniaHepatitis B – serum, blood, tissues from
humans/apes
Personal Protection
Protective apparel – lab coats, safety goggles, face shields, gloves
Special equipment - biological safety cabinets
Personal Protection
Respirators- toxic chemical vapors; gases- BSL 3 & 4 agents- allergies
“fit test” for proper mask-to-face seal
OSHA specifications
Personal Protection
Biological Safety CabinetsClass I and II - air curtain creates barrierClass II - laminar flow or biosafety hoodClass III- physical barrier prevents direct contact
HEPA filters – * remove 99.97% of particulates 0.3 microns or larger
Fume hood – for gaseous constituents
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 radioactivematerials, contaminated or diseased animal carcasses
- separated from other waste- clearly identified (universal symbols)- use protective gloves and clothing
- packaged in leak resistant paper/cardboard, stainless steel, polymers
- rigid , puncture resistant sealable containers for sharps
- wet waste double bagged
Waste Disposal
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