Quality AssuranceOccupational Health and Safety

LATG Chapters 4 & 5

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

*

OccupationalHealth and Safety

Chapter 5

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

Eye and Skin Protection- Safety glasses- Gloves – primary barrier (latex best)

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

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 aidUse resources (fire dept; medical help) Plan to evacuate and save animals