(CAREC) PAHO/WHO Serving 21 Member Countries in the English & Dutch - Speaking Caribbean An overview of Laboratory Safety Biosafet y Dr. Ashok Rattan, Medical Microbiologist & Laboratory Director, CAREC
May 07, 2015
(CAREC)PAHO/WHO
Serving21 Member Countriesin the English & Dutch -Speaking Caribbean
An overview of Laboratory Safety
Biosafety
Dr. Ashok Rattan,Medical Microbiologist &
Laboratory Director,CAREC
Caribbean Epidemiology Centre (CAREC) PAHO/WHO
Before entering the water to swim you must learn all you can about the sharks that may lurk there
SAFETY IN THEMYCOBACTERIOLOGYLABORATORY
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Protection:– workers– “products”– co-workers– lab support personnel– environment
IntroductionWhy Biosafety Practices?
2.1
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Safety practices should be applied throughout the testing process:
Pre- analytical– Specimen collection
– Specimen preparation
– Specimen transport
Analytical– Testing
Post-analytical– Disposal
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Examples of Laboratory Hazards
Growth of microorganisms
Water baths
Aerosols, splashing, tube breakage
Centrifuge
Accidental inoculation, aerosol, spillage
Needles
HazardOperation / Equipment
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Safety Resources
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Safety Resources
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IntroductionChain of Infection
Reservoir of pathogenReservoir of pathogen
Portal of escapePortal of escape
TransmissionTransmission
Route of entry/infectious doseRoute of entry/infectious dose
Susceptible hostSusceptible host
Incubation periodIncubation period
Risk A
ssessment
PPE
Immunization
Surveillance
Practices/
Equipment
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BSL1 - agents not known to cause disease. BSL2 - agents associated with human disease. BSL3 - indigenous/exotic agents with potential for aerosol
transmission; disease may have serious or lethal consequences. BSL4 - dangerous/exotic agents which pose high risk of life-
threatening disease.
Principles Biosafety Levels of agents
2.1
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Designing for Safety
General requirements– Facility design
– Water supply/sinks for hand washing
– Ventilation
– Standard lab practices
Safety equipment– Personal protective equipment (PPE)
– Biosafety cabinets
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Biosafety Level 1 Standard Microbiological Practices 1/3
Restrict or limit access when working
Prohibit eating, drinking and smoking in the laboratory
Pipetting by mouth strictly forbidden
2.3
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Biosafety Level 1 Standard Microbiological Practices 2/3
2.3
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Standard practices also include: 3/3
Keep work areas uncluttered and clean No food in lab refrigerator Minimize splashes and aerosols Decontaminate work surfaces daily Maintain insect & rodent control program
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Biosafety Level 2 Facility Design (Secondary Barriers) 1/8
Requirements: Laboratories have lockable doors Sink for hand washing Work surfaces easily cleaned Bench tops are impervious to water Sturdy furniture
2.4
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Biosafety Level 2Laboratory Facilities (Secondary Barriers) 2/8
BSL-1 Facilities PLUS:– Autoclave available– Eyewash station
available
2.4
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Requirements:– Location - separated from public areas
– Structure - normal construction
– Ventilation - directional
Biosafety Level 2 Facility Construction (Secondary Barrier) 3/8
2.4
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Biosafety Level 2Safety Equipment (Primary Barriers) 4/8
In addition to BSL-1: Use biosafety cabinets (class II) for work with
infectious agents involving:– Aerosols and splashes– Large volumes
– High concentrations
2.4
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Biosafety Level 2Special Practices 5/8
Needles & Sharps Precautions – Use sharps containers– DON’T break, bend, re-sheath or reuse
syringes or needles
2.4
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Biosafety Level 2Special Practices 6/8
Needles & Sharps Precautions DON’T place needles or sharps in office waste
containers
2.4
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Biosafety Level 2Special Practices 7/8
Needles and Sharps Precautions – DON’T touch broken glass with hands
2.4
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Identify “clean” and “contaminated” areas – Use appropriate warning signs
Decontaminate work surfaces Report spills and accidents Remove gloves, lab coats before leaving
work area No animals in laboratories
Biosafety Level 2Special Practices 8/8
2.4
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Biosafety Level 3Laboratory Facilities (Secondary Barriers) 1/5
BSL-1 and 2 Facilities PLUS:
– Separate building or isolated zone
– Double door entry
– Directional inward airflow
– Single-pass air; 10-12 air changes/hour
2.5
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Biosafety Level 3Laboratory Facilities (Secondary Barriers) 2/5
BSL-1 and 2 Facilities PLUS (cont.):
– Enclosures for aerosol generating equipment
– Room penetrations sealed
– Walls, floors and ceilings are water resistant for easy cleaning
2.5
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Biosafety Level 3Safety Equipment (Primary Barriers) 3/5
BSL-1 and 2 Safety Equipment PLUS:– BSC class
II or III to manipulate infectious material
2.5
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Biosafety Level 3Safety Equipment (Primary Barriers) 4/5
BSL-1 and 2 Safety Equipment PLUS:– Respiratory protection may be indicated
2.5
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BSL-2 Special Practices PLUS:– Work in certified BSC– Use bioaerosol-
containing equipment– Decontaminate spills
promptly
Biosafety Level 3Special Practices 5/5
2.5
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Biosafety Level 4 – Maximum Containment BSL -3 practices plus:
– Clothing change before entering laboratory– Shower on exit– All materials decontaminated on exit from facility
Safety Equipment:– Class III Biosafety cabinet– Class I or II biosafety cabinet– WITH full-body, air supplied,
• positive personnel suit
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Biological Waste
Types– cultures, stocks, isolates– materials containing or contaminated with blood– sharps– pipettes, wrappers, tips– All materials used in the lab
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Specimen Disposal
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DecontaminationDecontamination Sterilization Disinfection
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DecontaminationDefinition
SterilizationThe use of a physical or chemical procedure to
destroy all microbial life, including large numbers of highly resistant bacterial spores.
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DisinfectionThe use of a physical or chemical procedure to
virtually eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial endospores) on inanimate objects.
DecontaminationDefinition
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DecontaminationMethods
Heat Chemical Radiation
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Types– Moist – steam– Dry– Incineration
*The most effective method of sterilization
DecontaminationHeat
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Types– Liquids, i.e. chlorox,
hydrogen peroxide– Gases, i.e. ethylene oxide
DecontaminationChemical
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General Lab Use - Hypochlorite Solutions– Large Spills/Large Organic Load
• undiluted from bottle– Small Spills/Virus Inactivation
• 10% - 1:9– General Surface Disinfection
• 1% - 1:99
DecontaminationChemical
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In case of a spill
Wear disposable gloves Cover large blood spill with paper towels and
soak with 1% (10000 ppm) of household bleach and allow to stand for at least 5 minutes
Small spill - wipe with paper towel soaked in 1% bleach
Discard contaminated towels in infective waste containers
Wipe down the area with clean towels soaked in a same dilution of household bleach
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Safety Documentation & Records
Laboratory Safety Manual - Policies and Procedures Sample Contents:
– Housekeeping– Personal protection– Safe decontamination of equipment– Decontamination & Waste Disposal– Emergency procedures
In-lab first aid Accidental injury Post exposure prophylaxis Contacts
– Personnel responsibilities Hygiene
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Action Plan for Implementing Safety Practices Identify hazards Assess level of risks
– Prioritize risk Establish and implement safety polices and
procedures Conduct safety specific training
– Must be a priority– Communication is key
Perform regular audits and assessments
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In Case of Exposure
Be ready for the emergency before hand- Familiar with exposure specific
policies- Conduct drills - Keep post exposure medicines
available- Check periodically for stock and
expiry of medicines Report immediately Go to the nearest, first available doctor
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Post Exposure
Write a report and reasons for accident Actions taken to avoid future accidents Training
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SAFETY IN THEMYCOBACTERIOLOGYLABORATORY
Caribbean Epidemiology Centre (CAREC) PAHO/WHO