Transcript January 2016 – Biosafety – Working Safely in a BSL-3

BIOSAFETY- WORKING
SAFELY IN A BSL-3
LABORATORY
Parampal Deol, Ph.D, MBA, Director R&D Microbiology
Anne Hutchins, Ph.D, Associate Staff Scientist
Agenda
•Biosafety: Principles/concepts
•Areas of Biosafety
•Biological Risk Groups
•Standards
•Types of Biosafety Laboratories
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Engineering controls
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Best practices
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Operational Controls
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Administrative controls
•BSL 3 laboratory in bMX
•Select Agents
Biological Safety
•Biosafety - the discipline addressing the safe handling &
containment of infectious microorganisms & hazardous
biological materials
• application of knowledge & the use of appropriate techniques & equipment
• prevent personal, laboratory & environmental exposure to potentially
infectious agents or biohazards
Source: BMBL, 5th Ed.
Areas of Biosafety
• Laboratory Safety
• Respiratory Protection (TB)
• Bloodborne pathogens (BBP) &
• Bioterrorism & select
other infectious agents
agents/toxins
• Recombinant DNA (rDNA)
• Mold & indoor air quality
• Biological waste disposal
• Occupational safety & health in
• Infectious substance shipping
the use of research animals
Hazard Classifications of Microbial Agents
Classification is based on Human Disease
•Risk Group 1: Agents not associated with disease
• K12 strain of E. coli
•Risk Group 2: Agents associated with disease
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rarely serious & preventive / therapeutic interventions are often
available
• Salmonella species, HIV, Listeria monocytogenes
•Risk Group 3: Agents associated with serious or lethal disease
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preventive/ therapeutic interventions may be available
• Mycobacterium tuberculosis, Eastern Equine Encephalitis (EEE) Virus, Bacillus anthracis
•Risk Group 4: Agents likely to cause serious or lethal disease
• preventive or therapeutic interventions are not usually available
• Ebola virus, Lassa virus, Variola virus (smallpox)
Laboratory Design and Operational Practices
• Biosafety in Microbiological and Biomedical Laboratories
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(BMBL) 5th edition
Centers for Disease Control and Prevention (CDC)
National Institute of Health (NIH)
Federal Select Agent Program (FSAP)
World Health Organization (WHO)
United States Department of Agriculture (USDA)
Regulations & Guidelines
• Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, CDC/NIH (BMBL)
• NIH Guidelines for Research Involving Recombinant DNA Molecules, April 2002 as
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amended
Occupational Safety and Health Administration (OSHA) Bloodborne Pathogens Standard
29 CFR 1910.1030
Respiratory Protection including Tuberculosis 29 CFR 1910.134 (independent standard
1910.139 repealed)
State regulations concerning biological waste disposal
IATA Dangerous Goods Regulations, DOT 49 CFR 171-185, & USPS 39 CFR Part 111
Select Agents and Toxins Regulations (HHS/CDC 42 CFR 73; USDA/APHIS 7 CFR 331 &
9 CFR 121)
Biosafety: Where to Start?
Your laboratory wants to start working with an infectious
agent and you need to have a lab space or renovate
current lab.
How do you decide what changes need to be made to the
current space to safely handle the bacteria.
What are the guidelines?
Biosafety Measures Illustrated
Microbiological
Practices
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Standard
Special
Administrative Controls
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Training
Operations
Maintenance
Risk Assessment
Emergency Response
Plans
Safety Equipment
(Primary Barriers)
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PPE
BSCs
Facility Design
(Secondary Barriers)
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Lab Inspections
Biosecurity
Biosafety Manual
SOPs
Medical Surveillance
Respiratory Protection
Inventory
Record Keeping
Principles of Biosafety
Containment
• microbiological practices
• safety equipment
• facility safeguards
• protect laboratory workers, the environment & the public from
exposure to infectious agents that are handled & stored in the
laboratory
Risk assessment
• process
• enables the appropriate selection of microbiological practices,
safety equipment & facility safeguards that can prevent laboratoryassociated infection
The backbone of the practice of biosafety is risk assessment
WHO Biosafety Manual, 3rd ed.
Biological Containment
• Laboratory Practice & Technique
• Adherence to standard microbiological practices & techniques
• Awareness of hazards & training
• Biosafety or laboratory Standard Operating Procedures (SOPs)
• Safety Equipment (Primary Barriers)
• Biological safety cabinets (BSC)
• Safety centrifuge cups/sealed rotor heads
• Personal protective equipment (PPE)
• lab coats, gowns, gloves, safety glasses, face shields, shoe covers,
respirators, Tyvek suit
• Facility Design & Construction (Secondary Barriers)
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Restricted Access from public
Autoclave
Directional airflow
HEPA filter units
Airlocks as lab entrances
Biological Containment
Biological Safety Cabinets (BSC)
Class I, Class II & Class III
Biological Safety Level - 1 (BSL-1)
• Standard microbiological
practices:
• Hand washing sink
• No eating, drinking, smoking,
applying cosmetics, handling
contact lenses, or storing food
• No mouth pipetting
• Safe handling of sharps
• Decontaminate work surfaces
after completion of work, & any
spill or splash; any waste
generated
• Biohazard symbol when
infectious agents present
• Wear lab coats or gowns,
protective eye wear, gloves
• Windows to exterior have
screens
WHO Biosafety Manual
3rd Ed., 2004
Biological Safety Level – 2 (BSL-2)
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All of BSL-1 practices plus:
Access is restricted to trained
personnel
Lab doors should be selfclosing & have locks
BSC used when potential for
aerosol production
Sealed centrifuge cups/rotors
Eyewash station must be
readily available
Method for decontaminating
laboratory waste should be
available in the facility
WHO Biosafety Manual
3rd Ed., 2004
Biological Safety Level – 3 (BSL-3)
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All of BSL-2 practices plus:
 Lab is separated from
unrestricted traffic flow
 Access to lab is restricted to
entry through a series of 2 selfclosing doors (ante-room)
 All manipulations must be
conducted in a BSC, other
containment, or when wearing
proper PPE
 Protective clothing with a solid
front (tie-back or wrap-around
gowns, scrubs, coveralls) is
worn
 Eye, face, & respiratory
protection may be worn
 Hands-free sink near exit door
WHO Biosafety Manual
3rd Ed., 2004
Biological Safety Level – 4 (BSL-4)
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All of BSL-3 practices plus:
Generally a separate facility;
dedicated mechanical, electrical,
plumbing, HVAC
Agents handled in Class III BSC or
with one-piece positive pressure
personnel suit
Shower out practices
Lab clothing decontaminated prior to
laundering
Liquid effluent decontaminated prior
to release to sewer
Redundant supply fans
recommended, redundant exhaust
fans required, interlocked supply &
exhaust
CDC workers in BSL-4
www.myajc.com
Biological Safety Level - 3 Laboratory Design
• Containment, ability to clean
• Full containment
• Controlled access
• Surfaces design for cleaning
• Sustained directional (negative) airflow
• Visual monitoring device at entry, gown in and prior to entrance in lab space
• Utilities
• Lab air supply and exhaust fans with HEPA
• HEPA filters certified and replaced as needed
• Utilities (exhaust) are separated
• Decontamination/Spill Management
• Waste disposal autoclave within lab
• Equipment (surface)
• Spill cleaning
BSL-3 in at bioMerieux in Durham
• Emergency Response
• Natural Disasters
• Tornado warnings
• Transport
• Import/export of cultures
• Operational SOPs/GMP
• All records electronic, if possible
• Medical Surveillance
• Sustainability and permits
• Annual certification
Select Agent Regulations
What are the Select Agents?
•Biological agents /toxins pose a severe threat to human
health
•65 agents and toxins
• Reconstructed 1918 Influenza virus
• Ricin toxin
• Ebola virus *
• Bacillus anthracis*
• Francisella tularensis*
* denotes Tier 1
• Established to keep the agents/toxins out of possession of
individuals who may misuse them
•USA Patriot Act
•Public Health Security and Bioterrorism Preparedness and
Response Act of 2002
•The Select Agent Regulations (7 CFR Part 331, 9 CFR Part
121, and 42 CFR Part 73) www.selectagents.gov
2001 Anthrax Letter
22 illnesses
5 deaths
Cost $23 million
www.selectagent.gov
Who Can Possess Select Agents?
Individuals
•FBI security clearance
Entities
•Facility Design Requirements
•Biosafety Plan
•Security Plan
• Physical, control of select agents, inventory, and information system
•Incident Response Plan
• Natural Disaster events
• Failures- spills, equipment failures, unsecured containment, loss of power
Need to Train and SA requires annual drills
Tier 1 Select Agents
Greatest risk of deliberate misuse for mass casualties or devastating effect to the
economy, critical infrastructure, or public confidence
Requires additional measures such as:
• personnel suitability assessment
• occupational health program
•visitor procedures
• response time of security forces within 15 minutes
• incident response procedures
• annual training on reporting suspicious behavior
• > 3 physical security barriers
• intrusion detection system (IDS) with backup during power failure
Non-Tier 1 (Clinical laboratories)
• Can still test for and identify a
Tier 1 agent
• Can not ‘possess’ a Tier 1
agent
• Have 7 days to transfer or
destroy a Tier 1 agent
• State Public Health Laboratories
• Laboratory Response Network
(LRN)
LRN Network
(www.cdc.gov)