Biological Safety Training
East Carolina University
Office of Prospective Health/Biological Safety
Introduction to Biological Safety
Biological Hazards are divided into 4 Biosafety
Levels
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–
–
–
BSL 1
BSL 2
BSL 3
BSL4
Biosafety levels define the lab requirements, protective
clothing, and work practices
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Definitions
BSL 1 (BMBL)
– Not known to cause disease in healthy human
adults
– Pose minimal hazards under ordinary
conditions of handling
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 1
• Standard Practices
– Use Mechanical Pipetting devices
– No Eating, Drinking, Smoking in
Lab
– Minimize splashes and aerosols
– Decontaminate work surfaces
– Safe handling of sharps
– Wash Hands before leaving lab
– Biohazard sign is posted
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 1
• Protective Clothing
– Lab Coat
– Gloves
– Eye Protection
(if splashes are likely)
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Definitions
• BSL 2 (BMBL)
– Work with well-characterized agents not known
to cause disease in healthy adult humans;
minimal hazard to laboratory personnel and the
environment
Examples:
Salmonella,
Hepatitis B Virus
Measles Virus (not aerosolized)
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Definitions
• Other Materials Handled at BSL 2
(Requires compliance with OSHA BBP Standard)
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–
–
–
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Human Blood
Human Tissues
Human Cell Lines
Biotoxins
Viral Vectors
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 2
• Standard Practices
– All requirements for BSL 1 plus:
• Access to laboratory is limited or restricted when
work is being conducted; door is closed
• Personnel have specific training in handling
pathogenic agents
• Extreme precautions are taken with contaminated
sharp items
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 2
• Standard Practices cont.
•
•
•
•
Written Biosafety Procedures
Report Spills
Minimize aerosol generation
Personnel receive immunizations or testing (e.g.
Hep B vaccine or TB skin Testing)
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 2
•
Personal Protective Equipment
– Lab Coats
– Gloves
• Double Gloving when appropriate
– Eye Protection - Safety Glasses/Shield
– Biosafety Cabinets or sealed safety centrifuge
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 3
• BSL 3 (BMBL)
– Use of indigenous or exotic agents which may
cause serious or potentially lethal disease from
exposure by the inhalation route.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 3
• Standard Practices
– All requirements for BSL 2 plus:
•
•
•
•
Very limited lab access
2 doors in a series to access lab
Able to decontaminate entire lab
Special exhaust ventilation
(Not re-circulated, no airflow outward)
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 3
• Standard Practices –cont.
– High level of training
– Personnel receive vaccinations if available
– Work in Biosafety cabinets and/or respirator
used
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 3
• Examples
– Mycobacterium Tuberculosis
– Hanta virus
– SARS
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 4
• BSL 4 (BMBL)
– dangerous and exotic agents with a high risk of
aerosol-transmitted laboratory infections and
life-threatening disease.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 4
• Requirements
– All requirements for BSL3 plus:
• Class III Biosafety cabinet
or positive pressure suits
• Shower/Change rooms
• Clothing Autoclaved before laundering
• Air Locks
• All waste contained and
decontaminated onsite
• Space Suits
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Space Suit
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Level 4
• Examples
– Ebola Virus
– Monkey B Virus
– Marburg Virus
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Safe Work Practices for all
Levels
• Wash hands after work; when removing
gloves; before leaving lab
• No eating, drinking, applying cosmetics,
handling contact lenses in the lab.
• Maintain labs in clean, orderly fashion.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Safe Work Practices for all
Levels
• Limit access to lab when work with
organisms is in progress
• Use good microbiological techniques
• No mouth pipetting
• When possible use plastic instead of glass
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Safety
• Sharps include needles, syringes, razor
blades, lancets, slides, scalpels, pipettes,
micropipettes, pipette tips, broken plastic or
glassware, and other devices capable of
cutting or piercing the skin.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Safety
• Contaminated needles are not bent,
recapped, or removed.
• If recapping is required, use a mechanical
device or a one handed technique.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Safety
• Safety devices or
alternatives to needles
should be used when
available.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Disposal
• Sharps containers for
disposal of these items
are conveniently
located and easily
accessible.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Disposal
• Syringes and syringes without a needle
attached go into a sharps container
• Contaminated micropipettes, pipette tips,
and Pasteur pipettes are discarded in a
puncture-resistant container or a sharps
container for disposal.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Sharps Disposal
• Don’t place needles or
sharps in soft-sided
bags.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Laminar Flow Equipment
BioSafety Cabinets (BSCs)
Protect personnel and the environment from
hazardous particulates and from infectious agents
Laminar Flow Clean Benches (LFBs)
Non Hazardous work only
Protect work from contamination
*Do not protect personnel*
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
BioSafety Cabinets
• 3 Classes
• All exhaust is HEPA
filtered before leaving
the cabinet
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
NIH Guidelines for Use of
Recombinant DNA apply to all
work at ECU,
not just NIH-funded work.
The university can be sanctioned
for failure to comply.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
NIH Guidelines may apply to
work with:
•
•
•
•
•
•
Recombinant DNA or RNA
Genetic/Recombinant Techniques, molecules or organisms
Transgenic plants or animals or knock-out organisms
Viral or host vector systems or plasmids
Human gene transfer
Transfer of drug-resistance to microorganisms not known to
acquire it naturally
• Formation of DNA or RNA genes to synthesize toxin
molecules
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
At ECU, all work which may be
subject to the NIH Guidelines
must be reviewed by the
Biological Safety to determine
whether the work is “NIH
exempt.”
The researcher may not make this
determination independently
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Submission of a Biological Safety
registration to the Institutional Biological
Safety Committee will ensure that full
review per the NIH Guidelines occurs.
Form is available on the Prospective Health
website http://www.ecu.edu/csdhs/prospectivehealth/Biological-Safety-Office-ofProspective-Health.cfm
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Some work must be approved by NIH
as well as by the local Institutional
Biological Safety Committee:
• Transfer of Drug Resistance to
microorganisms not known to acquire
naturally
• Formation of genes to synthesize toxins
lethal to vertebrates at an LD50 of < 100
nanograms per kg body weight (e.g., tetanus
toxin)
• Human gene transfer
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Drug resistance is of concern if it could
compromise the use of the drug to control
disease in humans, veterinary medicine or
agriculture.
Only NIH may determine whether the drug
resistance can compromise therapeutic
use!
This is not a local or researcher decision.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Biological Safety interfaces with
the Human Subjects
Institutional Review Board
(IRB) and Institutional Animal
Care and Use Committee
(IACUC) to ensure that the NIH
Guidelines are met at ECU.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
The IBC must report significant
violations of the NIH
Guidelines, as well as researchrelated accidents and illnesses
to the NIH Office of
Biotechnology Affairs.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
Contact Biological Safety, Office
of Prospective Health for
questions.
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm
For more information see the
ECU Biological Safety Manual
Office of Prospective Health/Biological Safety
Biological Safety Committee
East Carolina University
http://www.ecu.edu/cs-dhs/prospectivehealth/customcf/biosafety/BIOLOGICAL_SAFETY_MANUAL%20_WEB.pdf
East Carolina University
Office of Prospective Health/Biological Safety
http://www.ecu.edu/cs-dhs/prospectivehealth/index.cfm