RECOGNIZING
THE BIOSAFETY LEVELS
Biosafety Levels
AUTHORIZED PERSONNEL ONLY
Prepared by :
Debbra Marcel, Monoclonal Antibody Section,
Veterinary Research Institute (VRI), Ipoh, Perak, Malaysia
OBJECTIVE
 Define the major characteristics of
each stage in
Biosafety Levels.
 Relate the Risk Groups with the Biosafety Levels.
Prepared by :
Prepared by:
Debbra Marcel, Monoclonal Antibody Section,
MONOCLONAL ANTIBODY SECTION
Veterinary Research Institute (VRI)
WHY DO WE NEED TO HAVE KNOWLEDGE
ABOUT THE BIOSAFETY LEVELS?
 Most of us are always dealing with a potentially harmful
microbe.
 Precautions must be taken in the laboratory to make sure
ourselves and others are not infected.
Ask questions for better understanding…
 Where in the lab would you complete your work?
 What protective equipment and practices would you use?
 How would you contain the microbe to limit contamination or
accidental infection?
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DEFINITION
 What is Biosafety?
- the application of safety precautions that reduce a
laboratorian’s risk of exposure to a potentially
infectious microbe and limit contamination of the work
environment and, ultimately, the community.
 What are Biosafety Levels (BSLs)?
- the level of safety from exposure to infectious agents;
depends on work practices and safety equipment and
facilities
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BIOSAFETY LEVEL
SPECIFIC CONTAINMENT CONTROLS
 Each level are required for the following:
1. Laboratory practices (esp. GLP)
2. Safety equipment (e.g PPE)
3. Facility construction (BSC)
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4 LEVELS OF BIOSAFETY
High risk microbes
EVERY LEVEL MUST
FOLLOW THE STANDARD
MICROBIOLOGICAL
PRACTICES.
Low risk
microbes
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STANDARD MICROBIOLOGICAL
PRACTICES
1.
2.
3.
Not eating, drinking, or applying
cosmetics in the lab
Washing hands after working
with infectious materials and
before leaving the lab
Routinely decontaminating work
surface
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LEVELS OF CONTAINMENT ?
The primary risks: 1. Infectivity
2. Severity of disease
3. Transmissibility
4. The nature of the work conducted
5. Origin of the microbe/agent in question
6. The root of exposure
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BSL-1 (BIOSAFETY LEVEL 1)
• Well-characterized agents with
minimal potential hazard
• Laboratories not necessarily
separated from the general building
• Work typically conducted on open
bench tops
• Specific training in the procedures
conducted in the laboratory and must
be supervised by a scientist with
training in microbiology or a
related science.
• e.g non-pathogenic E. Coli
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BSL-1 (BIOSAFETY LEVEL 1)
1. Laboratory practices


Standard microbiological practices
Work can be performed on an open lab bench
or table. A
2. Safety equipment

Personnel protection equipment (e.g lab
coats, gloves, eye protection) are worn as
needed. B
3. Facility construction



Not required special design on containment
facility
Laboratory doors to separate the working
space with the rest of the facility.
A sink for hand washing
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BSL-2 (BIOSAFETY LEVEL 2)
 builds upon bsl-1.
 the microbes pose moderate hazards
to laboratorians and the environment
 laboratory personnel have specific
training in handling pathogenic
agents
 access to the laboratory is restricted
when work is being conducted
 personnel are supervised by
scientists competent in handling
infectious agents and associated
procedures
 microbes are typically indigeneous and
associated with diseases of varying
severity (e.g Staphylococcus aureus )
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BSL-2 (BIOSAFETY LEVEL 2)
BSL-2 laboratories containment requirements:
1. Laboratory practices

Access to the laboratory is restricted when work is
being conducted.
2. Safety equipment



Appropriate PPE is worn ( lab coats, gloves, eye
protection, face shields, etc.) A
All procedures that can cause infection from
aerosols performed in a biosafety cabinet (BSC)
B
Autoclave/method of decontamination is available
for proper disposals.
3. Facility construction

The laboratory has self-closing doors.

A sink and eyewash are readily available.
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BIOHAZARD !!!
• Mandatory Warning Sign
• Designate Biosafety Level
• Special Entry Procedures
– Immunizations
– PPE
• Contact Information
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BSL-3 (BIOSAFETY LEVEL 3)








builds upon BSL-2.
clinical, diagnostic, teaching, research, or production
facilities.
Work indigenous or exotic agents (serious or
potentially lethal disease) ~ inhalation route
exposure.
Laboratory personnel MUST receive specific
training in handling pathogenic & potentially lethal
agents.
Must be supervised by competent scientists.
BSL-2 + procedures involving the manipulation of
infectious materials must be conducted within
BSCs.
The microbes indigenous/exotic and can cause serious
or potentially lethal disease through respiratory
transmission.
e.g Mycobacterium tuberculosis, the bacteria that
causes tuberculosis.
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BSL-3 (BIOSAFETY LEVEL 3)
1. Laboratory practices
1.

Under medical surveillance and might receive
immunizations for microbes they work with.
Access to the laboratory is restricted and controlled at
all times.
2. Safety equipment


Appropriate PPE must be worn, and respirators
might be required. A
All work with microbes must be within an
appropriate BSC. B
3. Facility construction



A hands-free sink and eyewash near the exit.
The laboratory must have special engineering and
design features directional airflow C (drawing air
from clean areas towards potentially contaminated
areas).
Entrance = two sets of self-closing and locking doors.
C
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BSL-4 (BIOSAFETY LEVEL 4)




builds upon BSL-3 (the highest level of biological safety) = very
few.
The microbes are dangerous and exotic, high risk of aerosoltransmitted infections (frequently fatal and without treatment
or vaccines).
Identical agents must be handled at BSL-4 until sufficient data
obtained.
All laboratory staff and supervisor must:
1. Have specific and thorough training (handling extremely
hazardous infectious agents).
2. Understand the primary and secondary containment
functions of standard and special practices, containment
equipment, and laboratory design characteristics.
3. Competent in handling agents and procedures requiring
BSL-4 containment.

e.g Marburg and Ebola viruses (high individual risk of lifethreatening disease, aerosol transmission, or related agent with
unknown risk of transmission).

c
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BSL-4 (BIOSAFETY LEVEL 4)
1. Laboratory practices

Change clothing before entering, shower upon exiting,
decontaminate all materials before exiting.
2. Safety equipment
•
Must be performed within an appropriate Class III BSC , or
by wearing a full body, air-supplied, positive pressure suit. A
3. Facility construction




The laboratory is isolated and restricted zone.
The laboratory has dedicated supply and exhaust air,
vacuum lines and decontamination systems.
Access to the laboratory is controlled by the laboratory
supervisor (Institutional Policies).
Two types of laboratory providing absolute separation of the
worker from the infectious agents = Suit Laboratory &
Cabinet Laboratory
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SUMMARY
Relation of biosafety levels, practices and equipments to the risk group: BIOSAFETY
LEVEL
LABORATORY
TYPE
Basic: BSL-1
Basic teaching,
research
GMT
Primary health
services;
diagnostic
services,
research
GMT plus
protective
clothing,
biohazards sign
Open bench plus
BSC for aerosols
Containment:
BSL-3
Special
diagnostic
services,
research
Level 2 + special
clothing, access
control, directed
airflow
BSC and/or other
primary devices for
all activities
Max.
containment:
BSL-4
Dangerous
pathogen units
Level 3 + airlock
entry, shower exit,
special waste
disposal
Class III BSC, or
positive pressure
suites with class II
BSCs, double ended
autoclave
Basic: BSL-2
LABORATORY
PRACTICES
SAFETY
EQUIPMENT
RISK
GROUP
None; open bench
work
EXAMPLE
1
canine hepatitis, nonpathogenic E. coli, some
cell cultures & noninfectious bacteria
2
Staphylococcus aureus,
Lyme disease,
Salmonella, mumps,
measles
3
Mycobacterium
tuberculosis, Brucella
sp., rabies virus,
Chikungunya
4
Marburg virus, Ebola
virus, Nipah virus
GMT = Good microbiological techniques ; BSC = Biosafety cabinet
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REFERENCES:
 CDC website: http://www.cdc.gov/
(e-learning, quick lessons, many interactive resources)
 Wikipedia: http://en.wikipedia.org/wiki/Biosafety_level

Biosafety in Microbiological and Biomedical Laboratories (BMBL)
5th Edition: e-book
http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
Prepared by :
Debbra Marcel, Monoclonal Antibody Section,
Veterinary Research Institute (VRI)