BIOSAFETY LEVELS
ALBIN BENNY
Biosafety
• Containment principles, technologies and
practices that are implemented to prevent the
unintentional exposure to biological material
or their accidental release (adapted from
WHO, 2006).
• Biosafety is the prevention of large-scale loss
of biological integrity, focusing both
on ecology and human health.
• Biosafety is used to protect from harmful
incidents.
• Many laboratories handling biohazards employ
an ongoing risk management assessment and
enforcement process for biosafety.
• Failures to follow such protocols can lead to
increased risk of exposure to biohazards or
pathogens.
• When biological warfare or new, currently
hypothetical, threats (i.e., robots, new
artificial bacteria) are considered, biosafety
precautions are generally not sufficient. The
new field of biosecurity addresses these
complex threats.
Containment
• The term containment is used to describe the
safe work practices in handling infectious agents
to reduce exposure to laboratory personnel and
others.
• Types of containment
• • Biological containment
• • Physical containment
Biological containment (BC)
• Biocontainment is a component of biorisk
management. The overall objective of
biocontainment is to confine an infectious
organism or toxin, thereby reducing the
potential for exposure to laboratory workers
or persons outside the laboratory, and the
likelihood of accidental release to the
environment.
Physical Containment (PC)
• Physical containment helps to
confine the pathogenic organisms
being handled and prevent exposure
to personnel.
• Physical containment is achieved through the
use of laboratory practices, containment
equipment, personal protective equipment,
and laboratory and facility design.
BIOSAFETY LEVELS
• A biosafety level is the level of the
biocontainment precautions, required
to undertake while handling
dangerous biological agents in an
enclosed facility.
Biosafety level 1 (BSL-1)
• Biosafety level 1 (BSL-1) is suitable for work with
well-characterized agents which do not cause
disease in healthy humans.
• In general, these agents should pose minimal
potential hazard to laboratory personnel and the
environment.
• At this level, precautions are limited relative to
other levels.
• Laboratory personnel must wash their hands upon
entering and exiting the lab.
• Research with these agents may be
performed on standard open laboratory
benches without the use of special
containment equipment.
• Eating and drinking are generally prohibited
in laboratory areas.
• Potentially infectious material must be
decontaminated before disposal, either by
adding a chemical such
as bleach or isopropanol or by packaging for
decontamination elsewhere.
• Personal protective equipment is only required for
circumstances where personnel might be exposed to
hazardous material.
• BSL-1 laboratories must have a door which can be locked
to limit access to the lab.
• It is not necessary for BSL-1 labs to be isolated from the
general building.
• This level of biosafety is appropriate for work with several
kinds of microorganisms including non-pathogenic strains
of Escherichia coli and Staphylococcus, Bacillus
subtilis, Saccharomyces cerevisiae and other organisms not
suspected to contribute to human disease.
• Due to the relative ease and safety of maintaining a BSL-1
laboratory, these are the types of laboratories generally
used as teaching spaces for high schools and colleges.
Biosafety level 2
• Biosafety level 2 is suitable for work involving
agents of moderate potential hazard to
personnel and the environment. This includes
various microbes that cause mild disease to
humans, or are difficult to contract via aerosol
in a lab setting.
• Laboratory personnel have specific training in
handling pathogenic agents and are directed
by scientists with advanced training.
• Access to the laboratory is limited when work
is being conducted.
• Extreme precautions are taken with
contaminated sharp items.
• Certain procedures in which
infectious aerosols or splashes may be created
are conducted in biological safety cabinets or
other physical containment equipment.
Biosafety level 3
• Biosafety level 3 is appropriate for work
involving microbes which can cause serious
and potentially lethal disease via the
inhalation route. This type of work can be
done in clinical, diagnostic, teaching, research,
or production facilities.
• All laboratory personnel are provided medical
surveillance and offered relevant immunizations
(where available) to reduce the risk of an accidental
or unnoticed infection.
• All procedures involving infectious material must be
done within a biological safety cabinet.
• Laboratory personnel must wear solid-front
protective clothing (i.e. gowns that tie in the back).
This cannot be worn outside of the laboratory and
must be discarded or decontaminated after each use.
• A laboratory-specific biosafety manual must be
drafted which details how the laboratory will operate
in compliance with all safety requirements.
Biosafety cabinet
• A biosafety cabinet (BSC)—also called
a biological safety cabinet or microbiological
safety cabinet—is an enclosed,
ventilated laboratory workspace for safely
working with materials contaminated with (or
potentially contaminated with) pathogens
requiring a defined biosafety level. Several
different types of BSC exist, differentiated by the
degree of biocontainment they provide. BSCs first
became commercially available in 1950.
• In addition, the facility which houses the BSL-3 laboratory
must have certain features to ensure appropriate
containment.
• The entrance to the laboratory must be separated from areas
of the building with unrestricted traffic flow.
• Additionally, the laboratory must be behind two sets of self-
closing doors (to reduce the risk of aerosols escaping).
• The construction of the laboratory is such that it can be
easily cleaned.
• Carpets are not permitted, and any seams in the floors,
walls, and ceilings are sealed to allow for easy cleaning and
decontamination.
• Additionally, windows must be sealed, and a ventilation
system installed which forces air to flow from the "clean"
areas of the lab to the areas where infectious agents are
handled. Air from the laboratory must be filtered before it
Biosafety level 4
• Biosafety level 4 (BSL-4) is the highest level of
biosafety precautions, and is appropriate for
work with agents that could easily be aerosol-
transmitted within the laboratory and cause
severe to fatal disease in humans for which
there are no available vaccines or treatments.
• BSL-4 laboratories are generally set up to be
either cabinet laboratories or protective-suit
laboratories.
• The cabinets themselves are required to have
seamless edges to allow for easy cleaning.
• Additionally, the cabinet and all materials within must
be free of sharp edges in order to reduce the risk of
damage to the gloves.
• In a protective-suit laboratory, all work must be done
in a class II biosafety cabinet by personnel wearing
a positive pressure suit.
• In order to exit the BSL-4 laboratory, personnel must
pass through a chemical shower for decontamination,
then a room for removing the positive-pressure suit,
followed by a personal shower.
• Entry into the BSL-4 laboratory is restricted to trained
and authorized individuals, and all persons entering
and exiting the laboratory must be recorded.
• As with BSL-3 laboratories, BSL-4 laboratories must be
separated from areas that receive unrestricted traffic.
• Additionally airflow is tightly controlled to ensure that
air always flows from "clean" areas of the lab to areas
where work with infectious agents is being performed.
• The entrance to the BSL-4 lab must also
employ airlocks to minimize the possibility that
aerosols from the lab could be removed from the lab.
• All laboratory waste, including filtered air, water, and
trash must also be decontaminated before it can leave
the facility.
• Biosafety level 4 laboratories are used for diagnostic
work and research on easily transmitted pathogens
which can cause fatal disease. These include a number
of viruses known to cause viral hemorrhagic
fever such as Marburg virus, Ebola virus, Lassa virus,
and Crimean-Congo hemorrhagic fever. Other
pathogens handled at BSL-4 include Hendra virus,
Nipah virus, and some flaviviruses.
• Additionally, poorly characterized pathogens which
appear closely related to dangerous pathogens are
often handled at this level until sufficient data are
obtained either to confirm continued work at this
level, or to permit working with them at a lower level.
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