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Laboratory Hazards
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Biosafety Level -1 (BSL-1)

● Biosafety level 1 is the lowest

safety level.
● These laboratories provide
general space in which work is
done with viable agents that are
not associated with disease in
healthy adults.
Uses
● Biosafety Level-1 is commonly used
while performing tests on microbial
agents that are not known to cause
diseases in individuals.
● These laboratories include the
laboratories used for teaching
purposes in colleges and training
centers.
Safety practices
● There are NO specific safety practices
required for BSL-1.

Safety equipment
● Special containment devices like the
Biosafety Cabinets are NOT required for
BSL-1.
● In order to prevent the contamination of
personal clothing, protective laboratory
coats, gowns, or uniforms are
recommended.
● While conducting tests with a high possibility
of aerosol formation, protective eyewear can
be used.
Organisms
● Agrobacterium radiobacter,
Aspergillus niger, Bacillus
thuringiensis, Escherichia coli strain
K12, Lactobacillus acidophilus,
Micrococcus leuteus, Neurospora
crassa, Pseudomonas fluorescens,
Serratia marcescens.
● However, the requirement of the
biosafety level might differ depending
on the risk assessment of the
pathogen.
Biosafety Level-2 (BSL-2)

• Biosafety level-2 laboratories are the

laboratories that are used for the tasks
involving microbial agents of moderate
potential hazards to the laboratory
personnel, the environment, and the
agent.
• The infectious agents or the toxins might
pose a moderate danger if accidentally
inhaled, swallowed, or exposed to the
skin.
Requirements

• Access into the laboratory is limited

while BSL-2 experiments are in progress.
• The annual inspection of the laboratories
is also an important part of the BSL-2
requirements. These might include
changing the filters or replacement of
some devices.
• The work is mostly conducted on
sterilized bench tops except for some
processes that might form aerosols. The
latter is conducted in safety cabinets.
Safety Practices

• People with increased risk of acquiring

infections like the immune-compromised and
pregnant individuals should not be allowed
to enter the BSL-2 laboratories while the
laboratories are at work.
• An annual review of the BSL-2 manual should
be done to update the guidelines.
• Documented policies and procedures should be
established that limit the entrance to individuals
who know of the potential hazards and are
appropriately trained.
• A biohazard symbol is placed on pieces of
equipment where biohazardous materials are
used or stored.
Safety Equipment
• Protective coats
• The laboratory design should be made such that it
can be easily cleaned and decontaminated with
minimum nooks and corners.
• The laboratory doors should be closed whenever
work with hazardous biomaterials is conducted.
• An autoclave must be available.

Uses
● Biosafety level-2 laboratories are mostly used for
routine analysis and culture of moderately
hazardous agents.
● Besides, some of the laboratories used for teaching
and training purposes are also BSL-2 laboratories.
Organisms

• The organisms that require BSL-2

laboratories include the pathogenic
strains of E. coli, Staphylococcus,
Salmonella, Plasmodium falciparum,
Toxoplasma, and Herpes Simples
Viruses.
• The allocation of organisms to the
laboratories, however, might differ
depending on the risk assessment.
Biosafety Level-3 (BSL-3)
● Work is performed with agents that may cause
severe or potentially lethal disease through
inhalation or aerosol formation, to the personnel,
and may even contaminate the environment.
● The tasks potential for infection by aerosols is
high, and the disease may have lethal
consequences.
● Working in such laboratories require laboratory
personnel with specific training in handling
pathogenic and potentially lethal agents, along
with supervisors scientists competent in handling
infectious agents and associated procedures.
Requirements

● These laboratories should be certified for use before

initial operation and on an annual schedule or after a
program change, renovation, or replacement of system
components that may affect the operating environment of
the laboratory.
● BSL-3 laboratories are also called the containment
laboratory as they require containment equipment to
protect the personnel, the microbial agent, and the
environment.
● The requirements for BSL-3 include all the requirements
of the BSL-1 and BSL-2 laboratories, along with some
additional design features and special equipment.
Standard Microbiological Practices

● The entry to the BSL-3 laboratories is limited to

individuals with appropriate training in handling BSL-
3 organisms.
● All the procedures to be conducted in the BSL-3 must be
conducted within a biosafety cabinet to prevent the
exposure of the aerosols to the laboratory personnel.
● Personnel working in the laboratory must wear personal
protective equipment before entering the laboratory and
then remove them before leaving.
● The work surfaces and sinks should be decontaminated
once every work shift or after any spills or splashes.
● The BSL-3 laboratories should be separated from the
general traffic in a building to limit entry into the
laboratories at all times.
Safety Practices
● The doors of the BSL-3 laboratories are closed at all
times with appropriate BSL-3 signs outside the suite, along
with a universal biohazard sign and emergency contact
information.
● Laboratory personnel must have medical surveillance
and offered appropriate immunizations for agents
handled or potentially present in the laboratory.
● A laboratory-specific biosafety manual, which is available
and accessible to all, must be prepared and adopted as a
policy.
● No work in the BSL-3 laboratories should be
conducted on an open bench or an open vessel.
Uses
● BSL-3 laboratories are used for clinical, diagnostic, teaching,
research, or production facilities.
● These laboratories are used for the handling and
manipulation of highly infectious agents that prose direct
severe effects on the health of the personnel.
● These are used for the studies regarding the effects of
infectious agents and various toxins and their effects.

Organisms
● The pathogens that require BSL-3 laboratories include HIV,
Mycobacterium tuberculosis, SARS, Rabies Virus, etc.
● The placement of the organisms in different Biosafety levels,
however, might defer and should also be determined after risk
assessment.
Biosafety Level 4 (BSL-4)

• The BSL-4 laboratories are often used while

handling and manipulating Risk Group 4
pathogens that are extremely dangerous,
with no known vaccines or therapies, and
require extreme precautions during work.
Requirements

• The requirements of BSL-4 laboratories are extensive with

specific laboratory design, training procedures, and
highly protective equipment and personal gears.
• These laboratories should be certified for use before initial
operation and on an annual schedule or after a program
change, renovation, or replacement of system components
that may affect the operating environment of the laboratory.
• BSL-4 laboratories are also termed the maximum
containment laboratories as they have secondary barriers
to prevent hazardous materials from escaping into the
environment.
• The BSL-4 laboratories should follow the requirements of all
BSL-1, BSL-2, and BSL-3, along with additional specific
precautions.
Standard microbiological practices

• No work conducted within the BSL-4 should

be done on an open bench or an open
vessel.
• Only people that are trained in handling the
BSL-4 organisms and the equipment in the
laboratory should be allowed into the
laboratory.
Safety equipment

• Class III biological safety cabinet.

• Walls, floors, and ceilings of the
laboratories must form a sealed internal
shell which facilitates fumigation and is
animal and insect-proof.
• A double-doored autoclave is placed for
decontaminating materials passing out of
the facility.
• The exhaust air from the facility is filtered
through HEPA filters before being
discharged to the outside so as to prevent
its entry into occupied buildings and air
intakes.
Uses
● BSL-4 laboratories are used for diagnostic and research
work on easily transmitted pathogens, causing fatal
diseases.
● These laboratories are used for new and unknown
pathogenic microbes, for which no vaccines or
therapies are available.
● They are also used for clinical and production facilities
that require highly sophisticated techniques and
advanced processes.

Organisms
● The BSL-4 level pathogens include the risk group IV
organisms like Ebola virus, SARS-CoV-2, Hemorrhagic
viruses, etc.