Lab. Biosafety Laws!

Biosafety
levels & Biosafety Cabinets
 What is Biosafety!
 Biosafety is the prevention of large-scale loss of
biological integrity,
 It focusing both on ecology and human health.
 These prevention mechanisms include conduction of
regular reviews of the biosafety in laboratory settings, as
well strict guidelines to follow..
 Biosafety is used to protect from harmful incidents.
 Failures to follow such protocols can lead to increased risk
of exposure to biohazards or pathogens.
 Biosafety in agriculture, chemistry, medicine etc. require the
application of biosafety principles.
 Biosafety levels refers to the stringency of bio-
containment precautions deemed necessary for the
laboratory work with infectious materials.
 BIOSAFETY LEVELS
 A biosafety level is a set of bio-containment
precautions required to isolate dangerous biological
agents in an enclosed laboratory facility.
 The levels of containment range from the
lowest biosafety level-I to the highest biosafety level-
IV.
 There are total IV biosafety levels i-e. BSL-I, BSL-II,
BSL-III & BSL-IV, respectively.
 WHY WE NEED BIOSAFETY?
 Lab. has hazards of processing
infectious agents (Mirobs, i-e. Bacteria,
Viruses etc.).
 Accidental threats to workers and the
controlled environment.
 To have adherence with safety
regulations while dealing with the
highly infectious agents (Corona Virus,
Ebula Virus, Tuberculosis Bacteria
etc.).
 BIOSAFETY LEVEL # I (BSL-I)
 BSL-I labs microbes not known consistently to cause
disease in healthy adults,
 and present minimal potential hazard to lab and the
controlled environment.
 Example: Non-pathogenic strain of Escherichia coli
bacteria etc.
BIOSAFETY LEVEL # II (BSL-II) & PRACTICES
 Standard microbiological practices are followed.
 Work can be performed on an open table or bench in a
laboratory.
 PPE (Personal Protective Equipment) needed.
 Personal protective equipment is protective clothing,
helmets, goggles, garments or other equipments
designed to protect the body from injury/ infection etc.
 Sink – hand washing.
 Lab./ laboratory doors separate.
BIOSAFETY LEVEL # II (BSL-II) &
PRACTICES
 BIOSAFETY LEVEL # II PRACTICES
 Biosafety Level- II (BSL-II) laboratories are used to study
moderate-risk infectious agents,
 or toxins that pose a moderate danger, if accidentally
inhaled, swallowed, or exposed to the skin.
 At this level, all precautions used at Biosafety Level-I are
followed, and some additional precautions are taken.
 BSL-II differs from BSL-I in that:
 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.
 Biosafety level-II is suitable for work involving
hepatitis A, B, C viruses, human immunodeficiency
virus (HIV), pathogenic strains of Escherichia coli,
Staphylococcus, Salmonella, Plasmodium, Toxoplasma
etc.
 Prions, the infectious agents that transmit prion
diseases such as vCJD (Variant Creutzfeldt-Jakob
disease) may be handled under Biosafety Level-II or
higher.
 BIOSAFETY LEVEL # III (BSL-III) &
PRACTICES
 Biosafety level-III 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.
 The precautions undertaken in BSL-I & BSL-II labs are
followed, as well as additional measures including:
 All laboratory personnel are provided medical surveillance
and offered relevant immunizations 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.
 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 LEVEL # IV (BSL - IV) &
PRACTICES
 Biosafety level # IV (BSL-IV) 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-IV laboratories are generally set up to be either
cabinet laboratories or protective-suit laboratories.
 In cabinet laboratories, all work must be done within a
class III biosafety cabinet.
 Materials leaving the cabinet must be decontaminated by
passing through an autoclave or a tank of disinfectant.
 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 order to exit the BSL-IV laboratory, personnel
must pass through a chemical shower for
decontamination followed by a personal shower.
 Entry into the BSL-IV laboratory is restricted to trained
and authorized individuals,
 and all persons entering and exiting the laboratory
must be recorded to take a proper care.
 BSL IV 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-IV lab must also employ airlocks
to minimize the possibility that aerosols from the lab
could be removed.
 All laboratory waste, including filtered air, water, and
trash must also be decontaminated before it can leave
the facility.
 Biosafety level-IV laboratories are used for diagnostic
work and research on easily transmitted pathogens
which can cause fatal disease.
 These include a number of viruses known such as Ebola
virus, Nipah virus, Corona virus etc.
 BIOSAFETY CABINET (BSC) & PRACTICES
 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 pathogens
requiring a defined biosafety level.
 Several types of BSC exist,
differentiated by the degree of
biocontaminant they provide.
 BSCs first became commercially
available in 1950.
 Safety Cabinet protects both the specimen and the user from
biological contamination.
 It provide personnel, environmental and product protection.
 Particulate-free air is passed down from the top of the hood,
across the work surface, and then captured before entering a
worker's breathing zone.
 The air is then filtered before being exhausted, usually back into
the laboratory.
 Three kinds of biological safety cabinets designated as Class I, II &
III are available to meet varying research and clinical needs.
 All of the types of biological safety cabinets uses high efficiency
particulate air (HEPA) “at least 99.95%” filters or ultra-low
penetration air (ULPA) “at least 99.999%” filters in their exhaust
and/ or supply systems.
 Purposes of Biosafety Cabinets
(BSCs)
 The primary purpose of a BSCs is to serve as a means to
protect the laboratory worker and the surrounding
environment from pathogens.
 All exhaust air is HEPA filtered as it exits the biosafety
cabinet, removing harmful bacteria and viruses.
 Neither are most BSCs safe for use as fume hoods.
 Likewise, a fume hood fails to provide the environmental
protection that HEPA filtration in a BSC would provide.
 However, most classes of BSCs have a secondary
purpose to maintain the sterility of materials inside.
 CLASS # 1 BIOSAFETY CABINET (BSC)
 Class I cabinets provide personnel and environmental
protection but no product protection.
 In fact, the inward flow of air can contribute to
contamination of samples.
 These BSCs are commonly used to enclose specific
equipment like centrifuges or a procedure that potentially
generate aerosols.
 BSCs of this class are either ducted i-e. connected to the
building exhaust system or unducted i-e. recirculating
filtered exhaust back into the laboratory.
 CLASS # I BIOSAFETY
CABINET
A: Front Opening
B: Sash
C: Exhaust HEPA
D: Contaminated Air

[Sash: A movable or fixed

window on the front that
protects workers from the
active work area]
 CLASS # II BIOSAFETY CABINET
 Class II Biological Safety Cabinets provide personnel,
environmental and the product (sample) protection as well.
 The Class II cabinet recirculates approximately 70% of the air
through a supply filter,
 and exhausts approximately 30% of the air through an
exhaust filter to the room.
 The exhaust is directed to the outside through a facility
exhaust system.
 Class II Biological Safety Cabinets are widely used in
toxicology laboratories and similar applications where
chemical effluent is present and clean air is essential.
CLASS # II BIOSAFETY CABINET

A: Font Opening
B: Sash
C: Exhaust HEPA filter
D: Supply HEPA filter
E. Positive pressure plenum
F: Negative pressure plenum
CLASS # III BIOSAFETY CABINET
 The Class III cabinet, generally only installed in maximum
containment laboratories,
 And specifically designed for work with BSL-4 pathogenic
agents, providing maximum protection.
 The enclosure is gas-tight, and all materials enter and
leave through a dunk tank or double-door autoclave.
 Gloves attached to the front prevent direct contact with
hazardous materials.
 Class III cabinets are sometimes called gloves boxes.
 CLASS # III BIOSAFETY
CABINET

A: Glove ports with O-ring for

attaching arm-length gloves to
cabinet
B. Sash
C. Exhaust HEPA filter
D. Supply HEPA filter
E. Double-ended autoclave
or pass-through box
 Positive Pressure
Biosafety Cabinet
Personnel Suits