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Containment

• The term containment describes safe methods for managing infectious materials in
the laboratory environment where they are being handled or maintained.
• The purpose of containment is to reduce or eliminate exposure to potentially
hazardous agents.
• Exposure could involve not only laboratory workers but other individuals working
close by and the outside environment.
Biosafety Levels • The three elements of containment include:

1. laboratory practice and technique

2. safety equipment
3. facility design

The risk assessment of the work to be done with a specific agent will determine the
appropriate combination of these elements.

Primary Containment Secondary Containment

• Secondary containment is the
protection of the environment external
• Primary containment is the protection of personnel and the immediate to the laboratory from exposure to
laboratory environment from exposure to infectious agents. infectious materials.

• Primary containment is provided by both good microbiological technique and the • Secondary containment is provided by a
use of appropriate safety equipment. combination of facility design and
operational practices.

For example, the use of vaccines may provide an increased level of personal
protection. Personal protective equipment such as gowns, masks, and gloves and • Ventilation systems, controlled access,
biological safety cabinets offer protection when used properly in conjunction with airlocks, and other facility design
good laboratory techniques. features must be part of any biosafety
program.
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Biological Safety Cabinets

Class I BSC
• Safety equipment includes biological safety
cabinets (BSCs), enclosed containers, and other
engineering controls designed to remove or • Class I BSC is the first designed and
minimize exposures to hazardous biological simple Biological Safety Cabinet
materials. (BSC) which provides personnel
and environmental protection but
not product production (as
• The biological safety cabinet (BSC) is the principal unsterilized room air is drawn over
device used to provide containment of infectious
splashes or aerosols generated by many the work surface).
procedures.
• Class I BSCs are suitable for work
• There are three types of biological safety cabinets with Risk Group 1 (RG1), Risk
used in microbiological and biomedical Group 2 (RG2), and Risk Group 3
laboratories - Class I, Class II, and Class III. (RG3) biological material.

Class III BSC

Class II BSC • Class III BSC provides the highest level of personnel
protection and is used for Risk Group 4 agents. It is
suitable for work in Biosafety Level 3 and 4
• A Class II BSC is a ventilated cabinet, laboratories.
which provides personnel, product and
environmental protection. • This type of cabinet is totally enclosed and is tested
under pressure to ensure that no particles can leak
from it into the room.
• It is commonly found in clinical and • Operator access the work surface by means of heavy-
research laboratories working with duty rubber gloves which form part of the cabinet.
infectious agents in Risk Groups 2, 3
and 4 or with tissue culture. • Airflow is maintained by a dedicated exhaust system
exterior to the cabinet, which keeps the cabinet
interior under negative pressure.
• About 90% of all biosafety cabinets
installed are Type A2 cabinets • The Class III cabinet may be connected to a double-
door autoclave used to decontaminate all materials
entering or exiting the cabinet.
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• Facility design and construction contribute to the

laboratory workers' protection, provide a barrier Secondary Barriers
The Facility as a Barrier to protect persons outside the laboratory, and • The recommended secondary barrier(s) will depend on the risk of
protect people and animals in the community transmission of specific agents.
from infectious agents which may be accidentally
released from the laboratory.
• When the risk of infection by exposure to an infectious aerosol is present,
• Laboratory management is responsible for higher levels of primary containment and multiple secondary barriers may
providing facilities that are commensurate become necessary to prevent infectious agents from escaping into the
with the laboratory's function and with the environment .
recommended biosafety level for the agents
being manipulated. Such design features include:
• Specialized ventilation systems to ensure directional air flow
• A variety of experts should be part of the • Air treatment systems to decontaminate or remove agents from exhaust air
design team for any new facility. These include
biosafety professionals, HVAC engineers and • Controlled access zones
animal care professionals. • Airlocks as laboratory entrances (as shown in this image)
• Separate buildings or modules to isolate the laboratory

Biosafety Levels Biosafety Level 1

As the lowest of the four, biosafety level 1 applies to laboratory settings in which
• A biosafety level (BSL), or pathogen/protection level, is a set of personnel work with low-risk microbes that pose little to no threat of infection in
biocontainment precautions required to isolate dangerous biological healthy adults. An example of a microbe that is typically worked with at a BSL-1 is a
agents in an enclosed laboratory facility. nonpathogenic strain of E. coli.
• This laboratory setting typically consists of research taking place on benches without
the use of special contaminant equipment. A BSL-1 lab, which is not required to be
• The levels of containment range from the lowest biosafety level 1 isolated from surrounding facilities, houses activities that require only standard
(BSL-1) to the highest at level 4 (BSL-4). microbial practices, such as:

• Mechanical pipetting only (no mouth pipetting allowed)

• Biological Safety Levels (BSL) are a series of protections relegated to
autoclave-related activities that take place in particular biological • Safe sharps handling
labs. They are individual safeguards designed to protect laboratory
personnel, as well as the surrounding environment and community. • Avoidance of splashes or aerosols
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Biosafety Level 2
• Daily decontamination of all work surfaces when work is complete • This biosafety level covers laboratories that work with agents associated with
human diseases (i.e. pathogenic or infections organisms) that pose a
• Hand washing moderate health hazard. Examples of agents typically worked with in a BSL-2
• Prohibition of food, drink and smoking materials in lab setting include equine encephalitis viruses and HIV, as well as Staphylococcus aureus
• Personal protective equipment, such as; eye protection, gloves and a lab coat or gown (staph infections).
• Biohazard signs
• BSL-1 labs also requires immediate decontamination after spills. Infection materials are • BSL-2 laboratories maintain the same standard microbial practices as BSL-1
also decontaminated prior to disposal, generally through the use of an autoclave. labs, but also includes enhanced measures due to the potential risk of the
aforementioned microbes. Personnel working in BSL-2 labs are expected to
take even greater care to prevent injuries such as cuts and other breaches of
the skin, as well as ingestion and mucous membrane exposures.

• In addition to BSL 1 expectation, the following practices are required in a BSL 2

lab setting:

• Appropriate personal protective equipment (PPE) must be worn, including lab coats
and gloves. Eye protection and face shields can also be worn, as needed. Biosafety Level 3
• All procedures that can cause infection from aerosols or splashes are performed • Again building upon the two prior biosafety levels, a BSL-3 laboratory typically
within a biological safety cabinet (BSC). includes work on microbes that are either indigenous or exotic, and can cause
serious or potentially lethal disease through inhalation. Examples of microbes
worked with in a BSL-3 includes; yellow fever, West Nile virus, and the bacteria
• An autoclave or an alternative method of decontamination is available for proper that causes tuberculosis.
disposals.

• The laboratory has self-closing, lockable doors. • The microbes are so serious that the work is often strictly controlled and
registered with the appropriate government agencies. Laboratory personnel
are also under medical surveillance and could receive immunizations for
• A sink and eyewash station should be readily available. microbes they work with.

• Biohazard warning signs

• Common requirements in a BSL-3 laboratory include:
• Access to a BSL-2 lab is far more restrictive than a BSL-1 lab. Outside personnel, or
those with an increased risk of contamination, are often restricted from entering • Standard personal protective equipment must be worn, and respirators might
when work is being conducted. be required
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Biosafety Level 4
• Solid-front wraparound gowns, scrub suits or coveralls are often required
• BSL-4 labs are rare. However some do exist in a small number of places
around the world. As the highest level of biological safety, a BSL-4 lab
• All work with microbes must be performed within an appropriate BSC
consists of work with highly dangerous and exotic microbes. Infections
caused by these types of microbes are frequently fatal, and come without
• Access hands-free sink and eyewash are available near the exit treatment or vaccines. Two examples of such microbes include Ebola and
Marburg viruses.

• Sustained directional airflow to draw air into the laboratory from clean areas
towards potentially contaminated areas (Exhaust air cannot be re-circulated) • In addition to BSL-3 considerations, BSL-4 laboratories have the following
containment requirements:

• A self closing set of locking doors with access away from general building
corridors • Personnel are required to change clothing before entering, shower upon
exiting

• Access to a BSL-3 laboratory is restricted and controlled at all times.

• Decontamination of all materials before exiting

Risk Assessment
• Personnel must wear appropriate personal protective equipment from prior
BSL levels, as well as a full body, air-supplied, positive pressure suit Assessing the risk for acquiring a laboratory associated infection is affected by the
following factors:
• A Class III biological safety cabinet • Pathogenicity
• Route of Transmission
• A BSL-4 laboratory is extremely isolated—often located in a separate building • Agent Stability
or in an isolated and restricted zone of the building. The laboratory also • Infectious Dose
features a dedicated supply and exhaust air, as well as vacuum lines and
decontamination systems. • Susceptibilty
• Concentration and Volume
• Knowing the difference in biosafety lab levels and their corresponding safety • Origin
requirements is imperative for anyone working with microbes in a lab setting.
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Risk Assessment
• Pathogenicity Risk Assessment
• The greater the pathogenicity of the infectious or suspected infectious agent, the
more severe is the potentially acquired disease, and so the higher is the risk.
• For example: • Route of Transmission
• Agents transmitted by the aerosol route have caused the most laboratory
infections, versus agents transmitted parenterally or by ingestion. When
• Since Staphylococcus aureus rarely causes a severe or life-threatening disease in a planning work with an unknown agent with an uncertain mode of
laboratory situation, it is assigned to BSL-2.
transmission, the potential for aerosol transmission must be considered due
to the higher risk.
• Ebola, Marburg, and Lassa fever viruses cause diseases with high mortality rates and
have no vaccines or treatment, so BSL-4 is the appropriate level to work with those • Agent Stability
viruse.
• Desiccation, exposure to sunlight or ultraviolet light, exposure to chemical
disinfectants and other factors can affect the agent's stability in the
• Work with human HIV and hepatitis B virus is done at BSL-2 because they are not environment.
transmitted by the aerosol route, even though potentially lethal disease can result from
exposure. For hepatitis B, there is also an effective vaccine available.

Risk Assessment Risk Assessment

• Infectious Dose • Concentration and Volume
• Infectious dose can vary from one to hundreds of thousands of units. • The concentration is the number of infectious organisms per unit volume.
Higher concentrations increase the risks of working with that agent. Working
• Susceptibility with large volumes of concentrated infectious material also increases the
• The infectious dose is affected by the individual's resistance, so a laboratory worker's risks, since additional handling of the materials is often required.
immune status is directly related to his/her susceptibility to disease when working
with an infectious agent. Thus, susceptibility may be greater than in a healthy person
for persons who are pregnant, have undergone surgery, are receiving immune- • Origin
suppressent medications (including steroids), or who have systemic infectious
diseases. • Origin may refer to geographic location (e.g., domestic or foreign); host (e.g.,
infected or uninfected human or animal); or nature of source (potential
zoonotic or associated with a disease outbreak).
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Personal Protection Equipment

• Personal protective equipment (PPE)
is often used in combination with
biological safety cabinets and other
devices that contain the agents,
animals, or materials being handled.
Personal Protective Equipment
• It may be difficult or impractical to
work in biological safety cabinets in
some situations; in this instance,
personal protective equipment may
form the primary barrier between
personnel and the infectious
materials.

General types of personal protective Gloves

equipment
• Hand protection, e.g., gloves Check throughout task for tears, punctures,
• Body protection discolouration, stiffening, softening
• Eye protection Change after 1 use, if feasible
• Face protection
Protect the skin under the gloves
• Hearing protection
• Respiratory protection Use double gloves whenever you may need to
• not addressed in this presentation change gloves in contaminated area
• Fall protection
• not addressed in this presentation Avoid touching clean areas with contaminated
• Combinations, ensembles gloves

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Body protection, various types Eye protection

• Coveralls • Safety glasses should be worn throughout process.
• Protects most of body
• Legs, arms, torso • Use goggles for greater particle protection and chemical splashes
• May also include head and feet • Ensure good fit – contact at nose and behind ears
• Different styles to protect against: • Application against:
• Particulate, e.g., dust • Chemical spray or splash
• Chemical, e.g., spray or splash • Dust
• Thermal, e.g., flash fire • Impact
• Aprons • Gas-tight models (e.g., formaldehyde vapor)
• Overhead work (e.g., falling debris that could get past standard safety
• Sleeves glasses).

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Face protection, the face shield Mind the gap

• A face shield is considered “secondary” protection; a face shield should not • Gap between protective garments
be used alone, e.g., without safety glasses or goggles, depending on the
hazard. • Gloves and coveralls
• Boots and coveralls
• Application
• Impact • Respirator and coveralls
• Chemical spray or splash • Addressing the gap
• Radiant energy • Elastic cuffs (e.g., ankle, wrist, and hood)
• Other • ChemTape
• Head band or hardhat mounting options • Over-sleeves
• A full face respirator is typically considered equivalent protection to a goggles • Integral equipment (e.g., gloves part of suit)
and face shield combination.

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