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Biosafety Training
Module 3: Biocontainment
University Safety & Assurances
Biological Safety Program

Biosafety Levels
BSL- • Biosafety Level defines type
4 of containment, equipment,
PPE, and laboratory
BSL-3 practices implemented in to
work with a biological agent.
BSL-2 • It is not tied directly to the
risk group of an agent.
BSL-1

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Containment Level (Biosafety Level)

• A risk assessment is the most accurate means of

determining the appropriate containment for the work you
plan to do, not what a company says is the appropriate BSL
• Biosafety level is a containment level- lab design, practices,
procedures, equipment
• There are many guidelines regarding biosafety levels of
containment, but the general features for each level of
containment are standard.
• If you are not a PI, the containment level for your lab has
already been designated
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BSL-1 and BSL-2

• When is BSL-1 appropriate?
• Suitable for work involving well-characterized agents not known to
consistently cause disease in immunocompetent adult humans, and
present minimal potential hazard to laboratory personnel and the
environment (BMBL, 5th edition).
• Examples: E.coli K-12, Saccharomyces cerevisiae
• When is BSL-2 appropriate?
• BSL-2 is suitable for work involving agents that pose moderate hazards
to personnel and/ or the environment.
• Differences from BSL-1: training requirements, laboratory access,
engineering controls, PPE requirements
• Examples: Staphylococcus aureus, Pseudomonas aeruginosa

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BSL-2+ Laboratories
• Not an official containment designation.
• BSL-2 labs in which personnel use some BSL-3 practices, such as working
exclusively in the biosafety cabinet, wearing respirators or enhanced PPE.
• Required for work with: Escherichia coli (shiga toxin producing strains), Hepatitis
B Virus (HBV), Hepatitis C Virus (HCV), Human Immunodeficiency Virus (HIV),
Simian Immunodeficiency Virus, Zika virus, Vaccinia virus (or vector), Rabies
virus, Influenza virus (except for pandemic strains- must be worked with in BSL-
3), Infectious Prions, Listeria monocytogenes, Lymphocytic Choriomeningitis
Virus (LCMV), Macaque tissue, Oncogenes used in viral vectors, Salmonella
typhi, Shigella dysenteriae (Type 1), and/ or Toxoplasma gondii
• Additionally, work with any of the above materials requires occupational health
screening and routine medical surveillance at the expense of the laboratory PI.

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BSL-1 and BSL-2 Minimum Criteria

BSL-1 BSL-2 BSL-2+/ enhanced
Primary Biosafety Cabinet that is BSC is currently certified
Containment currently certified, used and is used at all times
for all procedures that with agents being
generate aerosols handled.
Secondary Handwashing sink
Containment Locking doors Hands-free sink preferable
All surfaces can be Self-closing locked doors
disinfected Windows should not open
Windows have screens
and locks
Training/ Biosafety Training prior to starting experiments, including PI
Education PI must be appropriately educated/ trained or co-PI with someone who is
trained

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BSL-1 and BSL-2 Minimum Criteria

BSL-1 BSL-2 BSL-2+/ enhanced
Personal Protective No shorts/ tank tops Respiratory protection
Equipment and Lab Close-toed shoes Shoe covers
Attire Lab coat, safety glasses, gloves Bonnets
Long hair pulled back Disposable lab gowns
only
Equipment Dustpan/ brush/ tongs for glassware cleanup
EPA-Approved Disinfectant for lab surfaces
Mechanical pipets
Biohazard spill kit

Decontamination/ Waste must be Autoclave on-site required, waste must be

Disposal decontaminated prior decontaminated appropriately prior to
to disposal. disposal.

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BSL-1 and BSL-2 Minimum Criteria

BSL-1 BSL-2 BSL-2+/ enhanced
Signage Lab Sign possesses BSL-2 sign with entry/ exit procedures. Agent
biohazard symbol not posted unless in private hall.
Documentation SOPs as necessary Lab-Specific Biosafety Manual with all SOPs,
Sharps SOP when protocols, and training logs available in-lab
sharps present
Biosafety Biennial Annual
Inspections
Other No plants/ animals not part of experiments permitted
Integrated Pest Management Plan in place
Routine eye wash and chemical shower testing performed

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Windows with
screens and locks
Doors that
lock
Lab design
and
organization
BSL-1 Sink

Laboratory

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Biosafety Level 2 Laboratory

Sealed
Emergency Biological Safety Cabinet (BSC)
windows
shower and
eyewash station Non-porous work
surfaces

PPE: lab coat, safety

glasses, gloves
Autoclave Posted signage

Handwashing
sink
Self-closing
door w/ lock

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Biosafety Level 3 (BSL3)

• All of the criteria set for BSL-1 and BSL-2 are applicable in BSL-3.
• Level 3 (BSL-3/ BL3) laboratories have additional design and engineering
considerations.
• Personnel working in BSL-3 are required to have the appropriate
education, training, approvals to work in the laboratory.
• Separate training requirements in addition to facility design, practices,
procedures

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BSL-3 Laboratories

BSL-3 Biosafety Cabinet (used at

all times when manipulating agent)

Posted communication for lab

personnel (spills, equipment Autoclave in lab (best to have
issues, etc.) throughput to remove
decontaminated waste from
outside of lab)
Hands-free sink
Additional:
Directional airflow indicator Interlocking double door
Security- restricted access (keycard, entry with self-closing
Anteroom separation fingerprint scan, etc.) doors
Integral, cove molded
floors
Door sign with: biohazard symbol, Sealed windows
agents present (except when facing Lab-specific biosafety plan
public spaces), contact information, and in place
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Additional Considerations
• You lab may vary based on your work. The criteria provided by
the UWM Biosafety Program are the minimum criteria.
• Other requirements will be determined by the IBC.
• Plant biocontainment may require work in the segregated spaces
of the greenhouse.
• Animal facilities work with the Animal Care Program and have
specific requirements in addition to the standard requirements for
their respective BSLs.
• Teaching laboratories have unique considerations- personnel
training, lab layout, number of people working with agents at the
same time, etc.
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END

Module 3

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