Biological Laboratory Safety
REHS
Rutgers Environmental Health and Safety
REHS supports Rutgers University by providing
comprehensive and professional health, safety
and environmental services to the University
community.
Protect Rutgers employees, students and
visitors.
REHS ensures compliance with government
regulations
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Regulatory Agencies
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The safety standards and practices
described here have been developed
and are regulated by government
agencies including
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CDC (Center for Disease Control)
NIH (National Institutes of Health)
Agenda
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Lab Safety
Biohazards
Biosafety Levels
Biosafety Cabinets
Lab Safety
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Safe Laboratory
Practices
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Lab coats
Safety glasses
Proper footwear
Hair back
No food or drink in
the laboratory
Biohazard
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An agent of
biological origin that
can cause disease in
humans
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Microorganism
Toxin
Allergen
Biosafety
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The combined use of
laboratory practices,
laboratory facilities and
safety equipment
to work with potentially infectious
microorganisms.
Why use biosafety practices?
To protect:
 Workers/Students
 Products/Experimental results
 Environment/Laboratory classroom
Biosafety Levels
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BL-1: agents are not known to cause
disease
BL-2: agents are associated with human
disease
BL-3: agents are associated with human
disease and are potentially transmitted
as aerosols
BL-4: agents of life threatening nature
Biosafety Level 1 (BL-1)
Use BL-1 when working with:
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Well characterized agents
Agents that are not known to cause disease in health
humans
Agents that are of minimal hazard to lab personnel
and the environment
Examples of BL-1 Agents:
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E. coli JM109, DH5a
Saccharomyces cerevisiae
Biosafety Level 1 (BL-1)
Standard Work Practices
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Use mechanical pipetting devices
Wash hands frequently
Minimize splashes and aerosols
Decontaminate work surfaces daily
Handle wastes properly
Maintain insect and rodent control program
Biosafety Level 1 (BL-1)
Personal Protective
Equipment (PPE)
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Lab coat or apron
Safety glasses or goggles
Gloves as needed
Biosafety Level 2 (BL-2)
Use BL-2 practices when working with:
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Agents of moderate potential hazard to personnel
and the environment
Examples of BL-2 agents:
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Human blood or body fluids
E. coli 0157:H7
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Clostridium botulinum
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Retroviral vectors
Human cells in cell culture
Biosafety Level 2 (BL-2)
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Adequate illumination
Eyewash facility
Negative air pressure
Autoclave available
Biological safety cabinet
Lab must be separated from public
areas
Biosafety Level 2 (BL-2)
Standard Work Practices
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Use mechanical pipetting devices
Wash hands frequently
Minimize splashes and aerosols
Decontaminate work surfaces daily
Handle wastes properly
Maintain insect and rodent control program
Biosafety Level 2 (BL-2)
Special Practices
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Place used slides and coverslips in sharps
containers, never in any other receptacle.
Sharps containers are:
 Red in color
 Marked with the biohazard symbol
 Puncture resistant
 Leak proof
Biosafety Level 2 (BL-2)
Personal Protective Equipment (PPE)
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Lab coat or apron
Safety glasses or goggles
Gloves
Biosafety cabinet
 Aerosols or splashes
 Large volumes
 High concentrations
Biosafety Cabinets (BSCs)
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Provide product, personal and environmental
protection.
Various classes of BSCs are available
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Amount of air exhausted
Amount of employee protection
Biosafety Cabinets (BSCs)
Biosafety Cabinets (BSCs)
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Sterile air is exhausted from BSCs through a
HEPA filter.
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HEPA filter can trap particles to 0.3 microns
Chemical vapors and fumes pass through HEPA
filters
Biosafety Cabinets (BSCs)
Helpful Hints
 Enter straight into the cabinet, avoid
sweeping motions
 Don’t place materials on the grill
 Keep discard pan or bag inside the
cabinet
 Decon materials before removal from
the cabinet
Decontamination
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Sterilization: destroy all microbial life,
including spores
Disinfection: destroy a majority of
microbial life, but not necessarily spores
Antiseptic: germistat used on skin to
inhibit growth of microorganisms
Decontamination
Agent Selection
 Degree of killing required
 Nature of item to be treated
 Ease of use
 Safety
 Cost
 Type and amount of organism
Decontamination
Methods
 Heat: steam heat, dry heat, incineration
 Chemical: bleach, ethanol, hydrogen
peroxide, ethylene oxide,
paraformaldehyde
 Radiation
Decontamination
Disinfectants do not replace standard
microbiological practices and good
hygiene!
Biological Waste
Types
 Cultures, stocks
 Sharps
 Pipettes, tips and weigh boats
 Other materials
Biological Waste
BL-1 Waste
 Solids
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Collect and package in clear autoclave bags
Autoclave to sterilize
Dispose of in regular trash dumpster (as long as
no chemical or radioactive contaminants are
present)
Liquids
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Collect in containers with lids
Autoclave or treat with 10% bleach to sterilize
Dispose of down the drain (as long as no chemical
or radioactive contaminants are present)
Biological Waste
Sharps (needles, syringes, scalpel blades,
slides, blood vials, pasteur pipettes)
 Collect in approved sharps container
 Autoclave to sterilize
 Dispose of in medical waste boxes
Biosafety and REHS
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Protocol Approval
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REHS provisionally approves experimental
protocols.
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All protocols must be approved if they involve:
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recombinant DNA,
non-recombinant human, animal or plant pathogens,
human cell culture,
human tissue/blood experiments
Laboratory inspections
RU Biosafety Committee provides final approval to
protocols