Download Biological Laboratory Safety

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