BioSafety Training
Contact Information
• Research & Development
http://www.pvamu.edu/pages/108.asp
– Institutional Review Board (IRB)
– Institutional Animal Care and Use Committee
(IACUC)
– Institutional Biosafety Committee (IBC)
• Compliance Officer: Dr. M Shelton
New Employees
• Should receive proper
training prior to
beginning work
• Should be supervised
when working with
biological materials
until they can
demonstrate proper
techniques and
procedures
Aseptic Techniques and Procedures
• Prior to manipulation
– Use sterile components
– Use engineered controls when appropriate (i.e. BSC)
– Ensure components are present in BSC
• During manipulation
– Open vessel for minimum time required
– Avoid cross contamination (e.g., using a loop that has
contacted the outside of the vessel)
Aseptic Techniques and
Procedures
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Use Universal Precaution
No mouth pipetting
No food, drink, tobacco products
Do not apply cosmetics or remove contacts
Work surfaces should be decontaminated every day and
after any spill
• Wash hands:
– After working with a biohazard
– After removing PPE
– Before leaving lab
• Do not touch face while working with biological material
• Do not wear PPE outside the lab
Universal Precautions
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Assume that all samples are infectious
Use good laboratory practices
Use protective barriers
Wash hands and exposed skin
Avoid skin punctures
Avoid contact with skin openings
Immunizations
Biological Safety Levels
• BSL-1
– General Biology
• BSL-2
– Pathogens
(not airborne)
• BSL-3
– Highly Infectious or
Airborne
• BSL-4
– Extreme Hazards (full
containment)
*5th Edition is on-line
Biological Waste Disposal
Liquids:
Sewer
Solids:
Decontaminate
Autoclave
Autoclave
Animals:
Incinerate
Bio-digest
Sharps:
Encapsulate
or
Trash
Trash
Choosing a Disinfectant
• When choosing a
disinfectant
– Consider the organism
– Characteristics of the
work area
• Frequently disinfect
surfaces and
equipment
Types of Disinfectants
Alcohols
Ethyl or Isopropyl Alcohol at 70-80% concentration is a good
general purpose disinfectant; not effective against bacterial
spores
Phenols
Effective against vegetative bacteria, fungi, and viruses
containing lipids; unpleasant odor
Formaldehyde
Concentration of 5-8% formalin is a good disinfectant against
vegetative bacteria, spores, and viruses; known carcinogen;
irritating odor
Quaternary
Ammonium
Compounds
Cationic detergents are strongly surface active; extremely
effective against lipoviruses; ineffective against bacterial
spores; may be neutralized by anionic detergents (i.e. soaps)
Chlorine
Low concentrations (50-500ppm) are active against vegetative
bacteria and most viruses;
Higher concentrations (2500ppm) are required for bacterial
spores; corrosive to metals, must be prepared fresh
Iodine
Recommended for general use; effective against vegetative
bacteria and viruses; less effective against bacterial spores
Wet Heat Sterilization
• Occurs when
contaminate reaches
– 15 psi
– 250F
– 30 min
• Potential Problems
– Dense Loads
– Poor Heat Conductors (e.g.
plastics)
– Containers preventing
steam penetration (some
plastic bags and/or double
bagging)
Autoclave
• Testing Loads
– Autoclave Tape
• Verifies materials have
been in autoclave
– Biological Indicator
• Most Effective
• Test should be
conducted as normally
used
Autoclave (loading)
• Do not double bag or tightly
seal
• Do not place sharp objects in
bag
• Shallow metal pans are more
effective than tall plastic tubs
• Vessels with liquid should not
be capped
• Add water to dry waste bags
Autoclave (removing waste)
• Ensure autoclave is “off” before
opening
• Ensure steam pressure is down
• Open door (no more than .5 inch)
slowly keeping head and hands
from opening
• Wait at least 10 minutes (or
manufacturers recommendation)
before removing anything
• Wear heat resistant gloves
• Clean any spill immediately
• Take bag to dumpster
Biological Waste Disposal
• For solid waste – deface
biological symbol on
autoclave bag (required),
place bag in black trash
bag (recommended), and
place in dumpster
• Biohazard bags are only
to be used for
biohazardous waste!
• For all biological waste –
trained lab personnel
need to dispose of waste
(custodial staff is not
responsible)
Encapsulation
• Metal Sharps
– Decontaminate
– Use rigid container
– Encapsulate (plaster
of paris)
– Label “Encapsulated
sharps”
– Place in dumpster
• Glass Sharps
– Decontaminate
– Seal in puncture
resistant container
– Label “Noncontaminated
glassware”
– Place in dumpster
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Biological Safety Cabinet
• Airflow + HEPA
filtration
• Keep Clean
• Set-up Before use
with BioHazards
• Inspected/Certified
– Initially
– After relocation or
repair
– Annually or
semiannually
Working in a Biological Safety
Cabinet
• Preparation:
– Turn off UV light
– Wipe work surface with
appropriate disinfectant
– Place only needed items in
cabinet
– Segregate items for clean
and dirty materials
– Provide waste container
inside cabinet
– Restrict entry into lab and
movement in lab
Working in a Biological Safety
Cabinet
• Use:
– Wear lab coat and gloves
– Keep work at least 4” from
glass view panel
– Limit arm movement
– Limit use of burner
– Place disinfectant-soaked
towel on the work surface
to contain spill
– Control tissue and loose
paper
Working in a Biological Safety
Cabinet
• Completion:
– Decontaminate surface or
enclose items which came
in contact with biohazard
– Cover waste container
– Allow cabinet to operate
15-20 min without activity
or leave it on
Biological Spills
• Develop a Plan of Action
prior to working with
biological materials
• Immediate actions:
– warn others
– leave the room; close the
door
– remove contaminated
garments
– wash hands
– notify supervisor
– seek medical attention, if
necessary
Biological Spill
• Clean Up Procedure
– wait for aerosols to
settle
– put on protective
clothing
– cover with towels;
allow contact time
– apply disinfectant
– wipe up; mop floor
– autoclave wastes
Records
• Metal Sharps – date of
treatment, quantity,
method, and name of
person (maintain 3 years)
• Autoclave –
– date of treatment,
quantity, method,
name of person
– date of verification test
and results
Transporting Infectious Materials
• On Campus
– Use leak-proof container
– Place primary container into sealed leak-proof
secondary container
– Use rigid transport container
– Label transport container with biohazard symbol,
material, emergency contacts
– Secure transport container in campus vehicle (DO NOT
use your personnel vehicle for transportation)
Transporting Infectious
Materials
• Off campus
– Follow federal and
international
regulations
– Only by trained
employees or contact
EHS @ 1745 / 1746