Basic Biosafety Principles

What is Biosafety?
◼ Safety from exposure to
Infectious Agents

Smallpox
 What is Biosafety?

Sunday, Sep. 20, 2009

Did the Plague Kill Illinois Scientist?
By AP
(AP / CHICAGO) — The University of Chicago Medical Center says the infection that killed a scientist
may be connected to bacteria he researched that causes the plague.
The university said Saturday that its researcher studied the genetics of harmful bacteria including
Yersinia pestis, which causes the illness. He died Sept. 13. His name and age haven't been
released
The medical center says the bacteria he worked with was a weakened strain that isn't known to cause
illness in healthy adults. The strain was approved by the Centers for Disease Control and
Prevention for laboratory studies.
An autopsy found no obvious cause of death but did find the presence of the bacteria. More tests are
planned. No other illnesses have been reported.
Biohazard Symbol
◼ Charles Baldwin at
National Cancer
Institute at NIH.
◼ Symbol to be
“memorable but
meaningless” so it
could be learned.
◼ Blaze orange – most
visible under harsh
conditions
 Biosafety Issues
◼ Laboratory Safety
◼ Bloodborne pathogens (BBP)
◼ Recombinant DNA (rDNA)
◼ Biological waste disposal
◼ Infectious substance and
diagnostic specimen shipping
 Biosafety Issues (con’t.)
◼ Respiratory Protection
◼ Bioterrorism and Select agents
◼ Mold and indoor air quality
◼ Occupational safety and health in the use of
research animals
◼ Biohazards used in animal models
Biohazardous Materials
◼ Viruses
◼ Bacteria
◼ Fungi
◼ Chlamydiae/Rickettsiae
◼ Recombinant DNA
Biohazardous materials
◼ Transgenic Plants, Animals and Insects
 Biohazardous Materials

◼ Human and Primate Cells, Tissues, and

Body Fluids
◼ Brain Tissue from Demented Patients
◼ Viral Vectors
◼ Replication deficient viruses
 Biosafety Concepts
The BMBL
(1) Standard Microbiological Practices
◼ Most important concept / Strict adherence
◼ Aware of potential hazard
◼ Trained & proficient in techniques
◼ Supervisors responsible for:
◼ Appropriate Laboratory facilities
◼ Personnel & Training
◼ Special practices & precautions
◼ Occupational Health Programs
 Biosafety Issues
The BMBL
(2) Safety Equipment
◼ Primary Containment Barrier
◼ Minimize exposure to hazard
◼ Prevent contact / Contain aerosols
◼ Engineering controls/ equipment
◼ Personal Protective Equipment (PPE)
◼ Gloves, gowns, Respirator, Face shield, Booties
◼ Biological Safety Cabinets
◼ Covered or ventilated animal cage systems
 Biosafety Concepts
The BMBL
(3) Facility Design and
Construction
◼ Secondary Barrier/ Engineering
controls
◼ Contributes to worker protection
◼ Protects outside the laboratory
◼ Environment & Neighborhood
◼ Ex. Building & Lab design,
Ventilation, Autoclaves, Cage wash
facilities, etc.
 Biosafety Level-1
Concepts of Biosafety
Biosafety Level-1 (BSL-1 or ABSL-1)
◼ Well characterized agents
◼ Agents not known to cause disease (in healthy human
adults; now healthy immunocompetent adults)
 Risk Group 1 Agents
◼ E.coli K-12
◼ Transgenic Plants
◼ Plasmids
◼ Fungi
◼ Mold
◼ Yeast
BSL-1 Practices
◼ Bench-top work allowed
◼ Daily Decontamination
◼ Manual pipetting
◼ Required Handwashing
◼ Red bag waste
◼ Bio cabinet not required
(unless creating aerosols)
 Risk Group 2
Pathogenic for humans
Unlikely a serious hazard
Treatment and preventive
measures available
Limited risk of spread of
infection

CDC, Yersinia pestis laboratory

 Risk Group 2 Agents
◼ Human or Primate
Cells
◼ Herpes Simplex Virus
◼ Replication
Incompetent
Attenuated Human
Immunodeficiency
Virus
◼ Patient specimens
BSL-2 Practices
◼ Limited access to lab
when work in progress
◼ Daily decontamination
◼ Mechanical pipetting
◼ Labcoat, safety glasses
and gloves required
◼ Red bag & sharps
containers required
 BSL-2 Practices (con’t)
◼ Biohaz. Sign posted at
entrance to lab
◼ Label all equipment
(incubators, freezers, etc.)
◼ Documented training
 Risk Group 3
Pathogenic, cause serious disease
Effective treatment and preventive
measures usually available
Little person-to-person spread

Laboratory in Lyon France

 Risk Group 3 Agents
◼ Human
Immunodeficiency
Virus
◼ Mycobacterium
tuberculosis
◼ Coxiella burnetii
 BSL-3 Practices
◼ Public access NOT permitted
◼ Daily decontamination after spill and
upon completion of experiment
◼ Autoclave required and waste is
disposed at the end of day
◼ Required foot activated handwashing
sink and controls
BSL-3 Practices (con’t)
◼ Wrap around disposable clothing is
required. Specialized equipment may be
required depending upon procedures
◼ Biohaz. Signs and labels posted
BSL-3 Practices (con’t)
◼ Bench top work not permitted
◼ Documented training and personnel competency
certification (for BSL-3 procedures)
◼ Spills – report immediately and treat accordingly
◼ Vaccinations/post exposure protocols and SOP’s,
Biosafety Manual, Biosafety Officer
UCSD’s BSL-3
 Risk Group 4
Lethal, pathogenic agent
Readily transmittable
◼ direct, indirect
Effective treatment and
preventive measures not usually
available

National Institute for Infectious

Diseases, Rome, Italy
Biosafety Level 4
◼ Lassa Fever Virus
◼ Ebola Hemmorrhagic
Fever Virus
◼ Marburg Virus
◼ Herpes B Virus
Animal Biosafety Level-4
Working in High Containment
General Good Lab Technique
◼ Hygienic Practices
◼ No Smoking, Eating, Applying cosmetics, lip
balm, contacts
◼ Wash hands after procedures
◼ Decontaminate lab bench before and after
work
 General Operational Practices
◼ Proper attire
◼ Minimum – lab coat, safety glasses, gloves
◼ Plan your work
◼ Know in advance what you are working with
◼ Read available resources (MSDS)
http://www.hc-sc.gc.ca/pphb-dgspsp/msds-
ftss/index.html
Animal Containment Points

CDC - 1957
CDC & UCSD - 2005

Courtesy of Paul Vinson, CDC

 Risk Assessment

In-Vitro In-Vivo Human Clinical

Trial
 Addressing Risk Assessments
◼ What is the organism?
◼ Is it Wild-type, attenuated, irradiated, or
chemically treated? Look at kill data or kill
curves.
◼ What is the max. concentration, volume,
infectious dose?
◼ What is the work space like?
◼ Aerosolizing procedures? How do they contain
their aerosols?
Risk Assessment, con’t
◼ Are personnel trained? Do
personnel understand the
organism, infectious dose and
symptoms?
◼ What are their experimental Tom Pugh
procedures?
◼ Will they be transporting the
material? Shipping intra, inter-
state or international?
◼ Are they doing tissue culture?
◼ Do they have adequate
containment equipment?
Risk Assessment, Con’t
◼ Are they doing this
work in-vivo? Have
you consulted and
discussed this with
the Vets and IACUC
to determine special
needs and housing?
◼ Waste issues
addressed?
◼ Pregnancy issues with
the organisms?
Risk Assessment, con’t
◼ Do they share their
Tissue Culture room?
◼ Do they have more than
1 Biosafety Cabinet?
◼ Occupational Health
informed and set up to
receive patient or offer
counseling?
 Accidental Spills

▪ Evacuate area, alert personnel and

cordon off so that aerosols may settle
▪ Cover with paper towels and apply
bleach (1 part bleach : 9 parts water
▪ Allow 15 – 20 min contact time
▪ Wipe up working towards center
▪ Use tongs if broken glass is involved
▪Is Recombinant DNA involved?
First Aid Measures

◼ Splash to Eye or Needlestick Injury

◼ Rinse thoroughly for 15 minutes at the eyewash or
sink
◼ Call Occupational Medicine → 619 471-9210
◼ Call EH&S to report exposure – 858 534-5366
What Helps?