Basic Biosafety Principles
EH&S Academy
Brenda J. Wong, UCSD Biosafety
Officer
October 2009
What is Biosafety?
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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.
Biosafety in Various Disciplines
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Biosafety is related to several fields
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ECOLOGY: referring to imported
life forms not indigenous to the
region (Reggie the alligator)
AGRICULTURE: reducing the
risk of alien viral or transgenic
genes, or prions such as
BSE/"MadCow“; reducing the risk
of food bacterial contamination
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MEDICINE: referring to
organs or tissues from biological
origin, or genetic therapy
products, virus; levels of lab
containment protocols BSL-1, 2,
3, 4 in rising order of danger
CHEMISTRY: i.e., nitrates in
water, PCB levels affecting
fertility
EXOBIOLOGY: i.e., NASA's
policy for containing alien
microbes that may exist on space
samples - sometimes called
"biosafety level 5"
Biosafety in Academic Research
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Research Universities:
Promoting safe laboratory
practices, and procedures;
proper use of containment
equipment and facilities;
provides advice on
laboratory design and risk
assesment of experiments
involving infectious agents,
rDNA in-vitro and in-vivo.
Bottom Line: Risk & Containment
Biohazard Symbol
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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
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Laboratory Safety
Bloodborne pathogens (BBP)
Recombinant DNA (rDNA)
Biological waste disposal
Infectious substance and
diagnostic specimen shipping
Biosafety Issues (con’t.)
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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
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Viruses
Bacteria
Fungi
Chlamydiae/Rickettsiae
Prions
Recombinant DNA
Biohazardous materials
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Transgenic Plants, Animals and Insects
Transgenic Insects
Biohazardous Materials
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Human and Primate Cells, Tissues, and
Body Fluids
Brain Tissue from Demented Patients
Viral Vectors
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Replication deficient viruses
Biosafety Concepts
Biosafety In Microbiological
and Biomedical Laboratories
“BMBL” (acronym)
CDC/NIH Publication
Safety “Guidelines”
Regulations of Institution receives
NIH funding
Code of Practice and “Gold”
Standard in Industry anl
Gold Standard
Clinical & Research Lab.
HHS Publication No. (CDC) 93-8395
The New BMBL
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Early print
edition….
Emphasis on “Risk
& Containment”
Biosafety Concepts
The BMBL
The BMBL continues
to be published by
the
CDC and the NIH
5th edition is now at
the printers
http://www.cdc.gov/od/ohs/bio
sfty/bmbl5/bmbl5toc.htm
Are the NIH Guidelines Optional?
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“Guidelines” does not mean
“optional”
They are a term and condition of
NIH funding for recombinant DNA
research.
From Kathryn Harris, NIH, OBA
Biosafety Concepts from
the BMBL
Principles of Biosafety
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Practice and Procedures
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Standard Practices
Special Practices & Considerations
Safety Equipment
Facility Design and Construction
Increasing levels of protection
Principles of Biosafety
Biosafety Levels 1-4 (BSL)
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Increasing levels of employee and environmental protection
Guidelines for working safely in research & medical
laboratory facilities
Animal Biosafety Levels 1- 4
(ABSL)
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Laboratory animal facilities
Animal models that support research
Guidelines for working safely in animal research facilities
Biosafety Concepts
The BMBL
(1) Standard Microbiological Practices
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Most important concept / Strict adherence
Aware of potential hazard
Trained & proficient in techniques
Supervisors responsible for:
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Appropriate Laboratory facilities
Personnel & Training
Special practices & precautions
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Occupational Health Programs
Biosafety Issues
The BMBL
(2) Safety Equipment
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Primary Containment Barrier
Minimize exposure to hazard
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Engineering controls/ equipment
Personal Protective Equipment (PPE)
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Prevent contact / Contain aerosols
Gloves, gowns, Respirator, Face shield, Booties
Biological Safety Cabinets
Covered or ventilated animal cage systems
Biosafety Concepts
The BMBL
(3) Facility Design and
Construction
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Secondary Barrier/ Engineering
controls
Contributes to worker protection
Protects outside the laboratory
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Environment & Neighborhood
Ex. Building & Lab design,
Ventilation, Autoclaves, Cage wash
facilities, etc.
Laboratory Design
“Warehouse Type Lab”
Discussion
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What are some of the
negatives and
positives of this open
lab concept?
Biosafety Level-1
Concepts of Biosafety
Biosafety Level-1 (BSL-1 or ABSL-1)
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Well characterized agents
Agents not known to cause disease (in healthy human
adults; now healthy immunocompetent adults)
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Prophylactic treatment available
Open bench procedures
Animals in open cage system or open environment
(outdoors)
Good laboratory practices
Risk Group 1 Agents
E.coli K-12
 Transgenic Plants
 Plasmids
 Fungi
 Mold
 Yeast
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BSL-1 Practices
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Bench-top work allowed
Daily Decontamination
Manual pipetting
Required Handwashing
Red bag waste
Bio cabinet not required
(unless creating aerosols)
2˚ containment
Risk Group 2 Agents
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Human or Primate
Cells
Herpes Simplex Virus
Replication
Incompetent
Attenuated Human
Immunodeficiency
Virus
Patient specimens
BSL-2 Practices
Concepts of Biosafety
Practices & Procedures
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Agents associated w/ human disease
Treatment for disease available
Agent poses moderate hazard to personnel and
environment
Direct contact or exposure
Percutaneous exposure
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Scratch, Puncture, Needle stick
Mucus membrane exposure
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Eyes, Mouth, open cut
BSL-2 Practices
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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)
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Biohaz. Sign posted at
entrance to lab
Label all equipment
(incubators, freezers, etc.)
TC room – negative air flow
Documented training
Baseline serology or prevaccination may be required
Risk Group 3 Agents
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Human
Immunodeficiency
Virus
Mycobacterium
tuberculosis
Coxiella burnetii
Biosafety Level 3
Working in High Containment
Biosafety Level-3 (BSL-3 or
ABSL-3)
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Indigenous or exotic agents
Aerosol transmission
Serious health effects
Treatment may or may not exist
BSL-3 Practices
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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
No sharps unless absolutely necessary
BSL-3 Practices (con’t)
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Aerosol minimization procedures required
Wrap around disposable clothing is
required. Specialized equipment may be
required depending upon procedures
Biohaz. Signs and labels posted
Air flow from low hazard to high hazard
“Pressure Mapping”
BSL-3 Practices (con’t)
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Bench top work not permitted
Documented training and personnel competency
certification (for BSL-3 procedures)
Baseline serology
Spills – report immediately and treat accordingly
Vaccinations/post exposure protocols and SOP’s,
Biosafety Manual, Biosafety Officer
UCSD’s BSL-3
Biosafety Level-4
Working in High Containment
Biosafety Level-4
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Builds on BSL-3/ ABSL-3 practices
Maximum containment facilities
Pressurized Containment Suite
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BSL-3 + Class III Biosafety Cabinet
Chemical decontamination showers
Liquid effluent collection / decontamination
No BSL-4 labs exist at UCSD
Biosafety Level 4
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Lassa Fever Virus
Ebola Hemmorrhagic
Fever Virus
Marburg Virus
Herpes B Virus
Biosafety Concepts
Working in High Containment
Biosafety Level-4 (BSL-4 or ABSL-4)
Dangerous/exotic agents
 Life threatening disease
 Aerosol transmission
 Agents of unknown risk
of transmission or health affects
 No known treatment
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Animal Biosafety Level-4
Working in High Containment
General Good Lab Technique
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Hygienic Practices
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No Smoking, Eating, Applying cosmetics, lip
balm, contacts
Wash hands after procedures
Decontaminate lab bench before and after
work
General Operational Practices
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Proper attire
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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/msdsftss/index.html
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Animal Containment Points
CDC - 1957
CDC & UCSD - 2005
Courtesy of Paul Vinson, CDC
Discussion # 2
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Based on what you know about Biosafety
Levels, Practices and Operational Controls,
what are some discussion issues for
conducting Biohazard risk assessments?
How do you approach risks when
addressing a particular organism?
Risk Assessment
In-Vitro
In-Vivo
Human Clinical
Trial
Addressing Risk Assessments
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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
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Are personnel trained? Do
personnel understand the
organism, infectious dose and
symptoms?
What are their experimental
procedures?
Will they be transporting the
material? Shipping intra, interstate or international?
Are they doing tissue culture?
Do they have adequate
containment equipment?
Tom Pugh
Risk Assessment, Con’t
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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
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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
 Don PPE; 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
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Splash to Eye or Needlestick Injury
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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?
Resources
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UCSD Biosafety: http://blink.ucsd.edu/safety/researchlab/biosafety/
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Biological Safety MSDS: http://www.phacaspc.gc.ca/msds-ftss/index-eng.php
NIH BMBL:
http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
See Handout for additional resources..
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Questions?
Brenda Wong
Bwong@ucsd.edu
858 534-6059
Cell: 858 583-3277
Additional Resources:
Biosafety Resources.doc