Basic Biosafety Principles EH&S Academy Brenda J. Wong, UCSD Biosafety Officer October 2009.
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Transcript of Basic Biosafety Principles EH&S Academy Brenda J. Wong, UCSD Biosafety Officer October 2009.
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Basic Biosafety Principles
EH&S Academy
Brenda J. Wong, UCSD Biosafety Officer
October 2009
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What is Biosafety?
Safety from exposure to Infectious Agents
Smallpox
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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.
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Biosafety in Various DisciplinesBiosafety is related to several fields
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
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"
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Biosafety in Academic Research 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
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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
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Biosafety Issues Laboratory Safety Bloodborne pathogens (BBP) Recombinant DNA (rDNA) Biological waste disposal Infectious substance and diagnostic specimen shipping
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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
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Biohazardous Materials Viruses Bacteria Fungi Chlamydiae/Rickettsiae Prions Recombinant DNA
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Biohazardous materials Transgenic Plants, Animals and Insects
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Transgenic Insects
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Biohazardous Materials
Human and Primate Cells, Tissues, and Body Fluids
Brain Tissue from Demented Patients Viral Vectors
Replication deficient viruses
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Biosafety In Microbiologicaland Biomedical Laboratories“BMBL” (acronym)
CDC/NIH PublicationSafety “Guidelines”Regulations of Institution receives NIH funding
Code of Practice and “Gold” Standard in Industry anlGold Standard
Clinical & Research Lab.Lab. Animal Facilities
Biosafety Concepts
HHS Publication No. (CDC) 93-8395
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The New BMBL
Early print edition….
Emphasis on “Risk
& Containment”
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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/biosfty/bmbl5/bmbl5toc.htm
Biosafety ConceptsThe BMBL
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Are the NIH Guidelines Optional?
“Guidelines” does not mean “optional”
They are a term and condition of NIH funding for recombinant DNA research.
From Kathryn Harris, NIH, OBA
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Biosafety Concepts from the BMBL
Principles of Biosafety Practice and Procedures
Standard Practices Special Practices & Considerations
Safety Equipment Facility Design and Construction Increasing levels of protection
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Principles of Biosafety
Biosafety Levels 1-4 (BSL) Increasing levels of employee and environmental protection Guidelines for working safely in research & medical
laboratory facilities
Animal Biosafety Levels 1- 4 (ABSL)
Laboratory animal facilities Animal models that support research Guidelines for working safely in animal research facilities
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Biosafety ConceptsThe 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
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Biosafety IssuesThe 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
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Biosafety ConceptsThe 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.
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Laboratory Design
“Warehouse Type Lab”
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Discussion
What are some of the negatives and positives of this open lab concept?
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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)
Prophylactic treatment available Open bench procedures Animals in open cage system or open environment
(outdoors) Good laboratory practices
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Risk Group 1 Agents E.coli K-12 Transgenic Plants Plasmids Fungi Mold Yeast
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BSL-1 Practices
Bench-top work allowed Daily Decontamination Manual pipetting Required Handwashing Red bag waste Bio cabinet not required
(unless creating aerosols) 2˚ containment
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Risk Group 2 Agents Human or Primate
Cells Herpes Simplex Virus Replication
Incompetent Attenuated Human Immunodeficiency Virus
Patient specimens
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BSL-2 Practices Concepts of Biosafety
Practices & Procedures Agents associated w/ human disease Treatment for disease available Agent poses moderate hazard to personnel and
environment Direct contact or exposure Percutaneous exposure
Scratch, Puncture, Needle stick Mucus membrane exposure
Eyes, Mouth, open cut
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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
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BSL-2 Practices (con’t)
Biohaz. Sign posted at entrance to lab
Label all equipment (incubators, freezers, etc.)
TC room – negative air flow Documented training Baseline serology or pre-
vaccination may be required
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Risk Group 3 Agents Human
Immunodeficiency Virus
Mycobacterium tuberculosis
Coxiella burnetii
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Biosafety Level 3Working in High Containment
Biosafety Level-3 (BSL-3 or ABSL-3)
Indigenous or exotic agents Aerosol transmission Serious health effects Treatment may or may not exist
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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 No sharps unless absolutely necessary
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BSL-3 Practices (con’t)
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”
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BSL-3 Practices (con’t)
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
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UCSD’s BSL-3
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Biosafety Level-4Working in High Containment
Biosafety Level-4 Builds on BSL-3/ ABSL-3 practices Maximum containment facilities Pressurized Containment Suite
BSL-3 + Class III Biosafety Cabinet Chemical decontamination showers Liquid effluent collection / decontamination No BSL-4 labs exist at UCSD
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Biosafety Level 4 Lassa Fever Virus Ebola Hemmorrhagic
Fever Virus Marburg Virus Herpes B Virus
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Biosafety ConceptsWorking 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-4Working in High Containment
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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
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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
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Animal Containment Points
CDC - 1957CDC & UCSD - 2005
Courtesy of Paul Vinson, CDC
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Discussion # 2 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?
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Risk Assessment
In-Vitro In-Vivo Human ClinicalTrial
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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?
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Risk Assessment, con’t 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, inter-state or international?
Are they doing tissue culture? Do they have adequate
containment equipment?
Tom Pugh
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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?
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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?
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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 involvedIs Recombinant DNA involved?
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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
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What Helps?
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Resources UCSD Biosafety: http://blink.ucsd.edu/safety/research-
lab/biosafety/
Biological Safety MSDS: http://www.phac-aspc.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
858 534-6059
Cell: 858 583-3277
Additional Resources:
Biosafety Resources.doc
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