The document discusses basic principles of biosafety. It begins by defining biosafety as safety from exposure to infectious agents. It then discusses biosafety levels 1-4 which provide increasing levels of containment for biological hazards. It also discusses important biosafety concepts like standard microbiological practices, safety equipment, facility design and construction that help prevent exposure to biological hazards in research and clinical settings.

1. Basic Principles of BiosafetyBasic Principles of Biosafety
Miguel Martin N. Moreno II, MDMiguel Martin N. Moreno II, MD
Scientist, SLMC, QC - RBDScientist, SLMC, QC - RBD
Founding Secretary, PhBBA NationalFounding Secretary, PhBBA National

2. What is Biosafety?
 Safety from exposure to
Infectious Agents
Smallpox

3. 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.

4. Biosafety in Various Disciplines
Biosafety 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. (Classified info.)

5. 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

6. 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

7. Biosafety Issues
 Laboratory Safety
 Bloodborne pathogens (BBP)
 Recombinant DNA (rDNA)
 Biological waste disposal
 Infectious substance and
diagnostic specimen shipping

8. 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

9. Biohazardous Materials
 Viruses
 Bacteria
 Fungi
 Chlamydiae/Rickettsiae
 Prions
 Recombinant DNA

10. Biohazardous materials
 Transgenic Plants, Animals and Insects

11. Biohazardous Materials
 Human and Primate Cells, Tissues, and
Body Fluids
 Brain Tissue from Demented Patients
 Viral Vectors
 Replication deficient viruses

12. Biosafety In Microbiological
and Biomedical Laboratories
BMBL
CDC/NIH Publication
Safety “Guidelines”
Regulations of Institution
receives
NIH funding
Code of Practice and “Gold”
Standard in Industry
Gold Standard
Clinical & Research Lab.
Lab. Animal Facilities
Biosafety Concepts
http://www.cdc.gov/od/ohs
/biosfty/bmbl5/bmbl5toc.h
tm

13. Canadian Laboratory Biosafety Guidelines
http://www.phac-
aspc.gc.ca/publicat/lbg-
ldmbl-
04/pdf/lbg_2004_e.pdf

14. •CEN Workshop Agreement (CWA15793)
•ftp://ftp.cenorm.be/public/CWAs/wokrshop31/CW
A15793.pdf
•WHO Laboratory Biosafety Manual
•http://www.who.int/csr/resources/publications/bios
afety/Biosafety7.pdf
•Sandia Laboratory Biosecurity Handbook
•http://www.crcpress.com/product/isbn/9780849364
754
Other Resources

15. Philippines
 Manual of Standards on National
Laboratory Biosafety & Biosecurity
 by the Task Force/TWG on the
Preparation of Standards on National
Laboratory Biosafety & Biosecurity as
per DP) No. 2006-2500
 DOH, National Center for Health Facility
Development, 17 March 2009

16. Biosafety Concepts
Principles of Biosafety
 Practice and Procedures
 Standard Practices
 Special Practices & Considerations
 Safety Equipment
 Facility Design and Construction
 Increasing levels of protection

17. 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

18. Biosafety Concepts
(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

19. (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

20. (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 Concepts

21. 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

22. Risk Group 1 Agents
 E.coli K-12
 Transgenic Plants
 Plasmids
 Fungi
 Mold
 Yeast

23. 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

24. Risk Group 2 Agents
 Human or Primate
Cells
 Herpes Simplex Virus
 Replication
Incompetent
Attenuated Human
Immunodeficiency
Virus
 Patient specimens

25. 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

26. BSL-2 Practices
 Limited access to lab
when work in progress
 Daily decontamination
 Mechanical pipetting
 Lab.coat, safety glasses
and gloves required
 Red bag & sharps
containers required

27. 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

28. Risk Group 3 Agents
 Human
Immunodeficiency
Virus
 Mycobacterium
tuberculosis
 Coxiella burnetii

29. Biosafety Level 3
Working 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

30. 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

31. 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”

32. 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

33. Biosafety Level-4
Working 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

34. Biosafety Level 4
 Lassa Fever Virus
 Ebola Hemmorrhagic
Fever Virus
 Marburg Virus
 Herpes B Virus

35. 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

36. 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

37. 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

38. Risk Assessment
In-Vitro In-Vivo Human Clinical
Trial

39. 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?

40. 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?

41. 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?

42. 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?

43. 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?

44. First Aid Measures
 Splash to Eye or Needlestick Injury
 Rinse thoroughly for 15 minutes at the eyewash or
sink
 Call Occupational Medicine
 Call Employee Health Services

45. Be safe, Be Biosafe, Be alive!
Maraming Salamat po!