The document provides an introduction to biosafety, explaining that it aims to reduce risk of exposure to infectious materials through proper safety precautions and procedures. It discusses the need for biosafety in laboratories processing infectious agents and around recombinant DNA to protect workers and the environment. The document also outlines different biosafety levels and associated practices, containment facilities, risk groups of pathogens, and considerations for risk assessments.

1. Introduction to Biosafety

2. Biosafety
According to the Centre Of Disease Control and
Prevention (CDC)
“ Biosafety is the application of safety
precautions that reduce a laboratorians risk of
exposure to a potentially infectious material
and limit contamination of the work
environment and ultimately the community.”

3. Why biosafety???

4. Need of biosafety
Lab has hazards of processing
infectious agents
Accidental threat to workers
and environment
To have adherence with safety
regulations while dealing with
highly infectious agents

5. Scope of biosafety
Biosafety
Ecology Agriculture Medicine Exobiology Chemistry

6. Biosafety In Academic Research
• Promoting safe laboratory practices, and
procedures
• proper use of containment equipment and
facilities
• provides advice on laboratory design and risk
assessment of experiments involving
infectious agents, rDNA in-vitro and in-vivo.

7. Biohazard Symbol
• Charles Baldwin at
National Cancer
Institute at NIH.
• Symbol to be
“memorable but
meaningless” so it could
be learned.

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

9. Biosafety Issues
• 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

10. Biohazardous Materials
• Viruses
• Bacteria
• Fungi
• Chlamydiae/Rickettsiae
• Prions
• Recombinant DNA

11. Biohazardous Materials
• Transgenic Plants, Animals and Insects

12. Basis for the Classification of Biohazardous Agents
by Risk Group
Risk Group 1 (RG-1)
Agents that are not associated with disease in
healthy adult humans.
Risk Group 2 (RG-2)
Agents that are associated with human disease
which are rarely serious and for which
preventive or therapeutic interventions are
often available.
Risk Group 3 (RG-3)
Agents that are associated with serious or
lethal human disease for which preventive or
therapeutic interventions may be available
(high individual risk but low community risk).
Risk Group 4 (RG-4)
Agents that are likely to cause serious or lethal
human disease for which preventive or
therapeutic interventions are not usually
available (high individual risk and high
community risk)

13. Risk group classification

14. Risk Assessment
• Pathogenicity…
• Virulence - the severity of disease..
• Transmission route…
• Agent stability - survival in environment or
otherwise prolonged viability (spore formation).
• Infectious dose..
• Antibiotic resistance…
Risk Assessment

15. Biosafety In Microbiological
and Biomedical Laboratories
“BMBL” (acronym)
CDC/NIH Publication
Safety “Guidelines”
Regulations of Institution receives
NIH funding
Clinical & Research Lab.
Lab. Animal Facilities
Biosafety Concepts
HHS Publication No. (CDC) 93-8395

16. The New BMBL
• Early print edition….
• Emphasis on “Risk
& Containment”

17. The BMBL continues
to be published by
the
CDC and the NIH
5th edition is now at
the printers
Biosafety Concepts
The BMBL

18. Are the NIH Guidelines Optional?
 “Guidelines” does not mean
“optional”
 They are a term and condition of
NIH funding for recombinant DNA
research.

19. 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

20. 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

21. 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

22. 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

23. 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.

25. Laboratory Design
“Warehouse Type Lab”

26. Discussion
• What are some of the
negatives and
positives of this open
lab concept?

27. 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)
• Prophylactic treatment available(disease prevention)
• Open bench procedures
• Animals in open cage system or open
environment (outdoors)
• Good laboratory practices

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

29. 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

30. 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

31. 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

32. 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

33. Risk Group 3 Agents
• Human
Immunodeficiency
Virus
• Mycobacterium
tuberculosis
• Coxiella burnetii

34. 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

35. 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

36. 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”

37. 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

38. UCSD’s BSL-3

39. 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
• No BSL-4 labs exist at UCSD

40. Biosafety Level 4
• Lassa Fever Virus
• Ebola Hemorrhagic
Fever Virus
• Herpes B Virus

41. 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

42. Animal Biosafety Level-4
Working in High Containment

43. 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

44. 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

45. Animal Containment Points
CDC - 1957
CDC & UCSD - 2005
Courtesy of Paul Vinson, CDC

46. 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?

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

49. 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?

50. 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

51. 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?

52. 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?

53. 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?

54. First Aid Measures
• Splash to Eye or Needlestick Injury
– Rinse thoroughly for 15 minutes at the eyewash or sink
– Call Occupational Medicine  …………