1. Biosafety in Research Laboratories

2. Definitions Biohazard
An agent of biological origin that has the capacity to produce negative effects on humans, plants or animals.
Biosafety
The application of: lab practices and procedures, specific lab facilities and safety equipment to protect against exposure to potentially infectious material

3. What is Biohazardous Material?
Bacteria
Viruses
Parasites
Fungi
Human and Non-Human Primate Material
Recombinant DNA
Animals
Biological Toxins

4. How Can You Be Exposed?
Inhalation -80% of lab acquired infections (LAI)
Perform experiments in biological safety cabinets
Use respiratory protection
Remaining 20% LAI are a combination of:
Percutaneous
Avoid sharps when possible, use plastic pipettes and disposable glassware
Mucous Membrane (eyes, nose and mouth)
Wash hands often, avoid touching face
Ingestion
Wash hands often, No eating, drinking, smoking or applying cosmetics
All exposures must be reported to the PI, Occ. Health, Biosafety

5. How Can You Protect Yourself?
Administrative Controls
Use attenuated or non-virulent strains, require restraint during animal injection, avoid sharps and glassware, require decontamination, Standard Operating Procedures, training
Engineering Controls
Biosafety Cabinets, Sealed Rotors/Safety Cups, Pipette Aids, Safety Enclosures for Cell Sorters
Personal Protective Equipment
Lab Coat / gown, gloves, goggles, facial protection
Close toed shoes

6. Guidelines and Regulations
                              
Guidelines and Regulations
Biosafety in Microbiological and Biomedical Laboratories (CDC)
NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH)
OSHA Bloodborne Pathogens Standard (OSHA)
Committee on Microbiological Safety (COMS)

7. Why Biosafety Guidelines?
Protection
Self, Co-workers, Lab support personnel
Emergency Responders
Experiment and Materials
Environment
Compliance with NIH, CDC, OSHA, City of Boston, and Town of Belmont

8. Safety Equipment (Primary Barriers)
Biosafety cabinets (BSCs)
Safety centrifuge cups and rotors
Pipetting Devices
Vacuum line HEPA Filters
Personal protective clothing
Gloves (remember the one glove rule)
Gowns/Lab coats (should remain inside the lab)
Eye and face protection
Close toed shoes
Graphics Ideas:
Back of BMBL has line drawings (p ) of BSC
Picture or drawing of Eye Protection
Centrifuge Cups
JYR: For the intro, since these are additive, highlight gloves, gowns and pipetting devices.
(I omitted >> Safety centrifuge cups and rotors << due to space limitations. Could add if necessary.)

9. Biological Safety Cabinets
Must be Certified Annually
Air Systems Technologies (508)
BSC Provide:
Product Protection
Personnel Protection
Environmental Protection
Use biosafety cabinets (class II) for work with infectious agents involving:
Aerosols and splashes
Large volumes, High concentrations
BL2 materials

10. Biological Safety Cabinets
Protection against particulates only
No gas or vapor protection
Air flow is easily disrupted
Don’t cover the grill
Move arms slowly in and out of cabinet
Walk slowly
70% return 30% exhausted

11. Centrifuges Safety Cups and Rotors
Load and unload safety cups inside the BSC

12. Vacuum Line HEPA Filter Protection
Placed in between the vacuum system and the aspiration flasks

13. Biosafety Levels
BSL-1: agents not known to cause disease (in healthy adults)
BSL-2: agents associated with disease
BSL-3: indigenous/exotic agents associated with human disease and with potential for aerosol transmission
BSL-4: dangerous/exotic agents of life threatening nature

14. Biosafety Levels BSL-2S (with Stipulation)
Harvard classification not recognized nationally
Work can be done inside a BL2 structural facility while practicing certain BL3 practices and procedures
disposable sleeve covers or gowns,.....
Stipulated by the Institutional Biosafety Committee (COMS)

15. Biosafety Level 1 Immunocompromised workers are at risk
Standard Operating Procedures
Minimize splashes and aerosols
PPE
Decontaminate work surfaces daily
Decontaminate wastes
Maintain insect & rodent control program
Examples:
Adeno associated virus
Murine leukemia virus
Non pathogenic E. coli
Animal tissue and cells (other than NHP)
Biosafety Level 1

16. Biosafety Level 1 Laboratory Facilities (Secondary Barriers)

17. Biosafety Level 2 • Infectious dose will vary based on:
Agent
Exposure time
Exposure route
• Immunization or antibiotic treatment may be available
Extreme precaution with contaminated needles or sharp instruments
• Examples:
Hepatitis B Virus
Influenza
Salmonella

18. Biosafety Level 2 Biosafety Level 2 Signs
Policies and procedures for entry
Biohazard warning signs
Specific hazard training with annual updates
Graphic of sign for entrance to lab with
Biohazard symbol
BSL-2
Agent:
PPE:
Contact:

19. Biosafety Level 2 Laboratory Facilities (Secondary Barriers)
Hand washing sink located near laboratory exit
Autoclave available
Eyewash/Shower station available
Biosafety Cabinets
Autoclave picture here

20. Biosafety Level 2 Laboratory Facilities (Secondary Barriers)

21. Special Practices Use leak-proof transport containers Graphics
Leak proof transport containers (Pic of Nalgene container)

22. Biological Spill Kits Location Contents Spill Procedures
Inside Tissue Culture Rooms
Contents
Gloves, Goggles/shield, N95 Respirator, Lab coat/gown
Absorbent material
Bleach or suitable disinfectant
Tongs and dust pan
Spill Procedures
Place absorbent material over spill
Pour disinfectant around outer edges of absorbent ending in the middle
Wait for inactivation of bio-hazardous material
Properly dispose of all materials in biohazard box

25. Biohazardous Waste Includes...
Blood and Blood Products
Cultures/Stocks of Infectious Agents and Associated Biologicals
Animal Carcasses and Bedding
Sharps
Glass / Volumetric Pipettes
Primary and Established Human and Animal Cell Lines

26. BL1 and BL2 Waste Disposal Procedures
Liquid:
chemically treated prior to drain disposal
10% (total volume) sodium hypochlorite solution (Bleach; Hg free) for minutes
Solid:
BL1 and BL2 non recombinant waste
Dispose of directly into red plastic waste containers or cardboard boxes lined with red biohazard bags
Recombinant DNA
Any viable organism containing recombinant DNA must be rendered noninfectious prior to disposal
Steam sterilization or chemical inactivation

27. BL2S Waste Disposal Liquid Solid
Inactivate all liquid waste with Vesphene then autoclave
Solid
Autoclave all solid waste
Avoid using Sharps and Glass if possible

28. Sharps Disposal
All sharps should be disposed of directly into an approved sharps container immediately after use
When the sharps container is 2/3 full:
Place in biohazard waste container
Arrange for pick up from Building Services (x2656)

29. COMS Committee on Microbiological Safety
The MGH Institutional Biosafety Committee (IBC)
What research needs COMS approval
Infectious microorganisms, in-vitro or in-vivo
Recombinant DNA materials
Human studies involving rDNA (Human Gene Transfer studies)
Animal to human transplants (xenotransplantation)
Primate tissues
Dual use agents
All COMS approved research is subject to an annual biosafety inspection
The Forms:

30. OSHA Bloodborne Pathogens Training

31. Bloodborne Pathogens
What are the three bloodborne pathogens you need to be concerned about and what is the risk of transmission each?

32. BLOODBORNE PATHOGENS HEPATITIS B (HBV) 30% HEPATITIS C (HCV) 3 %
HIV %

33. Bloodborne Pathogens Hepatitis B Hepatitis C HIV
the most highly transmissible
but greatly reduced by vaccine use!!
Hepatitis C
No vaccine, but early identification and treatment offers some hope
HIV
Early treatment greatly reduces the risk

34. Exposure Prevention strategies
Vaccination
Work Practices/Standard Precautions
PPE/Engineering Controls
Exposure response
Education/polices

35. Hepatitis B vaccine
Available from Occupational Health Service (6-2217)
Must be offered to all employees with exposure risk
A 3 shot series- must complete
Antibody status checked after series completed
No need for routine “boosters”

36. Work Practice Controls
Standard Precautions
- Treat all blood and body fluids as infectious!
- Gloves for contact with blood, body fluids,
moist mucous membranes, non-intact skin,
secretions and contaminated items
- Other Personal Protective Equipment (PPE) as
needed
to suit the task you are performing
- Remove PPE when task completed and moving to a
clean area
- Disinfect hands after contact with blood/body
fluids/tissue, even if gloves are used
- Disinfect equipment/surfaces per policy

37. PPE/Engineering Controls
Gowns, gloves, face shields, splashguards etc.
Labeling
Biohazardous Waste Disposal
Safety Devices
Sharps Disposal
Sharps Containers

38. Post Exposure Response
How can you be exposed?
-Percutaneous
-Mucous membrane
-Broken Skin
- Animal Bites
Immediate Response
-First Aid- wash area
Reporting- Occ. Health (x2438) or call operator (x2000), available 24/7
-All animal bites must be reported to Occ. Health
Follow-up required

39. Policies and Procedures
Bloodborne Pathogens Exposure Control Plan- located in the Infection Control Manual
Departmental Policies
Annual Education