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Biosafety in Research
Laboratories
1
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
What is Biohazardous Material?
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Bacteria
Viruses
Parasites
Fungi
Human and Non-Human Primate Material
Recombinant DNA
Animals
Biological Toxins
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
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
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Lab Coat / gown, gloves, goggles, facial protection
Close toed shoes
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)
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
Safety Equipment
(Primary Barriers)
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Biosafety cabinets (BSCs)
Safety centrifuge cups and rotors
Pipetting Devices
Vacuum line HEPA Filters
Personal protective clothing
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Gloves (remember the one glove rule)
Gowns/Lab coats (should remain inside the lab)
Eye and face protection
Close toed shoes
Biological Safety Cabinets
Must be Certified Annually
•
Air Systems Technologies
(508) 427 5558
BSC Provide:
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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
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
Centrifuges
 Safety Cups and
Rotors
Load and unload
safety cups inside the
BSC
Vacuum Line HEPA Filter Protection
 Placed in between the
vacuum system and
the aspiration flasks
Biosafety Levels
 BSL-1:
 BSL-2:
 BSL-3:
 BSL-4:
agents not known to cause disease (in
healthy adults)
agents associated with disease
indigenous/exotic agents
associated with human disease and
with potential for aerosol
transmission
dangerous/exotic agents of life
threatening nature
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)
Biosafety Level 1
Immunocompromised workers are at risk
Standard Operating Procedures
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Minimize splashes and aerosols
PPE
Decontaminate work surfaces daily
Decontaminate wastes
Maintain insect & rodent control program
Examples:
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Adeno associated virus
Murine leukemia virus
Non pathogenic E. coli
Animal tissue and cells (other than NHP)
Biosafety Level 1
Laboratory Facilities (Secondary
Barriers)
Biosafety Level 2
• Infectious dose will vary based on:
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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
Biosafety Level 2
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Biosafety Level 2 Signs
• Policies and
procedures for entry
• Biohazard warning
signs
Specific hazard training with
annual updates
Biosafety Level 2
Laboratory Facilities (Secondary Barriers)
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Hand washing sink
located near laboratory
exit
Autoclave available
Eyewash/Shower
station available
Biosafety Cabinets
Biosafety Level 2
Laboratory Facilities (Secondary Barriers)
Special Practices
Use leak-proof transport containers
Biological Spill Kits
• Location
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Inside Tissue Culture Rooms
• Contents
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Gloves, Goggles/shield, N95 Respirator, Lab coat/gown
Absorbent material
Bleach or suitable disinfectant
Tongs and dust pan
• Spill Procedures
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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
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
BL1 and BL2 Waste Disposal
Procedures
 Liquid:
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chemically treated prior to drain disposal
10% (total volume) sodium hypochlorite solution (Bleach; Hg
free) for 20-30 minutes
 Solid:
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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
BL2S Waste Disposal
• Liquid
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Inactivate all liquid waste with Vesphene then
autoclave
• Solid
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Autoclave all solid waste
Avoid using Sharps and Glass if possible
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)
COMS
• Committee on Microbiological Safety
•
The MGH Institutional Biosafety Committee (IBC)
• What research needs COMS approval
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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:
http://www.hms.harvard.edu/orsp/coms/
OSHA Bloodborne Pathogens
Training
Bloodborne Pathogens
What are the three
bloodborne
pathogens you
need to be
concerned about
and what is the
risk of
transmission each?
BLOODBORNE PATHOGENS
 HEPATITIS B (HBV)
30%
 HEPATITIS C (HCV)
3%
 HIV
0.3%
Bloodborne Pathogens
 Hepatitis B
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
Exposure Prevention strategies
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Vaccination
Work Practices/Standard Precautions
PPE/Engineering Controls
Exposure response
Education/polices
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”
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
PPE/Engineering Controls
 Gowns, gloves, face shields, splashguards
etc.
 Labeling
 Biohazardous Waste Disposal
 Safety Devices
 Sharps Disposal
Sharps Containers
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
Policies and Procedures
 Bloodborne Pathogens Exposure Control
Plan- located in the Infection Control
Manual
http://library.partners.org/MGH1/webserver/custom/tr
ov
 Departmental Policies
 Annual Education