Biosafety Levels (BSL), Risk, and Containment

IBC Standard Operating Procedure

The IBC uses the biosafety levels recommended by the CDC and NIH as the usual standards of containment*.  The biosafety level is based on the

• Degree of risk posed by the biohazardous materials
• Activities carried out with those materials

*Containment is the term used to describe methods, practices, procedures, facilities, and equipment used to safely manage biohazardous materials in the laboratory.  The purpose of containment is to reduce or eliminate exposure to people or the environment to potentially hazardous agents.

	BSL-1	BSL-2	BSL-2+ (BL2 enhanced)
HAZARD LEVELS
Degree of hazard	Low risk - well characterized agents not known to cause disease in healthy adult humans.	Moderate - agents that cause human disease of moderate hazard.	High - agents involved in laboratory-acquired infections or where disease can have serious or potentially lethal consequences.
Examples	Escherichia coli (laboratory strain); all exempt r/sNA work.	Streptococcus pyogenes; Adenovirus all Types; Human cell lines, eg. HEK 293; Herpes Simplex Virus; Rabies Virus; Pseudorabies Virus	See Agents that Require Enhanced Precautions**
STANDARD MICROBIOLOGICAL PRACTICES
Access to the laboratory	Access does not have to be restricted. Doors cannot be propped open (fire code violation).	Doors to the laboratory are closed when BSL-2 work is being conducted to prevent public access.
Signage	Biohazard sign must be posted.
Training	The laboratory supervisor must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures.
Medical surveillance	Recommended where personal health status may result in increased susceptibility to infection or inability to receive vaccinations or prophylactic interventions.	Required. Laboratory personnel must be provided with medical surveillance and offered appropriate immunizations.
Eating, drinking, application of cosmetics or contact lenses	Permitted only in designated clean areas.	Permitted only in designated clean areas. Not permitted if Aerosol Transmissible Disease Pathogens are used.	Not permitted at any time.
Handwashing	Required after working with potentially hazardous materials and before leaving the laboratory.
Pipetting	Mechanical device – no mouth pipetting.
Contaminated sharps (needles, blades, glass)	Safe handling practices must be developed and implemented. Substitute plasticware for glassware whenever possible.
IBC approval required for use of glassware	Not applicable.	Recommended.	Required for HBV, HCV, and HIV.
Storage of biohazardous waste material	Double red bags held in rigid, leakproof containers with biohazard labels on the top and side. Biohazardous waste must be under direct control of the responsible laboratory until it is placed in an EH&S approved storage area.
Biohazard solid waste decontamination	Biomedical waste vendor or steam sterilize with EH&S approval.		Biomedical waste vendor or steam sterilize with EH&S approval. Pathological waste or infected animals must be incinerated.
Biohazardous liquid culture decontamination	10% bleach/water made fresh daily with bleach having an EPA registration number (e.g., Chlorox) for 30 minutes or steam sterilize with EH&S approval.		10% bleach/water made fresh daily with bleach having an EPA registration number (e.g., Chlorox) for 30 minutes or steam sterilize with EH&S approval. Prion liquid waste must be added to 1 Normal NaOH and collected as hazardous waste.
Equipment decontamination	Equipment must be cleaned of residues and tagged by EH&S before repair, maintenance, or removal from laboratory.	Equipment must be decontaminated and tagged by EH&S before repair, maintenance, or removal from laboratory.
Work surface Decontamination	Daily, after finishing work and following spills, using 10% bleach solution, 70% alcohol or approved cleaning product.
Animals and plants not associated with the work	Allowed if approved by laboratory director and university policy.	Not allowed in the laboratory.
SAFETY EQUIPMENT
Class II Biological safety cabinet (annual certification)	Not required.	Required for all Aerosol Generating Processes***.	Required for all work.
Sealed rotors or safety cups for centrifuging	Not required.	Required for high concentrations or large volumes of infectious agents. Exception: centrifuges without secondary containment can be operated inside a certified biosafety cabinet.	Required for all work.
Laboratory coats	Required.	Required.	Required.
Gloves	Required.	Required.	Required.
Eye protection (safety glasses, goggles)	Required. This includes work in the biosafety cabinet.	Required. This includes work in the biosafety cabinet.	Required. This includes work in the biosafety cabinet.
Sleeve protectors	Not required.		Recommended.
HEPA-filtered vacuum lines	Required.
LABORATORY FACILITIES
Ventilation	Not required.	Use laminar
Autoclave	Not required.
Doors	Required.	Doors should be self-closing and have locks.
Handwashing facilities	Required.
Eye wash station	Recommended. However, use of hazardous chemicals may change this to a requirement.	Required.	Required
Chairs	Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant.
Cleaning and decontamination	Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate.

 ** Agents that require enhanced precautions (BSL-2+)

• Escherichia coli (shiga toxin producing strains)
• Hepatitis B Virus (HBV)
• Hepatitis C Virus (HCV)
• Human Immunodeficiency Virus (HIV)
• Influenza virus
• Infectious Prions 
• Listeria monocytogenes
• Lymphocytic Choriomeningitis Virus (LCMV)
• Macaque tissue
• Oncogenes used in viral vectors
• Rabies Virus or vectors
• Salmonella Typhi
• Shigella dysenteriae (Type 1)
• Simian Immunodeficiency Virus (SIV)
• Toxoplasma gondii
• Vaccinia virus or vector
• Zika virus

 *** Aerosol generating processes

• Centrifuging
• Grinding
• Blending
• Vigorous shaking or mixing
• Sonic disruption
• Opening containers with high internal pressures
• Inoculating animals intranasally
• Harvesting tissues from animals or embryonated eggs

References

• Biosafety in Microbiological and Biomedical Laboratories (BMBL, 5th ed.)
• Risk Group Database - American Biological Safety Association(ABSA)
• Pathogen Safety Data Sheets and Risk Assessment -Public Health Agency of Canada

 IBC Approved:  October 18, 2017