Biosafety Level (BSL) Practices Chart
Last Updated: March 2, 2024 7:24:12 PM PST
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See UC San Diego's laboratory containment requirements and practices for biosafety levels 1 through 3 in the chart below.
Biosafety Levels (BSL)
Research and teaching activities involving infectious agents requires prior approval by the Institutional Biosafety Committee (IBC) via the Biohazard Use Authorization (BUA) review process.
Each laboratory space where biohazardous materials are used is assigned one of 3 internationally recognized biosafety levels or BSL. The biosafety level is commensurate with the:
- Degree of risk posed by the biohazardous materials
- Activities carried out with those materials
The IBC uses the biosafety levels recommended by the CDC and NIH as the usual standards of containment to be set for work with a given biohazardous material. Containment requirements are subject to modification by the IBC at its discretion, depending on the circumstances presented by a specific project.
*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.
Follow requirements and practices for your assigned BSL:
| Biosafety levels (BSL) | BSL–1 | BSL–2 |
BSL–2+ *See agents that require enhanced precautions (BSL-2+) |
BSL–3 | References |
| A. Hazard levels | |||||
| 1. 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. | High: Agents that cause disease of moderate to high hazard that have serious or potentially lethal consequences | Biosafety in Microbiological and Biomedical Laboratories, Centers for Disease Control and Prevention (CDC) |
| 2. Examples | Escherichia coli (laboratory strain) |
Streptococcus pyogenes |
Listeria monocytogenes |
Mycobacterium tuberculosis | |
| B. Standard microbiological practices | |||||
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Biosafety levels (BSL)
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BSL–1
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BSL–2 |
BSL–2+ *See agents that require enhanced precautions (BSL-2+) |
BSL–3
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References
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| 1. Access to the laboratory | Access does not have to be restricted – however, doors cannot be propped open (in violation of fire code). | Doors to the laboratory are closed when BSL-2 work is being conducted to prevent public access. | Doors to the laboratory are closed when BSL-2 work is being conducted to prevent public access. | Doors to the laboratory are closed and locked to prevent untrained personnel access. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 2. Biohazard signage | Biohazard sign must be posted. | A biohazard symbol must be posted on the laboratory room sign. Large equipment where biohazardous materials are used and/or stored, such as biosafety cabinets, incubators, refrigerators and freezers, must be visibly labeled with the universal biohazard symbol. | Biosafety in Microbiological and Biomedical Laboratories, CDC | ||
| 3. Biohazard solid waste decontamination | Biomedical waste vendor. | 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. | Steam sterilize in laboratory – EH&S may grant exceptions in extenuating circumstances. | Biohazardous Waste Disposal Guidelines (PDF) |
| 4. Biohazardous liquid culture decontamination | 10% bleach/water made fresh daily with bleach having an EPA registration number (e.g., Chlorox) for 30 minutes. | 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. | Steam sterilize in laboratory – EH&S may grant exceptions in extenuating circumstances. | How to Disinfect Tissue Culture Media in Vacuum Flasks |
| 5. 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. | Not permitted at any time. | Laboratory Clean Areas |
| 6. Contaminated sharps (e.g., needles, blades, glass) | Safe handling practices must be developed and implemented. Substitute plasticware for glassware whenever possible. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 7. Decontamination of work surfaces | Daily, after finishing work and following spills. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 8. Pipetting | Mechanical device – no mouth pipetting. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 9. 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. | Biohazardous Waste Disposal Guidelines (PDF) | |||
| 10. Handwashing | Required after working with potentially hazardous materials and before leaving the laboratory. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 11. Training | The laboratory supervisor must ensure that laboratory personnel |
Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 12. 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. |
Biosafety in Microbiological and Biomedical Laboratories, CDC Vaccination for Researchers and Research Support Staff & Medical Consults |
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| 13. Equipment decontamination | Equipment must be cleaned of residues and green tagged by EH&S before repair, maintenance, or removal from laboratory. | Equipment must be decontaminated and green tagged by EH&S before repair, maintenance, or removal from laboratory. | How to Get a Decontamination Clearance for Equipment or Facilities | ||
| 14. Animals and plants not associated with the work | Allowed if approved by laboratory director and university policy. | Not allowed in the laboratory. | Biosafety in Microbiological and Biomedical Laboratories, CDC | ||
| 15. Institutional Biosafety Committee approval required for use of glassware | Not applicable. | Recommended | Required for HBV, HCV, and HIV | Required. | Institutional Biosafety Committee |
| C. Safety equipment | |||||
| Biosafety levels (BSL) | BSL–1 | BSL–2 |
BSL–2+ *See agents that require enhanced precautions (BSL-2+) |
BSL–3 | References |
| 1. Class II Biological safety cabinet (with annual certification) | Not required. | Required for all aerosol generating processes** | Required for all work. | Required for all work. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 2. 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. | Required for all work. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 3. Laboratory coats | Required. | Required. | Required. | Required (solid front disposable gown). | Laboratory Hazard Assessment Tool |
| 4. Gloves (alternatives to latex gloves should be available) | Required. | Required. | Required. | Required. | Laboratory Hazard Assessment Tool |
| 5. 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. | Required. This includes work in the biosafety cabinet. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 6. Sleeve protectors | Not required. | Recommended. | Recommended. | Institutional Biosafety Committee | |
| 7. HEPA-filtered vacuum lines | Required. | Institutional Biosafety Committee | |||
| D. Laboratory facilities | |||||
| Biosafety levels (BSL) | BSL–1 | BSL–2 |
BSL–2+ *See agents that require enhanced precautions (BSL-2+) |
BSL–3 | References |
| 1. Ventilation | Negative pressure is required if adjacent area is a lower biosafety level or non-laboratory space. Single-pass air is required. Reference: Institutional Biosafety Committee | ANSI/AIHA Z9.5-2002, Laboratory Ventilation, August 29, 2002, Chapter 5, Laboratory Ventilation System Design. UC Laboratory Safety Design Guide, July 2004, Chapter 3, Laboratory Ventilation and Fume Hoods. |
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| 2. Handwashing facilities | Required. | Required (foot/elbow/electronic operation) | Biosafety in Microbiological and Biomedical Laboratories, CDC | ||
| 3. Autoclave | Not required. | Required in laboratory. | Biosafety in Microbiological and Biomedical Laboratories, CDC | ||
| 4. Eye wash station | Recommended. However, use of hazardous chemicals may change this to a requirement. | Required. | Required | Required in laboratory. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 5. Doors | Required. | Required. Doors should be self-closing and have locks. | Required. Doors should be self-closing and have locks. | Required. A series of 2 self-closing doors is the basic requirement for entry. The space between the 2 doors is called the anteroom. Palm scanners are used to restrict access. | Biosafety in Microbiological and Biomedical Laboratories, CDC |
| 6. Chairs | Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |||
| 7. Cleaning and decontamination | Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate. | Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate. | Laboratory design should allow the facility to be easily cleaned and decontaminated. Carpets and rugs are not appropriate. Seams, floors, walls, and ceiling surfaces should be sealed. Spaces around doors and ventilation openings should be capable of being sealed to allow for space fumigation. | Biosafety in Microbiological and Biomedical Laboratories, CDC | |
* Agents that require enhanced precautions (BSL-2+)
The Institutional Biosafety Committee (IBC) has approved specific Exposure Control Plans (ECP) that require additional precautions and safety equipment for certain agents (see the list below). Handling macaque monkeys or their blood, tissue, or body fluids requires using the macaque ECP.
| Agents that require enhanced precautions (BSL-2+) | |
|---|---|
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Any exceptions should be approved by UC San Diego Biosafety or the IBC.
** 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
Contact EH&S Biosafety.
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