Biosafety Levels 1-4

• Biosafety Levels 1-4 include standard and special microbiological practices
• Biosafety Levels 1-4 include safety equipment and laboratory facilities
• These elements apply to activities involving infectious microorganisms, toxins, and laboratory animals
• The four levels of biosafety are organized in ascending order of protection for personnel, the environment, and the community
• Special practices address unique risks associated with handling agents requiring increasing containment levels
• Appropriate safety equipment and laboratory facilities enhance worker and environmental protection.
• Features of each Biosafety Level (BSL) are summarized in Table 1
• Adjustments to containment levels are risk-based, as detailed in Section II
• Worker safety and health concerns are coordinated with the Institutional Biosafety Committee (IBC)
• All hazards are addressed as part of the protocol review process
• Occupational health information is in Section VII.

Biosafety Level 1 (BSL-1)

• BSL-1 is suitable for work with well-characterized agents not known to consistently cause disease in immunocompetent adult humans
• BSL-1 presents a minimal potential hazard to laboratory personnel and the environment
• BSL-1 laboratories are not necessarily separated from general building traffic
• Work is typically conducted on open benchtops using standard microbiological practices
• Special containment equipment or facility design is not generally required
• Laboratory personnel receive specific training in procedures and are supervised by a scientist with training in microbiology or a related science
• Standard practices, safety equipment, and facility specifications are recommended for BSL-1.

Standard Microbiological Practices for BSL-1

• The laboratory supervisor enforces institutional policies that control safety and access.
• The laboratory supervisor ensures personnel receive training on duties, hazards, manipulations, precautions, and hazard evaluations
• Appropriate records are maintained.
• Personnel receive annual updates and training when equipment, procedures, or policies change
• All entering the facility are advised of potential hazards, instructed on safeguards, and follow practices and procedures
• Institutional policy regarding visitor training, occupational health, and safety communication is considered.
• Personal health status may affect susceptibility to infection and ability to receive immunizations or prophylactic interventions
• Personnel, especially those of reproductive age or with predisposing conditions, receive information on immune competence and susceptibility
• Those with such conditions are encouraged to self-identify to the institution’s healthcare provider. See Section VII.
• A facility-specific safety manual is prepared/adopted in consultation with the facility director and safety professionals
• The safety manual is available, accessible, periodically reviewed, and updated.
• The safety manual contains information describing biosafety and containment procedures
• The safety manual contains information for organisms/biological materials in use, agent-specific decontamination methods, and work performed
• The safety manual contains protocols for emergency situations, including exposures, emergencies, facility malfunctions, and other emergencies
• Training in emergency response procedures is provided according to institutional policies.
• A sign is posted at the entrance when infectious materials are present
• The sign should include the Biosafety Level, supervisor's name and number, PPE requirements, occupational health requirements, and entry/exit procedures
• Agent information is posted per institutional policy.
• Long hair is restrained to avoid contact with hands, specimens, containers, or equipment.
• Gloves are worn to protect hands from hazardous materials.
• Glove selection is risk-based.
• Gloves are not worn outside the lab.
• Change gloves when contaminated, if glove integrity is compromised, or when necessary.
• Do not wash/reuse disposable gloves; dispose of contaminated waste.
• Gloves and PPE are removed to minimize personal contamination and the transfer of infectious materials outside of contained/manipulated areas.
• Hands are washed after working with hazardous materials and before leaving the lab.
• Eating, drinking, smoking, handling contacts, applying cosmetics, and storing food are not permitted in laboratory areas.
• Food is stored outside.
• Mouth pipetting is prohibited; mechanical pipetting devices are used.
• There should be policies for safe handling of sharps: needles, scalpels, pipettes, and broken glassware that is consistent with state, federal, and local requirements
• Supervisors should adopt work practices to reduce sharps injuries
• Substitute plasticware for glassware when possible.
• Limit use of needles, syringes, and other sharps to situations where there is no alternative
• Use active/passive needle-based safety devices whenever possible.
• Uncapping of needles is performed in a way to reduce the potential for recoil causing an accidental needlestick.
• Needles are not bent, sheared, broken, recapped, removed from disposable syringes, or manipulated by hand before disposal.
• If necessary to remove a needle from a syringe (e.g., to prevent lysing blood cells) or recap, use hands-free device or comparable safety procedure (e.g., sharp container).
• Used, disposable needles and syringes are carefully placed in puncture-resistant sharps containers immediately after use close to the point of use.
• Non-disposable sharps are placed in hard-walled containers for transport to a processing area for decontamination, preferably autoclaving.
• Broken glassware is not handled directly; use a brush and dustpan, tongs, or forceps.
• Perform all procedures to minimize the creation of splashes and aerosols.
• Work surfaces should be decontaminated after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant
• Spills of infectious materials are contained, decontaminated, and cleaned by trained/equipped staff
• A spill procedure is developed and posted.
• Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method, consistent with requirements.
• When decontaminating outside of the immediate laboratory, materials are placed in a durable, leak-proof container and secured for transport
• Outer surface of containers holding infectious materials is disinfected and the container labeled with a universal biohazard label.
• Materials removed from the facility for decontamination are packed according to regulations.
• An effective integrated pest management program is implemented. See Appendix G.
• Animals and plants not associated with the work are not permitted in the laboratory.

Special Practices for BSL-1

• None required.

Safety Equipment for BSL-1

• Special containment devices or equipment, such as biosafety cabinets (BSCs), are not generally required.
• Protective laboratory coats, gowns, or uniforms are worn to prevent contamination of personal clothing.
• Protective eyewear is worn to prevent splashes and sprays of microorganisms/hazardous materials
• Eye/face protection are disposed of/decontaminated after use.
• When research animals are present, risk assessment considers appropriate eye, face, respiratory protection, and potential animal allergens.

Laboratory Facilities for BSL-1

• Laboratories have doors for access control.
• Laboratories have a sink for handwashing.
• An eyewash station is readily available.
• The laboratory is designed to be easily cleaned.
• Carpets and rugs are not appropriate.
• Spaces between benches, cabinets, and equipment should be accessible for cleaning.
• Laboratory furniture can support anticipated loads and uses.
• Benchtops are impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals.
• Chairs used in laboratory work are covered with a non-porous material that can be easily cleaned and disinfected.
• Laboratory windows that open to the exterior are fitted with screens.
• Illumination is adequate for all activities and avoids reflections and glare.

Overview of Biosafety Levels 1-4, including microbiological practices, safety equipment and lab facilities. The four levels of biosafety are organized in ascending order of protection for personnel, the environment, and the community. Special practices address unique risks.