2.0 Biosafety Level 2
This class of laboratory is suitable for work involving agents of moderate potential
hazard to personnel and the public. In addition to the characteristics of a Biosafety
Level 1 facility, (a) laboratory personnel have specific training in handling pathogenic
agents and are directed by competent scientists, (b) access to the laboratory is limited
whenever work is being conducted, and (c) when some procedures are conducted in which
infectious aerosols are created, they will be performed in biological safety cabinets
or other physical containment equipment.
Standard Microbiological Practices
- Restrict access to the laboratory, at the discretion of the laboratory director, when
experiments are in process.
- Decontaminated work surfaces once each day and after any spill of viable material.
- Decontaminate all contaminated liquid or solid wastes before disposal.
- Always use mechanical pipetting devices; mouth pipetting is prohibited.
- Eating, drinking, smoking and applying cosmetics are not permitted in the work area.
Food can only be stored in cabinets or refrigerators specifically designated for that
purpose. These must be located outside the work area.
- Wash hands with a suitable disinfectant after handling viable materials and before
leaving the laboratory.
- Perform all procedures in a way that minimizes the creation of aerosols.
Special Practices
- Place all contaminated materials that will be decontaminated at a site away from the
laboratory in a durable, leakproof container, which is closed before being removed
from the laboratory.
- Limit access to the laboratory for persons who are at increased risk of acquiring
infection, or for whom infection might be unusually hazardous. The Laboratory Director
has the final responsibility for determining who may enter or work in the laboratory.
- Establish written policies and procedures whereby only persons who have been advised
of the hazards and meet any specific entry requirements, e.g., special training or
immunization, may enter the facility.
- Post special provisions for entry, e.g., vaccination, a hazard warning sign incorporating
the universal biohazard symbol on the door to the laboratory work area when the infectious
agents is used in the laboratory . The warning sign identifies the infectious agent,
lists the name of the Laboratory Director, laboratory supervisor and Department Head,
and identifies the special requirements for entering the laboratory.
- Ensure that an effective insect and rodent control program is in effect.
- Wear laboratory coats, gowns, smocks or uniforms while in the laboratory. Before leaving
the laboratory remove these garments and leave them in the laboratory. A clean coat
may be worn over the potentially contaminated garments, but the outer garment must
be considered contaminated upon removal.
- Do not permit animals not involved in the research in the laboratory.
- Use special care to avoid skin contamination with infectious materials. Wear gloves
when handling infected animals and when skin contact with infectious materials is
unavoidable.
- Contaminate all wastes from laboratories and animal rooms before disposal.
- Use hypodermic needles and syringes only for parenteral injection and aspiration of
fluids from laboratory animals and diaphragm bottles. Use only needle-locking syringes
or disposable syringe-needle units for the injection or aspiration of infectious fluids.
Extreme caution should be used when handling needles and syringes to avoid autoinoculation
and the generation of aerosols during use and disposal. Do not bend, cut, or replace
needles in the sheath or guard or removed them the syringe following use. Place the
needle and syringe promptly in a puncture-proof container and decontaminated, preferably
by autoclaving, before discard or reuse.
- Report spills and accidents that result in overt exposures to infectious agents immediately
to the Laboratory Director. Medical evaluation, surveillance, and treatment will be
provided as needed at no cost to the employee. Maintain written records of the incident,
including a determination of the causes, any personnel exposed, and any actions that
were taken.
- Collect baseline serum samples for those individuals working with materials infectious
to humans (including possibly infectious animals, animal tissues that have not been
rendered safe, or derivative materials) and store. Additional serum samples will be
taken at five-year intervals or after an overt exposure. Also, at a minimum, a medical
history is required and it is recommended that the employees participate in a full
Health Assurance Program.
- Prepare a biosafety procedures. Advise personnel of special hazards that are required
to become familiar with experimental practices and procedures, including all safety
precautions.
Containment Equipment
- Use Class I or II biological safety cabinets or other appropriate personal protective
or physical containment devices in the following situations:
- When procedures with a high potential for creating infectious aerosols are conducted.
These include but are not limited to centrifuging, grinding, blending, vigorous shaking
or mixing, sonic disruption, pipetting, opening pressurized containers, inoculating
animals intranasally, and harvesting infected tissues from animals, eggs, or plants.
- When high concentrations or large volumes of infectious agents are used. Such materials
may be centrifuged in the open laboratory if sealed heads or centrifuge safety cups
are used and if they are opened only in a biological safety cabinet.
Laboratory Facilities
- Ensure that the laboratory is designed so that it is easily cleaned. Seamless or poured
floor coverings are recommended. Epoxy paint is recommended for the walls. Junctures
with the wall, floor, ceiling, and equipment should be rounded, with no corner seams.
Laboratory furniture should incorporate as few sharp seams and cracks as possible.
Penetrations in the floor should be sealed or capable of being sealed to aid decontamination.
- Bench tops must be impervious to water and resistant to acids, alkalis, organic solvents,
and moderate heat.
- Spaces between benches, cabinets, and equipment must be accessible for cleaning.
- Each laboratory contains a sink for hand washing.
- Equip windows that can be opened with fly screens.
- An autoclave should be available for decontaminating infectious wastes.