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Safe Laboratory Practices
Laboratory Biosafety Level Criteria
Table1: Summary of  Recommended Biosafety  Levels for Infectious Agents
Table1a: Summary of Vertebrate Animal Biosafety Levels (ABSL)
Biological Emergencies
Biosafety Cabinets (BSC's)
Types of BSC's
Comparison of BSC's Characteristics
Selection of a BSC Through Risk Assessment
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Animal Biosafety Level 2 (ABSL2)

Animal Biosafety Level 2 involves practices for work with those agents associated with human disease. It addresses hazards from ingestion as well as from percutaneous and mucous membrane exposure. ABSL-2 builds upon the practices, procedures, containment equipment, and facility requirements of ABSL-1.

A. Standard Practices
  1. Aside from the standard policies, procedures, and protocols for emergency situations established by the facility director, appropriate special policies and procedures should be developed as needed and approved by the Institutional Animal Care and Use Committee (IACUC) and the Institutional Biosafety Committee (IBC).

  2. Access to the animal room is limited to the fewest number of individuals possible. Personnel who must enter the room for program or service purposes when work is in progress are advised of the potential hazard.

  3. An appropriate medical surveillance program is in place.   All personnel receive appropriate immunizations or tests for the agents handled or potentially present (e.g., hepatitis B vaccine, TB skin testing).  When appropriate, a serum surveillance system should be implemented.

  4. A biosafety manual is prepared or adopted.  Personnel are advised of special hazards, and are required to read and follow instructions on practices and procedures.

  5. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use should only be done in designated areas and are not permitted in animal or procedure rooms.

  6. All procedures are carefully performed to minimize the creation of aerosols or splatters.

  7. Equipment and work surfaces in the room are routinely decontaminated with an effective disinfectant after work with the infectious agent, and especially after overt spills, splashes, or other contamination by infectious materials.

  8. All infectious samples are collected, labeled, transported, and processed in a manner that contains and prevents transmission of the agent(s).  All wastes from the animal room (including animal tissues, carcasses, contaminated bedding, unused feed, sharps, and other refuse) are transported from the animal room in leak-proof, covered containers for appropriate disposal in compliance with applicable institutional or local requirements.  The outer surface of the containers is disinfected prior to moving the material.  Autoclaving of the contents prior to incineration is recommended.

  9. Policies for the safe handling of sharps are instituted:
    1. Needles and syringes or other sharp instruments are restricted for use in the animal facility only when there is no alternative, such as for parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles.

    2. Syringes that re-sheathe the needle, needle-less systems, and other safe devices should be used when appropriate.

    3. Plastic ware should be substituted for glassware whenever possible.

  10. Personnel wash their hands after handling cultures and animals, after removing gloves, and before leaving the animal facility.

  11. A biohazard sign must be posted on the entrance to the animal room whenever infectious agents are present. The hazard warning sign identifies the infectious agent(s) in use, lists the name and telephone number of the responsible person(s), and indicates the special requirements (e.g., the need for immunizations and respirators) for entering the animal room.

  12. An insect and rodent control program is in effect (see BMBL 4th Edition: Appendix G).

B. Special Practices
  1. Animal care laboratory and support personnel receive appropriate training on the potential hazards associated with the work involved, the necessary precautions to prevent exposures, and the exposure evaluation procedures.  Personnel receive annual updates, or additional training as necessary for procedural or policy changes.   Records of all training provided are maintained.  In general, persons who may be at increased risk of acquiring infection, or for whom infection might be unusually hazardous, are not allowed in the animal facility unless special procedures can eliminate the extra risk.

  2. Only animals used for the experiment(s) are allowed in the room.

  3. All equipment must be appropriately decontaminated prior to removal from the room.

  4. Spills and accidents which result in overt exposures to infectious materials must be immediately reported to the facility director.  Medical evaluation, surveillance, and treatment are provided as appropriate and written records are maintained.
C. Safety Equipment (Primary Barriers)
  1. Gowns, uniforms, or laboratory coats are worn while in the animal room. The laboratory coat is removed and left in the animal room.  Gowns, uniforms, and laboratory coats are removed before leaving the animal facility.  Gloves are worn when handling infected animals and when skin contact with infectious materials is unavoidable.

  2. Personal protective equipment is used based on risk assessment determinations.  Appropriate face/eye and respiratory protection is worn by all personnel entering animal rooms that house nonhuman primates.

  3. Biological safety cabinets, other physical containment devices, and/or personal protective equipment (e.g., respirators, face shields) are used whenever conducting procedures with a high potential for creating aerosols.  These include necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, or intranasal inoculation of animals.

  4. When needed, animals are housed in primary biosafety containment equipment appropriate for the animal species.  Filter top cages are always handled in properly designed and operating animal biocontainment cabinets recommended for rodents.
D. Facilities (Secondary Barriers)

  1. The animal facility is separated from areas that are open to unrestricted personnel traffic within the building.

  2. Access to the facility is limited by secure locked doors.  External doors are self-closing and self-locking.   Doors to animal rooms open inward, are self-closing, and are kept closed when experimental animals are present.   Cubicle room inner doors may open outward or be horizontal or vertical sliding.

  3. The animal facility is designed, constructed, and maintained to facilitate cleaning and housekeeping.  The interior surfaces (walls, floors, and ceilings) are water resistant.

  4. Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, are arranged to minimize horizontal surface areas.

  5. Windows are not recommended. Any windows must be resistant to breakage and should be sealed.

  6. If floor drains are provided, the traps are always filled with an appropriate disinfectant.

  7. Exhaust air is discharged to the outside without being recirculated to other rooms.  Ventilation should be provided in accordance with criteria from Guide for Care and Use of Laboratory Animals, latest edition.  The direction of airflow in the animal facility is inward; animal rooms should maintain negative pressure compared to adjoining hallways.

  8. Cages are washed manually or in an appropriate cage washer.   The mechanical cage washer should have a final rinse temperature of at least 180F.

  9. An autoclave is available in the animal facility to decontaminate infectious waste.

  10. A hand washing sink is in the animal room where infected animals are housed, as well as elsewhere in the facility.

  11. Illumination is adequate for all activities, avoiding reflections and glare that could impede vision.

ABSL1 ABSL3 ABSL2

Reproduced from "Biosafety in Microbiological and Biomedical Laboratories, BMBL 4th Edition" with permission from the Center for Disease Control (CDC).
     
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