APPENDIX B

Recommended Emergency Procedures in Biohazard Laboratories

Note: Because of the diversity of projects and facilities, these procedures may not be applicable to all laboratories. The principal investigator is responsible for preparing written emergency procedures for his/her laboratory.

Introduction

Dropping culture flasks and Petri dishes in the open laboratory or containment failure can result in a significant release of aerosols and overt exposure of personnel to biohazardous agents. Laboratory personnel should be thoroughly familiar with emergency and decontamination procedures to minimize the extent and possible effects of exposure to hazardous agents.

Responsibility

The PI or lab manager is responsible to insure the use of proper cleanup procedures and containment.

Procedure

Containment Equipment Failure

  1. Stop work with potentially hazardous material should containment equipment or facility safeguards, such as a biological safety cabinet or building ventilation equipment, fail during an experiment,
  2. Notify the Principal Investigator and the University Biological Safety Officer at 515-6858

    3. Biohazardous Spills in an Open Laboratory



    Immediate Action

    1. Accidents and spills
      1. Clear the area at once
      2. Shut down air conditioning or ventilation to area if possible
      3. Notify the supervisor and laboratory director
      4. Assess the type of spill and degree of hazard involved
      5. Determine most effective and least hazardous approach to clean up and decontamination
      6. Determine the necessity for prophylactic treatment or other medical attention for persons exposed to the potentially hazardous materials

      Note: Laboratories handling highly virulent infectious agents must have specific plans for handling accident. Procedures for such circumstances are beyond the scope of this manual. The reader is referred to other sources such as DHEW publication CDC 74-8118, "Laboratory Safety at the Center for Disease Control", Center for Disease Control, Atlanta, Georgia.

    2. "Dry" spills (overturned or broken culture plate) with no significant aerosol formation
      1. Evacuation of room probably is not justified
      2. Flood area with disinfectant solution
      3. Soak up the disinfectant and contaminated material with an absorbent material ( sand, kitty litter, commercial type paper towels) and dispose of in a plastic bag or sealed container. Gloves should be worn for clean up.
      4. Autoclave the contaminated material.
    3. Liquid spills on bench or floor
      1. If significant aerosols were formed the area should be evacuated and not reentered for at least an hour.
      2. Cover the spill with an absorbent (sand, kitty litter, paper towels). Gloves should be worn during cleanup.
      3. Dispose of the absorbent and contaminated materials in plastic bags or sealed container and autoclave.
    4. Centrifuge accidents
      1. Shut off instrument, evacuate area at once, and shut down ventilation to area.
      2. Do not enter area for at least one hour or until aerosols have settled.
      3. The person entering the area to clean up will wear protective clothing, gloves, and mask.
      4. Soak up liquids in absorbent material and handle as above. If not a liquid, clean the instrument and clean the room thoroughly before resuming work.
    5. Spills in incubators, autoclaves, or other closed areas.
      1. Soak up liquids with an absorbent and dispose of as outlined in B-3 and B-4.
      2. The unit may need decontaminating by means of a sterilizing gas such as formaldehyde or glutaraldehyde and left overnight if routine cleanup is not possible.
      3. The unit should be thoroughly washed (if possible) after decontamination


    Decontamination Procedures

    1. Wait 30-60 minutes to allow dissipation of aerosols created by the spill.
    2. A "Spill Kit" should be available and should include leakproof containers and autoclavable bags, forceps, paper towels, sponges, disinfectant, respirators, and rubber gloves. A UV lamp may be helpful in emergency situations.
    3. Wear protective clothing when entering the laboratory to clean the spill area such as rubber gloves, autoclavable footwear, an outer garment, and a respirator. Do not wear a gown that may trail the floor when bending down for spills on the floor
    4. Pour a germicidal solution (10% Wescodyne in 50% ethanol or 5% Hypochlo- rite are recommended) around the spill and allow to flow into the spill. Paper towels soaked with germicide may be used to cover the area. To minimize re- aerosolization, avoid pouring the germicidal solution directly onto the spill.
    5. Let stand 20 minutes to allow adequate disinfectant contact time.
    6. Transfer contaminated materials (paper towels, glass, liquid, gloves, etc.) into a deep autoclave pan using an autoclavable dust pan and squeegee,. Cover the pan with aluminum foil or other suitable cover and autoclave according to standard directions.
    7. The dust pan and squeegee should be placed in an autoclavable bag and autoclaved according to standard directions.
    8. Remove protective clothing and sterilize by autoclaving.


    Biohazardous Spill in a Class II Biological Safety Cabinet

    1. Notify the Principal Investigator and the Biological Safety Officer if a splash occurrs outside the cabinet resulting in personnel exposure to hazardous material.
    2. Remove contaminated clothing and containerize it for autoclaving.
    3. Thoroughly wash hands and face if exposure has occurred.
    4. Initiate chemical decontamination procedure at once while the cabinet contin- ues to operate to prevent escape of contaminants from the cabinet.
    5. The operator will wear gloves and laboratory coat during this procedure.
    6. Spray or wipe walls, work surface, and equipment with an appropriate disinfectant detergent.
    8. Dump excess disinfectant from tray and drain pans into cabinet base.
    9. Discard gloves, cloth or sponge in an autoclave pan and autoclave.
    10. Drain disinfectant from cabinet base into an appropriate container and autoclave according to standard procedures.
    11. This procedure does not decontaminate the interior parts of the cabinet such as the filters, blowers, and air ducts. If the entire cabinet is to be decontami- nated with formaldehyde gas, contact the Biological Safety Officer at 515- 6858


    Radioactive-Biohazardous Spill

    1. The immediate actions are the same as for a biohazardous spill.
    2. The decontamination procedure can proceed in a manner similar to biohazard spill depending on an evaluation of the risk assessment of relative biological and radiological hazard..
    3. The Radiation Protection Officer will determine whether the contami- nated material should be autoclaved or chemically disinfected.
    4. The inactivated material should then be handled as radioactive waste.
    5. The final radioactive survey should be made of the spill area, clean-up tools, and contaminated clothing with a Geiger counter, or a smear should be taken and counted in a liquid scintillation counter. Twice (2X) background count indicates the need for further decontamination.


    Reporting Laboratory Accidents

    Report, in writing, to the Biological Safety Officer all laboratory accidents that result in exposure of laboratory personnel. Such exposures include inoculation through cutaneous penetration, ingestion, probable inhalation following gross aerosolization, or any incident causing serious exposure to personnel or danger of environmental contamination. The following information must be in the report

    1. Exposed personnel: Name, age, sex, occupation, department, and principal investigator
    2. Date, time, and place of exposure.
    3. Type of accident (accidental injection with a needle, contaminated aerosol, animal bite, etc.)
    4. Description of incident (specify hazardous agents)
    5. Medical attention provided.
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