21. Biological Spills

Since spills of biological materials will happen, it is important to be prepared prior to having to deal with the problem. Assess the spill! Is it a large spill or a small spill? A large spill is generally defined as a sufficient quantity that if spilled tends to seek its own level. In other words it runs to a low point. The main concept that would cause one to treat a large spill differently is containment: one would want to make sure the spill did not spread and contaminate other areas.

21.1 Biohazard Spill Cleanup Procedures

The following procedures are provided as a basic guideline to biohazardous spill cleanup, and will need to be modified for specific situations. As with any emergency situation: stay calm, call 2999 if necessary, and proceed with common sense. Call 064-710719 if further assistance is required, especially if the spill outgrows the resources in the laboratory.

Additional information regarding an emergency plan is included in Appendix F of CDC/NIH Biosafety in the Microbiological and Biomedical Laboratories.

21.2 Spill Kit

All laboratories working with biohazards should have a basic biological spill kit ready to use at all times. For most instances, the basic kit can be assembled with materials already available in the laboratory. Although it is preferable to have the contents of the spill kit in one location, as long as the materials are easily accessible to everyone in the lab, prior assembly might not be necessary.

The basic biological spill kit contains:

• Disinfectant (bleach, bleach 1:10 freshly prepared, or Iodine-based disinfectants)
• Absorbent material (e.g., paper towels)
• Waste containers (e.g., biohazard bags, sharp containers)
• Mechanical tools (e.g., household rubber gloves and forceps to deal with broken glass, dustpan and broom)
• Personal protective equipment (e.g., gloves, masks, goggles, disposable gowns)

21.3 Exposed Personnel:

• Eye splash: Use the eye wash for 15 minutes, holding the eye open.
• Hands or other exposed skin: Wash with antiseptic or soap.
• Report to your supervisor immediately for antiviral prophylaxis

21.4 Spills inside the Laboratory

• Notify others in the area, to prevent contamination of additional personnel and environment.
• Avoid inhaling airborne material, while quickly leaving the room. Notify others to leave.
• Close the door, and post a warning sign.
• Inform the supervisor.
• Allow aerosols to settle for at least 30 minutes before re-entering the laboratory.
• Have a complete biological spill kit ready to go before you start the cleanup.
• Put on protective clothing gown, aerosol type mask (for example 3M 9332), gloves, and shoe covers.
• Initiate cleanup with disinfectant as follows:
  1. Cover the spill with paper towels or other absorbent material containing disinfectant.
  2. Cover the area with disinfectant-soaked towels and carefully pour disinfectant around the spill. Avoid enlarging the contaminated area. Use more concentrated disinfectant as it is diluted by the spill. Allow at least 20 minutes contact time.
  3. Remove broken glassware with forceps or a broom and dustpan, and dispose in sharps container. Do not pick up any contaminated sharp object with your hands.
  4. Wipe surrounding areas where the spill may have splashed with disinfectant.
  5. Wipe equipment with 1:10 bleach followed by water then 70% alcohol.
  6. After spill has been absorbed, remove paper towels and any other absorbent material and dispose in biohazard bags.
  7. Soak up the disinfectant and spill and place the materials, along with all contaminated protective clothing, into a biohazard bag. Wipe the area with fresh disinfectant. Decontaminate the utility gloves. Wash hands with disinfectant or soap.
• Reopen area to general use only after spill cleanup and decontamination are complete.
• Inform all personnel and laboratory supervisor about the spill and successful cleanup as soon as possible.

21.4.1 Spills in a Biological Safety Cabinet

The following procedures are to be followed carefully:

• Wear a lab coat, safety goggles and gloves during cleanup.
• Allow cabinet to run during cleanup.
• Spray or wipe cabinet walls, work surfaces, and equipment with disinfectant. If necessary, flood work surface, as well as drain pans below the work surface, with disinfectant. Allow at least 20 minutes contact time.
• Soak up the disinfectant and spill with disposable paper towels and drain catch basin into a container. Lift front exhaust grill and tray and wipe all surfaces. Ensure that no paper towels or solid debris are blown into the area below the tile grill.
• Wipe up spillage and disinfectant with disposable paper towels.
• Wipe the walls, work surface and any equipment in the cabinet with a disinfectant soaked paper towel.
• Discard contaminated disposable materials in biohazard bag(s).
• Place contaminated reusable items in biohazard bags, or heat resistant pans or containers with lids before autoclaving and further cleanup.
• Expose non-autoclavable materials to disinfectant, 10 minutes contact time, before removal from the BSC.
• Remove protective clothing used during cleanup and place in a biohazard bag for decontamination.
• Decontaminate utility gloves and wash your hands.
• Run cabinet at least 10 minutes after cleanup and before resuming work.
• The Biosafety Office should be notified if tile spill overflows into the interior of the cabinet. It may be necessary to do a more extensive decontamination of the cabinet (e.g., formaldehyde decontamination).
• Inform all users of the BSC as well as the laboratory supervisor about the spill and successful cleanup as soon as possible.

21.4.2 Spills Inside a Centrifuge

If a centrifuge tube breaks while the centrifuge is running, turn off the motor. Allow the machine to be at rest for 30 minutes before opening. Or, if breakage is discovered after the machine has stopped, re-close the lid immediately and allow the unit to be at rest for 30 minutes.

• Unplug centrifuge before initiating cleanup.
• Don strong, thick rubber gloves and other PPE before proceeding with cleanup.
• Flood centrifuge bowl with a germicidal disinfectant. Place paper towels soaked in a disinfectant over the entire spill area. Allow 20 minute contact time.
• Use mechanical means (such as forceps) to remove broken tubes and glass fragments. Place them in a sharps container for autoclaving and disposal as infectious waste in accordance with the waste disposal regulations. (See Section 19.)
• Remove buckets, trunnions and rotor, and place in disinfectant for 24 hours or autoclave. Place paper towels soaked in a disinfectant over the equipment if transport is necessary.
• Unbroken, capped tubes may be placed in disinfectant and recovered after 20 minute contact time or autoclaved.
• Use mechanical means to remove remaining disinfectant soaked materials from centrifuge bowl and discard as infectious waste in accordance with regulations. (See Section 19.)
• Place paper towels soaked in a disinfectant in the centrifuge bowl, and allow it to soak overnight. Wipe down again with disinfectant, wash with water and dry. Discard disinfectant soaked materials as infectious waste.
• Remove protective clothing used during cleanup and place in a biohazard bag for autoclaving. Wash hands whenever gloves are removed.

21.5 Spills of Biohazardous Radioactive Material

A biohazardous spill involving radioactive material requires emergency procedures which are different from the procedures used for either material alone. In general, the biological component is inactivated with disinfectant and then the spill is handled as a radioactive spill.

Before any cleanup, consider the type of radionuclide, characteristics of the microorganism, and the volume of the spill. Spills involving ¹⁴C or ³H represent no external hazard to personnel. However, more energetic gamma or beta emitters may require hand and body protection. Do not autoclave any radioactive materials without the approval of the Radiation Safety Office.

Do not use bleach to decontaminate any biohazardous spill containing ¹²⁵I.

21.5.1 Emergency Procedures

• Avoid inhaling airborne material while quickly leaving the room.
• Notify others to leave. Close the door and post a warning sign.
• Remove contaminated clothing, turn exposed area inward, and place in a biohazard bag. Label the biohazard bag as radioactive if applicable.
• Wash all exposed skin with disinfectant, followed by a three-minute water rinse.
• Inform the supervisor and Radiation Safety Officer of the spill.
• Monitor all exposed personnel for radiation.
• If assistance is needed in handling the microorganism, contact the Biosafety Officer.

21.5.2 Cleanup of Biohazardous Radioactive Material

• Allow aerosols to settle for at least 30 minutes before reentering the laboratory.
• Assemble cleanup materials (disinfectant, autoclavable containers, forceps, towels, sponges), and confirm with the Radiation Safety Office that it is safe to enter the lab.
• Put on protective clothing (gown, appropriate mask, gloves, and shoe covers).
• Cover the area with disinfectant-soaked towels, and carefully pour disinfectant around the spill. Avoid enlarging the contaminated area. Use more concentrated disinfectant as it is diluted by the spill. Allow at least 20 minutes contact time.
• Handle any sharp objects with forceps. Wipe surrounding areas where the spill may have splashed with disinfectant.
• Soak up the disinfectant and spill, and place the materials, along with all contaminated protective clothing, into a disposable solid plastic container lined with a biohazard bag, for ultimate disposal as radiation waste. Do not autoclave the waste unless this action is approved by the Radiation Safety Officer. If it cannot be autoclaved, add additional disinfectant to ensure decontamination of all the materials.
• Wash hands and exposed skin areas with disinfectant, and monitor personnel and spill area for residual radioactive contamination. If skin contamination is detected, wash with clean water for three minutes. If spill area has residual activity, determine if it is fixed or removable and handle it accordingly.
• Label everything that is radioactive, including the waste.

21.6 Spills Outside the Laboratory, During Transport in the Institute

Always transport biohazardous materials in an unbreakable well-sealed primary container placed inside a leak-proof, closed and unbreakable secondary container, labeled with the biohazard symbol (plastic cooler, bio-specimen pack, etc).

Should a spill of BL2 material occur in the public area, do not attempt to clean up the spill without the proper personal protective equipment and spill cleanup material.

As an interim measure, wear gloves and place paper towels, preferably soaked in disinfectant, directly on spilled materials to prevent spread of contamination. To assure adequate contact, surround the spill with disinfectant, if available, taking care to minimize aerosols.

Recommended Containment Levels For Infectious Agents