Biological Spill Procedure Guide

Biosafety Risk Groups:

Biosafety Level 1 (BSL 1) – Organisms are well-characterized agents not known to cause disease in healthy adult humans and are of minimal potential hazard to laboratory personnel or to the environment. Examples include B. subtilis, E. coli, and L. acidophilus.

Biosafety Level 2 (BSL 2) – Organisms are agents of moderate potential hazard to laboratory personnel and the environment. Examples include Salmonellae, Hepatitis B virus, bloodborne pathogens, and human body fluids – particularly when visibly contaminated with blood.

Biosafety Level 3 (BSL 3) – Organisms are indigenous or exotic agents which may cause serious or potentially lethal disease and present the potential for aerosol transmission. Examples include H5N1 Influenza virus, Bacillus anthracis, Yersinia pestis, Burkholderia, Francisella tularensis, Brucella, Clostridium botulinum, Mycobacterium tuberculosis, Coxiella burnetii, Hantavirus, and West Nile virus.

Biosafety Level 4 (BSL4) – Organisms are highly pathogenic and require handling in special laboratory facilities designed to contain them. Examples include viral hemorrhagic fevers, such as Ebola, Lassa fever, Hantavirus pulmonary syndrome (HPS) and hemorrhagic fever with renal syndrome (HFRS).

Determine the biosafety level before deciding how to clean and/or contain the spill!

In the event of a Biosafety Level 1 spill:

1. Notify laboratory personnel in the area and restrict access to the spill area to prevent the remainder of the laboratory from becoming contaminated.
2. Get the Spill Kit and put on the appropriate personal protective equipment (PPE): lab coat, eye protection, and gloves.
3. Place a paper towel or some other absorbent paper product over the spill.
4. Spray the paper towel or other absorbent with a fresh 10% bleach and water solution or equivalent.
5. Allow the absorbent material and sterilization compound to set on spill for five to fifteen minutes, depending on the size of the spill.
6. During the five to fifteen minute sterilization period, prepare the red biohazard bag by opening the bag and folding it down from the top so that a wide opening is created and contamination of the outside bag surfaces during filling is prevented.
7. When the five to fifteen minute sterilization time is up, put the soaked towels or absorbent material in the biohazard bag, then wipe up any remaining spill residue with clean paper towels and place them in the biohazard bag.
8. Clean the spill area again with the fresh 10% bleach and water solution or equivalent, placing the paper towels in the biohazard bag when finished.
9. Remove gloves, taking care not to touch the outside surfaces of the gloves with your bare hands, then place them in the biohazard bag.
10. Wash hands, thoroughly.
11. Steam sterilize the loosely closed biohazard bag and contents by autoclave, place the sterilized biohazard bag inside a standard black trash bag, and dispose of it as standard waste, or
12. Close and seal the biohazard bag and arrange for EH&S to pick it up and dispose of it.

For a Biosafety Level 2 Spill:

Follow the procedures for a Biosafety Level 1 spill, but take extra precaution if needles or broken glass are involved.

For Biosafety Levels 3 and 4 Spills:

1. Immediately notify the Office of Environmental Health and Safety at (479)575-5448 or dial 911 and give the following accident details:
   • location
   • pathogen or microorganism involved
   • whether or not any individual was affected (contaminated) by the spill
2. Limit access to the area. Do not allow anyone out, and no one other than EH&S or medical personnel in to the area.

MAJOR SPILL: In the event of a spill large enough to result in liquids flowing through the front or rear grilles requiring more extensive decontamination:

1. Decontaminate the surface of all items inside the BSC using a fresh 10% bleach and water solution or approved equivalent while removing them from the cabinet.
2. Pour the 10% bleach and water solution or equivalent onto the BSC work surface and through the grille(s) into the drain pan.
3. Allow twenty to thirty minutes for decontamination to take place. The amount of time varies according to the pathogen or microorganism involved.
4. Wipe the work surface with paper towel(s) or other approved absorbent and place into the biohazard bag.
5. Use clean paper towel(s) moistened with sterile water to wipe down BSC surfaces again so as to remove any residual bleach solution, then place paper towel(s) in the biohazard bag.
6. Empty the contents of the drain pan into a container that has some of the fresh 10% bleach and water solution or equivalent.
7. Attach flexible tubing to the drain valve. Tubing should be long enough to allow the open end to be submerged in the disinfectant inside the container mentioned above. This technique minimizes aerosol generation.
8. Thoroughly rinse the drain pan with water and drain the contents through the tubing. Remove drain tubing.
9. Remove gloves and wash hands.
10. Put on clean gloves and return everything to inside of BSC.

MINOR SPILL: In the event of a small spill within the BSC, handle immediately:

1. Cover the spill with a fresh 10% bleach and water solution or approved equivalent, allow it to sit for five to fifteen minutes, depending on the size of the spill, then wipe it up with a paper towel or other absorbent.
2. Remove the contaminated absorbent paper towel and place it in a biohazard bag inside the BSC.
3. Wipe the surface again with sterile water and clean paper towel(s) to remove any residual bleach, then place the paper towel(s) in the biohazard bag.
4. Wipe up any splatter on items within the cabinet, as well as the cabinet interior, with a paper towel moistened with fresh 10% bleach and water solution or approved equivalent.
5. Remove contaminated gloves and wash hands.
6. Put on clean gloves and return everything to the cabinet.