Biohazardous Spill in a Biological Safety Cabinet

Chemical decontamination procedures should be initiated at once while the Cabinet continues to operate to prevent escape of contaminants from the Cabinet.

1. Spray or wipe walls, work surfaces, and equipment with a decontaminant that is effective against the agent in use. Generally a 1% solution of an iodophor decontaminant (Wescodyne or equivalent) is effective against most viruses, fungi, vegetative bacteria, and most non-encysted ameba. A decontaminant detergent has the advantage of detergent activity, which is important because extraneous organic substances frequently interfere with the reaction between microorganisms and the active agent of the decontaminant. The operator is to wear gloves and lab coat during this procedure.
2. Flood the top work surface tray, and, if a Class II Cabinet, the drain pans and catch basins below the work surface, with a decontaminant and allow to stand 1015 minutes.
3. Remove excess decontaminant from the tray by wiping with a sponge or cloth soaked in a decontaminant. For Class II Cabinets, drain the tray into the Cabinet base, lift out tray and removable exhaust grille work, and wipe off top and bottom (underside) surfaces with a sponge or cloth soaked in a decontaminant. Then replace in position and drain decontaminant from Cabinet base into appropriate container and autoclave according to standard procedures. Gloves, cloth, or sponge should be discarded in an autoclave pan and autoclaved.

Biohazardous Spill Outside a Biological Safety Cabinet

1. If you have spilled a risk group 1 agent or a small (unconcentrated) amount of a class 2 agent than remove any contaminated clothing, wash, regard and proceed to step 7.
2. If you are working with a risk group 3 agent, or have spilled more than 100ml of a risk group 2 agent, hold your breath, leave the room immediately, and close the door.
3. Warn others not to enter the contaminated area.
4. Remove and put contaminated garments into a container for autoclaving and thoroughly wash hands and face.
5. Wait 30 minutes before reentering area to allow dissipation of aerosols created by the spill.
6. Put on a longsleeved gown, mask, and rubber gloves before reentering the room. (For high risk agent, a jumpsuit with tightfitting wrists and use of a respirator should be considered.)
7. Pour a decontaminant solution (1% iodophor or 10% hypochlorite are recommended) around the spill and then place towels over the spill. Soak the towels with the decontaminant. To minimize aerosol production, avoid pouring the decontaminant solution directly onto the spill.
8. Let stand 20 minutes to allow an adequate contact time.
9. Using an autoclavable dust pan and squeegee, transfer all contaminated materials (paper towels, glass, liquid, gloves, etc.) into a deep autoclave pan. Cover the pan with aluminum foil or other suitable cover and autoclave according to standard directions.
10. The dust pan and squeegee should be placed in an autoclave bag and autoclaved according to standard directions. Contact of reusable items with non-autoclavable plastic bags should be avoided separation of the plastic after autoclaving can be very difficult.

Radioactive Biohazardous Spill Outside A Biological Safety Cabinet

In the event that a biohazardous spill also includes radioactive material, the cleanup procedure will have to be modified. The biological component of the spill will have to be inactivated prior to disposal of the radioactive waste. Call the Radiation Protection at extension 6626 for instruction and assistance.

Risk Group 1 Agents or small unconcentrated (<100ml)>

1. Warn others not to enter the contaminated area.
2. Remove protective clothing (lab coat, gloves, etc.) and isolate in a plastic bag or appropriate container.
3. Monitor yourself for radioactive contamination. If contaminated decontaminate yourself and resurvey.
4. Thoroughly wash your hands and face.
5. Monitor the removed protective clothing for radioactive contamination. If positive, isolate this waste and hold for disposal by the Radiation Protection Office.
6. Pour a decontaminant solution (1% iodophor or 10% hypochlorite are recommended) around the spill and then place towels over the spill. Soak the towels with the decontaminant. To minimize aerosolization, avoid pouring the decontaminant solution directly onto the spill.
7. Let stand 20 minutes to allow an adequate contact time.
8. Using an autoclavable dust pan and squeegee, transfer all contaminated materials (paper towels, glass, liquid, gloves, etc.) into a plastic bag. Place the bag in the appropriate Radiation waste container.
9. The dust pan and squeegee should be monitored for radioactive contamination. Decontaminate and resurvey as necessary.
10. Contact the Radiation Protection at extension 6626 to report the spill.

Risk Group 3 Agents or spills of Risk Group 2 Agents greater than 100 milliliters.

1. If you spill a Class 3 agent or have spilled more than 100ml of a class 2 agent, hold your breath, leave the room immediately, and close the door.
2. Warn others not to enter the contaminated area.
3. Remove protective clothing (lab coat, gloves, etc.) and isolate in a plastic bag or appropriate container.
4. Monitor yourself for radioactive contamination. If contaminated decontaminate yourself and re-survey.
5. Thoroughly wash your hands and face.
6. Monitor the removed protective clothing for radioactive contamination. If positive, isolate this waste and hold for disposal by Radiation Protection.
7. Prior to reentering the laboratory or spill area, wait 30 minutes to allow dissipation of aerosols created by the spill.
8. Before cleaning the spill area contact Radiation Protection at extension 6626 for assistance. If the spill occurs after hours or on weekends, activate the Emergency Response System by dialing 7911

Chemical Biohazardous Spill Outside of a Biological Safety Cabinet

1. Determine prior to starting your research: which chemical decontaminant(s) is compatible with the chemical(s) that may become biologically contaminated and if the contaminated chemical(s) can be autoclaved. Contact the Industrial Hygienist at extension 6218 for assistance.
2. If you are working with a Risk Group 3 agent, or have spilled more than 100ml of a Risk Group 2 agent, hold your breath, leave the room immediately, and close the door.
1. If you have spilled a Risk Group 1 agent or a small amount of a Risk Group 2 agent then proceed to step 3.
3. Warn others not to enter the contaminated area.
4. Remove and put in a container contaminated garments for decontamination or autoclaving and thoroughly wash hands and face. If garments are chemically contaminated, autoclaving may not be advisable, consult with an industrial hygienist .
5. If you have had to evacuate the laboratory, wait 30 minutes to allow dissipation of aerosols created by the spill.
6. Consult your laboratory's Chemical Hygiene Plan for chemical spill procedures. If the chemical(s) in the spill present a greater hazard then the biological agent(s) proceed with chemical decontamination first.
7. Put on a longsleeved gown, mask, and rubber gloves before reentering the room. For high risk agent or hazardous chemical, a jumpsuit with tightfitting wrists and use of a respirator (consult with the Industrial Hygienist ) should be considered.
8. Use a decontaminant that is compatible with the chemical(s) in the spill. Pour the decontaminant solution around the spill and then place towels over the spill. Soak the towels with the decontaminant. To minimize aerosol production, avoid pouring the decontaminant solution directly onto the spill.
9. Let stand 20 minutes to allow an adequate contact time.
10. If the chemical(s) are compatible with autoclaving, use an autoclavable dust pan and squeegee to transfer all contaminated materials (paper towels, absorbent, glass, liquid, gloves, dust pan, squeegee, etc.) into an autoclavable pan. Cover the pan with aluminum foil or other suitable cover and autoclave according to standard directions. Depending upon the chemicals involved the now sterile material may have to be disposed of via Waste Management's chemical waste pickup service.
11. If the chemical(s) are not autoclavable (or if you do not know), then transfer the decontaminated, contaminated materials into a screw cap container and call Waste Management at extension 5877 for chemical waste pickup
12. Contact of reusable items with nonautoclavable plastic bags should be avoided separation of the plastic after autoclaving can be very difficult.