Biosafety Guide

Biosafety Level 2 Containment Work Practices

Hand Hygiene

• Wash hands upon contact with anything potentially infectious, after work, after removing gloves and before leaving the laboratory
• Gloves are replaced as soon as practical when contaminated, torn, punctured, or when their ability to function as a barrier is compromised

Aerosol Containment

• Perform all procedures in a manner that minimizes splashes, the creation of droplets or aerosols
• Perform procedures that may generate infectious aerosols in a biosafety cabinet
• Use a mechanical pipetting devices at all times

Universal Precautions

• Universal precautions is an approach to infection control in which you treat all human and nonhuman primate blood, tissues and body fluids as if they are potentially infectious for HIV, hepatitis B virus, and other bloodborne pathogens, (Bloodborne Pathogens Standard 29 CFR 1910.1030(b) definitions).
• Implementing universal precautions means that you consistently adhere to
  • Hand hygiene
  • Wearing non-porous personal protective equipment as a physical barrier between you and the potentially infectious material
  • Safe work practices specific to sharps
  • Routine surface disinfection
  • Waste deactivation and disposal

Delineation of Laboratory from Non-laboratory Operations

• Entry ways to work areas, containers of biohazardous waste, refrigerators and freezers used with potentially infectious materials must have labels with the word “BIOHAZARD” and the universal biohazard symbol in orange-red or red with lettering and symbols in a contrasting color.
• Eating, drinking, smoking, applying cosmetics or lip balm, and handling contact lenses are prohibited in work areas
• Do not handle ear buds or cell phones while wearing PPE
• Food and drink must not be stored in laboratory refrigerators, freezers, shelves, cabinets, or bench tops that contain research material(s)
• Transport biological agents between laboratories and campus buildings using rigid, leak-proof, double container systems that are lined with enough absorbent material to absorb liquid leaks

Disinfection

• Disinfect the work area and lab equipment daily and after use
• Disinfectants don't inactivate microorganisms instantaneously, and the disinfectant needs to dwell on surfaces, e.g., gasketed containers, biosafety cabinet interior, liquid handling equipment used with disposable tips/pipettes, lab surfaces, equipment, safety glasses and face shields
• Use an EPA-registered, high level disinfectant for work with potentially infectious and infectious materials

Housekeeping

• Reduce or eliminate the amount of cardboard 
• Take out the recycling and don't allow piles of it to accumulate 
• Organize freezer space to ease access and inventory of materials 

Written Procedures

• Standard Operating Procedures should be established for techniques, e.g., cell culturing or tissue homogenization, and equipment, e.g., fluorescence activated cell sorters or sonicators
• Methods to prevent occupational exposures are specified by the Principal Investigator within the Biological Use Authorization

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Personal Protective Equipment

• Long pants and closed toe shoes are required for entering and occupying a laboratory
• While working with, or being adjacent to hazardous materials or processes, an appropriate lab coat and safety eyewear are also required
• Wear a cuffed lab coat and disposable gloves when working at a biological safety cabinet
• Wear safety glasses, with an accompanying face shield for handling large volumes for work outside of a biological safety cabinet
• The cuffed, pin-striped barrier lab coats are impervious to small volumes of liquids and are used for work with human blood or other infectious liquid handling procedures
• Lab coats must be visibly clean and laundered as needed
• In case of gross contamination with infectious biological materials, lab coats are to be bagged and autoclaved prior to laundering via Mission Linen

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Relocation Guide for Biological Safety

Packing and Transporting Biological Materials across Campus

• Use a rigid, leak-proof, double container system
• Containers must be readily decontaminated, i.e., not Styrofoam or cardboard
• Seal both the inner and outer containers with Parafilm
• Pack items to prevent breakage or leakage
• Line the outer container with enough absorbent material to soak up the volume of liquid
• Do not leave packages unattended
• Label the container with the biohazard symbol and the PI/designee contact information
• Decontaminate the outside of the primary and secondary containers, including any handles, before you exit the lab, so that you can safely doff PPE before exiting
• Bring clean disposable gloves to unpack the container, and carry your lab coat in an unused autoclave bag

Steps to Relocating a Biosafety Cabinet

1. Authorized personnel/researchers empty the cabinet and decontaminate interior work areas
2. An outside vendor, Technical Safety Services, Inc., performs a decontamination procedure on the entire unit
3. UCSB Furniture Services moves the cabinet to the new location, or into surplus
4. The PI’s departmental staff finalizes installation 
5. Technical Safety Services, Inc. recertifies cabinet performance

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Spill Cleanup

A spill kit, consisting of paper towels, tongs, and disinfectant, is ideally located under or next to the sink. Decontamination of spills within an operating biosafety cabinet proceeds immediately. Keep the cabinet blower on.

1. Replace any contaminated personal protective equipment.
2. Obtain or prepare a fresh solution of disinfectant.
3. Remove any contaminated paper towels and place it into the biohazard bag.
4. Cover the area of the spill with new paper towels to prevent aerosols and splashing, and carefully pour disinfectant on the towels, starting from the outside of the spill.
5. Allow for the contact time specified for the disinfectant; 20 minutes’ contact time for freshly diluted 10% bleach (final vol/vol).
6. Dampen a towel with disinfectant and wipe down the cabinet interior and items inside the BSC.
7. Push towels from the edge of the spill to the center and dispose of into a biohazard bag.
8. Use paper towels to absorb any remaining liquid, working from the outside in.
9. Bag the clean-up materials and disposable gloves as solid waste.
10. Change gloves after the work surface is decontaminated and before replacing clean absorbent towels, if used.
11. Wash your hands thoroughly.

For larger spills outside of a biosafety cabinet and depending on the materials involved, you may need to vacate the premises for 30 minutes to allow for aerosols to settle and for a few room air exchanges to take place.

1. Alert others that there has been a biological spill.
2. Remove one glove to open and then close the door behind you, and leave for 30 minutes.
3. Remove PPE. Bag contaminated items such as shoes or lab coat for surface decontamination or autoclaving. Wash your hands.
4. Post a “Spill Cleanup in Progress” sign at the door warning of the spill and advising of the re-entry time.

Spill Cleanup in Progress
Do not enter the lab.
Re-entry for spill cleanup __:___
This sign will be removed afterwards.
Contact:

5. Before entry, put on a fresh lab coat, disposable gloves, and safety glasses.
6. Obtain or make a fresh dilution of 10% household bleach (final vol/vol).
7. Cover the area with paper towels. Pour (do not spray) 10% bleach on absorbent material in a spiral motion from the outside to the inside. Avoid enlarging the contaminated area.
8. Allow for 20 minutes’ contact time.
9. While waiting, wipe down nearby stationary equipment and furniture with towels dampened with disinfectant.
10. Start at the perimeter and work towards the center to remove towels and place in a biohazard bag.
11. Change gloves.
12. Clean the area again with towels dampened with disinfectant.
13. Remove PPE and wash your hands.
14. Inform the lab when cleanup is complete.
15. Contact EHS Biosafety to report the spill and inform your PI.

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