Safe Work Practices: Biosafety

Upon beginning any work with infectious or dangerous materials, the following steps should be taken to preserve safety and minimize the likelihood of serious injury or death. This document references:

  • “Prudent Practices in the Laboratory” – National Research Council
  • Biosafety in Microbiological and Biomedical Laboratories (BMBL 5th ed.) U.S. HHS.

*Note: If a security access plan is required for your lab, there may be special handling instructions or procedures which must be added to this document. **

Personal Habits:

  • Do not eat, drink, smoke, chew gum, apply cosmetics or remove/insert contact lenses while in the lab
  • Do not store food or beverages in the lab or chemical refrigerator and must be labeled “Not for Human Consumption”
  • These items must be kept in a separate area from the lab and the refrigerator must be labeled as “Items for Human Consumption Only”
  • Food containers of any kind are not fit for chemical storage and are prohibited
  • Do Not mouth pipette
  • Wash hands before leaving the lab or after handling contaminated materials
  • Never bypass engineering/mechanical controls or barriers.
  • Avoid open-toed shoes in the lab
  • Hair should be up kept and away from hazards
  • Proper PPE should be worn to protect clothing, body and reduce risk of injury.
  • No animals, plants, candles, toys or minors are permitted in the lab.

Safe Practices:

  • All materials and equipment should be obtained, cleaned and prepared before the start of any work.
  • Do Not use chipped or cracked glassware
  • Never work in a lab alone or unattended and only with authorization from a professor or PI
  • When working with hazardous materials, always have a second person present
  • Be familiar with emergency procedures specific to your area and duties
  • Keep the lab neat and clean at all times
  • Restore materials to their proper containment areas when finished or at the end of the lab duration
  • Be familiar with decontamination procedures specific to your area and work practices
  • Decontaminate as needed
  • All procedures should be performed at all times necessary to minimize the formation of aerosols
  • Always wear the proper PPE, wear it correctly and remove it correctly
  • Generally, contact lenses should not be used in the lab without approval and proper PPE
  • Familiarize yourself with the Chemical Hygiene Plan (CHP)
  • A pest management system (Vector Control) is available to prevent pests in the lab.

Aerosol Formation Procedures

The formation of aerosols of infectious agents can greatly increase the chances of infection and contamination. Great care should be taken to reduce the risk of generating aerosols in the lab.

All procedures that involve the following must be conducted in a comparable Biosafety Cabinet certified for use with that agent.

Possible aerosol creation,

High concentrations; and

Large volumes,

Innoculation-Loop Manipulations

  • Cub-culturing and streaking procedures
  • Flaming the loop
  • Cooling the loop in culture media

Pipetting

  • Mixing microbial suspensions
  • Spills on hard surfaces

Needle and Syringe Manipulation

  • Expelling air
  • Withdrawal of a needle from a stopper
  • Injecting animals

Centrifuging of Infectious Materials

  • Can be performed in open lab using sealed rotor heads or centrifuge safety cups.
  • Sealed rotor heads are an acceptable method of transfer within the lab. (see transfers)

Hood Systems

  • Use the fume hood or Biosafety cabinet if working with respective items
  • Know when to use one over the other
  • Never use biohazard material in a fume hood.
  • Only use Biosafety, infectious agents or bio-hazardous materials in a Biosafety cabinet (BSC).
  • A flag should be attached to the edge of the hood sash to serve as a visual verification that the hood is running.
  • Clean spills in a fume hood immediately
  • Never store chemicals or containers inside the fume hood or BSC
  • Hoods are for temporary containment only
  • Never stick your head or body into the hood system
  • Never bypass the sash or shield
  • Always use the hood system with the sash at the proper operating level (indicated by arrows)
  • Hands or arms should only enter the hood system once proper PPE is dawned.
  • Allow airflow to stabilize after placing hands/arms inside the BSC (~1min)
  • BSC’s must be decontaminated before and after each use.
  • Waste should be autoclaved and disposed of properly
  • If decontamination is not possible inside the BSC, place item in a sealed container to transport to autoclave
  • Work at least 4 inches from the inside edge of the front grille
  • All items should be placed as far to the rear of the BSC as possible to maximize ventilation

Sharps:

  • A sharps injury log should be implemented in the lab. A sample log is available in the appendix.
  • Sharps must be disposed of in sharps containers which should be:

Puncture Resistant

Conveniently Located

Inside the Biosafety Cabinet

  • Any use of needles with any animals, infectious materials or recombinant DNA must conform to BMBL and NIH guidelines
  • Sharps should be avoided if there is another viable alternative (Syringes, glass etc.)
  • Use integral needle/syringes or needle-locking syringes only
  • Never Re-Cap a syringe
  • Ensure needles are not bent, sheared, recapped or removed from the syringe before disposal
  • Broken glass should not be handled directly
  • Clean broken glass with a broom, dust-pan, tongs or forceps.

Decontamination:

Autoclave

  • Autoclaved materials must be placed inside an autoclave red bag or in a pan. Never overfill
  • Autoclave tape may be used in certain situations
  • Add between 250-500ml water to the bag or pan for steam to generate and circulate
  • A spore strip can be used to verify successful treatment, shown by a color change.
  • Autoclave must reach at minimum 121°C (250° F) for at least 30min at 15PSI
  • After autoclave, material is no longer regarded as infectious
  • Must log each use of the autoclave
  • Maintained for 3 years by dept
  • Required by EPA
  • Ensures proper decontamination taking place

Chemicals

  • Bleach 1:100 for 10-30 minutes is sufficient (1%)
  • Bleach 1:10 for 5 minutes is sufficient (10%)
  • Bleach solutions should be used in a spray bottle
  • Autoclave is the Primary option to decontaminate. Chemicals are Secondary options
  • Vaporized H2O2 is an acceptable method of decontamination
  • Broad spectrum germicide should be available if working with live organisms.
  • Use 70% isopropyl alcohol in a plastic spray bottle to spray all items entering / leaving the BSC.
  • Biohazard autoclave bags should be available for bio-hazardous materials
  • Autoclave bucket should be available to collect contaminated items

Spills or Accidents:

  • Report all injuries to EH&S
  • Know what you are working with and be familiar with its MSDS in case of a spill
  • Ask, “If this spills what will I need to do?”
  • Be familiar with locations of spill kits for small spills and cleanups if applicable to your area

Waste Management:

  • All responsible individuals MUST be trained by EH&S for this topic
  • All items must be labeled at the time of filling a container using a “Green Form” (See EH&S for details)