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S.O.P. Detailing Spillage Cleanup, Inactivation, and Record Keeping

Preamble

In order to prepare for spillage of GMM materials, a spill kit is to be maintained in the lab at all times. The kit is for use only as a designated spill kit and not for general purpose use. The kit must never be used outside the lab. The kit should consist of cleaning equipment and cleaning agents, and should minimise use of brushes or cloths, favouring sterilisable and disposible absorbants instead. A mop can be used for final cleanup after collection and inactivation of the spill residue.

Aims

To absorb, remove and sterilise spillage waste, and to sterilise effectively the spill site, without leaving viable biological residues at the site of the spill or on spill-kit equipment.

Protocols

Protocol 1: Maintenance of Spill Kit

The spill-kit should consist of the following, in sufficient quantity where relevant to address any foreseeable spillage size. If spillage hazards exceed this amount (as may occur during bioreactor scale-up), the spill kit must be increased in handling capacity to meet the new potential spill volume.

  • Disposable gloves.
  • An absorbant material, preferably in pellet or wafer form, that can be used to absorb primary spillage for removal. i.e. Wood-chip, spill-granules, disposible kitchen tissue.
  • A strong disinfectant agent for destruction of viable vegetative cells. i.e. Dettol
  • A strong oxygen bleaching agent for subsequent destruction of sporulated cells. i.e. Sodium percarbonate, toilet bleach.
  • A mop and bucket for removal of bleach from decontaminated area.
  • A bucket for transitory storage of contaminated absorbants.
  • Autoclavable plastic bags for storage of absorbants.

Protocol 2: Spill Cleanup Procedure

  1. In case of spillage, absorbants should be immediately placed around the spill to prevent further spread, and then on the spill itself to soak the liquid. Gloves should be worn as soon as the immediate spill is contained.
  2. Soaked absorbants should be placed in autoclavable bags and removed immediately to the autoclave via the transitory waste-bucket. If the autoclave is mid-cycle, waste should be stored in the transitory bucket until it can be sterilised.
  3. Once the liquid has been absorbed and removed to bags, the spill area should be doused in primary disinfectant, and left to inactivate cells for as long as required by the product plus a marginal time of five minutes.
  4. The primary disinfectant should be absorbed and absorbants kept in autoclavable bags, for inactivation along with spill waste.
  5. Secondary oxygen-based bleach should be applied to the area if the spillage contained endospore-forming bacteria, to inactivate spores. The prefferred agent is sodium percarbonate. This should be allowed an hour of contact time in which to work. It should then be absorbed and disposed of in like manner to the primary disinfectant.
  6. The area should be washed with a mop and bucket to remove bleach and then dried conventionally.
  7. Containers used in the spill kit, such as the transitory waste-bucket, must likewise be sterilised using disinfectant and then bleach treatment after use.
  8. A record entry must be made in the “Inactivation Records” book detailing the spill nature and outcome, including the results of the verification procedure, below.

Verification

Swabbing and culture of surfaces, with selective agents where appropriate and PCR in all cases, should be performed to verify destruction of viable cells.

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