Disposal Procedures for Transgenic Insects Used in Research

Transgenic Insects Used in Research Biohazardous Waste Collection, Termination and Disposal Guide for UTK/UTIA/CVM Researchers

Per NIH (April 2021), “The NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines) requires the containment and appropriate disposal of waste containing recombinant or synthetic nucleic acid molecules.   

All materials, including insects, nematodes, fish, and other animals containing recombinant or synthetic nucleic acid molecules should be disposed of in accordance with the requirements of the NIH Guidelines.  The requirement is not only that recombinant or synthetic nucleic acid molecule-containing organisms are dead (devitalized). Appendix G-II-A-1-c of the NIH Guidelines specifically states that at Biosafety Level 1 and higher, all contaminated liquid or solid wastes must be decontaminated before disposal. Appropriate mechanisms for the decontamination of solid waste including recombinant animal waste, includes, but is not limited to, incineration or autoclaving.”

The following procedures are appropriate methods of waste decontamination and disposal that meet the expectations of the NIH Guidelines:

1. Onsite autoclave and disposition – Autoclaving will serve as both devitalization and decontamination. If onsite autoclaving will take place with subsequent disposition in the regular trash, a clear autoclave bag must be used.  Red or orange bags or any bag emblazoned with the biohazard symbol must be placed in the red Advantra bins for final disposition.  If insects will be collected in the autoclave bag over time, they must be devitalized first to prevent possible escape.
2. Offsite treatment by the medical waste contractor (Advantra) – Devitalize the materials and deposit them in the bins provided by our medical waste contractor. Do not place live insects in the bins. An acceptable method for devitalizing is to place the insects in a -20 deg C freezer. The insects can be frozen in their primary containers and then placed in the biohazardous waste receptacle.
3. Chemical treatment – Determine the appropriate chemical and the correct dilution to kill the insects. This will serve as devitalization and decontamination.  The container holding the chemical and insect debris should be leak-proof, wide-mouthed, sealable, made of plastic rated to hold the chemicals used, and labeled as “insect waste” with the type of chemical and concentration on the waste collection label.  When ready for disposal, seal the container and ensure all contents are listed on the waste container.  Take the full waste containers to the waste room on assigned collection days.  Alternatively, if the chemical in use is dilute bleach, after fully devitalized, insects can be strained from the dilute bleach and disposed of in the biohazardous waste container.  Follow by sink disposing of the dilute bleach with copious amounts of water.

Note:  Non-transgenic insects may require similar treatment depending on the level of risk and regulatory oversite (e.g., PPQ permit provisions).  Contact EHS Biosafety for additional guidance.