Isoflurane Is Effective for Anesthesia of the Greater Wax Moth (Galleria mellonella) Larvae

Galleria mellonella (wax moth) larvae are increasingly used as model organisms in biomedical research, yet ethical guidelines for their anesthesia and euthanasia are lacking. A review of published papers revealed that most projects involving G. mellonella either did not use any anesthesia or euthanasia, or employed suboptimal methods that may cause pain. This study demonstrates that brief exposure to isoflurane at 2 μL/cm³ provides rapid, long-lasting anesthesia without impairing immune functions of the larvae. Additionally, combining isoflurane anesthesia with subsequent freezing at -20 °C offers an effective euthanasia method. The described anesthesia and euthanasia methods are simple, scalable, humane, and well-suited for laboratory use in G. mellonella, supporting improved welfare standards in invertebrate research.