Rat aversion to isoflurane versus carbon dioxide

Some experts suggest that sedation of laboratory rodents with isoflurane before euthanasia with carbon dioxide (CO₂) is a humane alternative to euthanasia with CO₂ alone, but little research has compared aversion with these agents. Albino rats were tested in a light–dark box where they had the choice between remaining in a dark compartment filling with isoflurane or CO₂, or escaping to a lit compartment. Experiment 1 validated the procedure by confirming that rats responded to agent and light intensity. In experiment 2, 9/16 and 0/16 rats remained in the dark compartment until recumbent when initially exposed to isoflurane and CO₂, respectively. In experiment 3, more rats remained in the dark compartment until recumbent during initial (10/16) versus re-exposure (1/16) to isoflurane. These results indicate that initial exposure to CO₂ is more aversive than isoflurane, and that re-exposure to isoflurane is more aversive than initial exposure. We conclude that sedation with isoflurane is a refinement over euthanasia with CO₂ alone for rats that have not been previously exposed to inhalant anaesthetics.