The distress experienced by animals during the induction of unconsciousness remains one of the most important and yet overlooked aspects of effective methods of anaesthesia and euthanasia. Here we show that considerable differences exist in the aversive responses elicited by 12 common methods of inhalational anaesthesia and euthanasia in laboratory rats and mice. Carbon dioxide, either alone or in combination with oxygen or argon, was found to be highly aversive to both species. The least aversive agents were halothane in rats and enflurane in mice. Exposing these animals to carbon dioxide in any form, either for anaesthesia or for euthanasia, is likely to cause considerable pain and distress and is therefore unacceptable when efficient and more humane alternatives are readily available.