Striving for humane deaths for laboratory mice: hypobaric hypoxia provides a potential alternative to carbon dioxide exposure

Killing is often an unavoidable and necessary procedure for laboratory mice involved in scientific research, and providing a humane death is vital for public acceptance. Exposure to carbon dioxide (CO2) gas is the most widely used methodology despite well proven welfare concerns. Consequently, the continued use of CO2 and its globally permitted status in legislation and guidelines presents an ethical dilemma for users. We investigated whether killing with hypobaric hypoxia via gradual decompression was associated with better welfare outcomes for killing laboratory mice. We compared the spontaneous behaviour of mice exposed to CO2, decompression or sham conditions, and used analgesic or anxiolytic interventions to determine their relative welfare impact. Gradual decompression resulted in longer times to unconsciousness and death and the pharmacological interventions support the notion of a minimally negative animal experience, while providing further evidence for pain and anxiety associated with exposure to CO2. Decompression resulted in moderate ear haemorrhage, but our welfare assessment suggests this may happen when mice are unconscious. Hence, gradual decompression could be the basis of significant refinement for killing laboratory mice. Future work should corroborate behaviour with neurobiological markers of loss of consciousness to verify the conscious phase of concern for animal welfare

Physiological and behavioral responses of poultry exposed to gas-filled high expansion foam

Disease control measures require poultry to be killed on farms to minimize the risk of disease being transmitted to other poultry and, in some cases, to protect public health. We assessed the welfare implications for poultry of the use of high-expansion gas-filled foam as a potentially humane, emergency killing method. In laboratory trials, broiler chickens, adult laying hens, ducks, and turkeys were exposed to air-, N2-, or CO2-filled high expansion foam (expansion ratio 300:1) under standardized conditions. Birds were equipped with sensors to measure cardiac and brain activity, and measurements of oxygen concentration in the foam were carried out. Initial behavioral responses to foam were not pronounced but included headshakes and brief bouts of wing flapping. Both N2- and CO2-filled foam rapidly induced ataxia/loss of posture and vigorous wing flapping in all species, characteristic of anoxic death. Immersion in air-filled, high expansion foam had little effect on physiology or behavior. Physiological responses to both N2- and CO2-filled foam were characterized by a pronounced bradyarrythymia and a series of consistent changes in the appearance of the electroencephalogram. These were used to determine an unequivocal time to loss of consciousness in relation to submersion. Mean time to loss of consciousness was 30 s in hens and 18 s in broilers exposed to N2-filled foam, and 16 s in broilers, 1 s in ducks, and 15 s in turkeys exposed to CO2-filled foam. Euthanasia achieved with anoxic foam was particularly rapid, which is explained by the very low oxygen concentrations (below 1%) inside the foam. Physiological observations and postmortem examination showed that the mode of action of high expansion, gas-filled foam is anoxia, not occlusion of the airway. These trials provide proof-of-principle that submersion in gas-filled, high expansion foam provides a rapid and highly effective method of euthanasia, which may have potential to provide humane emergency killing or routine depopulation