Moffitt, A. D., Brignolo, L. L., Ardeshir, A. et al. 2019. The role of emotional contagion in the distress exhibited by grouped mice exposed to CO2. JAALAS 58(4), 430-437.

The 2013 AVMA Guidelines for the Euthanasia of Animals recommends a chamber volume displacement rate of 10% to 30% per minute (v/min) when euthanizing small laboratory rodents with CO2. Group euthanasia of mice is a common practice, and grouping strangers is often avoided to minimize distress; however, emotional contagion, which occurs between familiar animals but not strangers, has not been studied in the context of group CO2 euthanasia. This study examined cagemate- and stranger-grouped mice exposed to 10%, 30%, or 50% v/min CO2 to determine whether emotional contagion plays a role in this context and whether that role is influenced by CO2 flow rate. Videos of adult male C57BL/6J mice exposed to different CO2 flow rates were scored for durations of dyspnea, ataxia, and consciousness as well as the numbers of face pawing and jump behaviors. Blood was collected at time of unconsciousness and assayed for ACTH. Cagemates experienced significantly longer durations of conscious dyspnea and ataxia with 10% v/min CO2 compared with 30% and 50% v/min. Similarly, strangers experienced significantly longer duration of conscious dyspnea with 10% v/min CO2 compared with 30% and 50% v/min and significantly longer duration of ataxia with 10% compared with 50% v/min. Cagemates showed significantly more jumps with 10% v/min CO2 compared with 30% and 50% v/min, whereas jumping was unaffected by CO2 flow rate in strangers. We conclude that more potential for distress exists when cagemate and stranger mice are exposed to a 10% v/min CO2 flow rate and that emotional contagion may contribute to distress in cagemates at this flow rate. Therefore, we propose that 30% v/min CO2 should be used for euthanasia of mice, and that 50% v/min should also be considered humane.