Evaluation of Carbon Dioxide Dissipation within a Euthanasia Chamber

CO₂ euthanasia is used widely for small laboratory animals, such as rodents. A common necessity in many animal research facilities is to euthanize mice in sequential batches. We assessed the effects of several variables on the time it took for CO₂ to dissipate within a chamber. Using standard euthanasia time, changes in flow rate were compared between a slow 15% fill rate for 7 min, and a slow 15% followed by a rapid 50% filling for a total of 5 min. Additional variables assessed included the effects of opening the lid after the completion of chamber filling, turning the chamber over after completion of filling, and the use and removal of a cage from within the chamber. For all trials, CO₂ levels in the chambers peaked between 50% and 80%. After the gas was turned off, the concentration of CO₂ dropped to below 10% CO₂ within 2 min, except when the lid was left on the chamber, where concentration levels remained above 10% after 20 min. CO₂ dissipation was significantly faster when the chamber was turned upside down after filling. Significant interaction effects occurred among the factors of cage presence within the chamber, flow rate, and chamber position. Only leaving the lid on the chamber had any practical implication for delaying CO₂ dissipation. We recommend that users allow 2 min for CO₂ to clear from the chamber before subsequent euthanasia procedures, unless the chamber is manipulated to increase the dissipation rate.