An animal euthanasia device includes a chamber with an opening through which a partitioned cage may be wheeled. A pivotally attached door seals the opening using fixed edge seals and an inflatable door threshold seal. A latch secures the door, once closed. After a sensor verifies door closure, an electro-magnetic lock prevents inadvertent door opening during the process, which may be automated using a programmable logic controller (PLC). The PLC coordinates actuation of a fan and damper within an air exhaust opening, a fan and a damper within an exhaust gas opening, and a damper within a fresh air inlet opening, and also triggers a solenoid valve that controls flow of euthanizing gas into the chamber. A keyed selector switch controls power to the device to initiate the automated process, which is monitored using oxygen sensors, and CO2 flow and pressure sensors, and is reported on a human machine interface.