One of the most critical steps in fish welfare occurs in the slaughter phase, with hormonal, biochemical, osmoregulatory and energetic alterations. These changes can affect the meat quality, with changes in colour and texture of the fillets, protein degradation, and lipid and metabolic oxidation (Daskalova et al.,2016;Zampacavallo et al., 2015).Various methods for stunning and killing fish have been studied, such as electrical current application, cranial percussion, hypothermia, asphyxiation, CO 2 narcosis, anaesthesia, and spiking. Among these methods, stunning by hypothermia has been one of the most used by the fish industry (Zampacavallo et al., 2015; Oliveira Filho, Oliveira, et al., 2015a; Oliveira Filho, 2015b). However, the rapid decline in the body temperature of fish can result in animal discomfort and delayed loss of consciousness Zampacavallo et al., 2015), increasing the plasma cortisol and catecholamines levels (Seth et al., 2013).CO 2 narcosis has various advantages, including the use of watersoluble gas, ease to apply and safe to handle (Rahmanifarah et al.,