Different stunning methods (carbon dioxide [CO2] and hypothermia) used in industrial fish processing were compared with asphyxia in air, which is used in traditional fishing, to evaluate the effects of these methods on the stress responses and the meat quality of the Amazon hybrid surubim, Pseudoplatystoma fasciatum female × Leiarius marmoratus male. After the application, blood and behavioral indicators were evaluated. Following death by gill cutting, the fish were gutted, and analyses of muscle pH, rigor mortis, the exudation of the meat, and coloration were performed. Significantly higher levels of cortisol and glucose were exhibited by the fish subjected to asphyxia than by the fish stunned by either CO2 or hypothermia. The fish subjected to asphyxia displayed decreasing muscle pH during the first 2 h after death and showed higher rates of rigor mortis after 3 h than the fish that were stunned by the other two methods. The asphyxia is a practice that exposes fish to suffering by causing an increase in stress responses, which affects meat quality. The immersion of fish in water and ice (hypothermia) for 5 min was more effective stunning method than the use of CO2, resulting in higher loss of sensibility and greater welfare.