AbstractThe Largemouth Bass Micropterus salmoides, a popular sport fish, is subjected to multiple sublethal stressors during angling, including high water temperature, exercise, handling, live‐well retention, and weigh‐in procedures. Combined effects of ambient and live‐well temperatures on the stress response and recovery from angling‐induced exercise have not been tested in conditions similar to those encountered in tournaments. Therefore, we assessed the effects of ambient temperature (17, 25, and 33°C) and live‐well temperature differential (−4, 0, and +4°C) on the physiological stress response of Largemouth Bass (mean length = 331 mm) at rest, following a simulated angling stressor, and throughout 8 h of recovery in live wells. Stress variables were measured in whole blood (hematocrit, hemoglobin, pH, partial pressure of oxygen [pO2], partial pressure of carbon dioxide [pCO2], Na+, K+, Ca2+, Cl−, and leukocytes) and plasma (cortisol, glucose, lactate, and osmolality). Fish acclimated to 17°C showed the greatest cortisol response after the chasing stressor; however, higher levels of glucose, lactate, pCO2, K+, and monocyte percentage were found at 33°C, and blood pH, Cl−, and lymphocyte percentage were lower at 33°C than at 17°C. When live‐well temperature was manipulated, cortisol levels were highest in fish subjected to the coldest conditions (acclimated to 17°C and retained in 13°C and 17°C live wells) and the warmest condition (acclimated to 33°C and retained in 37°C live wells). However, all fish subjected to the colder extremes survived, whereas 100% mortality occurred in the warmest condition. Besides cortisol, indicators of stress were less pronounced at colder temperatures. Glucose, lactate, and notably K+ concentrations were highest in 37°C live wells, and blood pH, Ca2+, Na+, and Cl− were lowest. Low blood lymphocytes and high monocytes at the warmest conditions indicate reduced immunocompetence or inflammation. Mortality at high temperature may result from exhaustion of aerobic and anaerobic energy sources, failure to recover from metabolic acidosis, and an inability to regain ionic balance.