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The purpose of this study was to examine the effect of heat acclimation (HA) on thirst levels, sweat rate, and percentage of body mass loss (%BML), and changes in fluid intake factors throughout HA induction. Twenty-eight male endurance athletes (mean ± SD; age, 35 ± 12 years; body mass, 73.0 ± 8.9 kg; maximal oxygen consumption, 57.4 ± 6.8 ml·kg−1·min−1) completed 60 min of exercise in a euhydrated state at 58.9 ± 2.3% velocity of maximal oxygen consumption in the heat (ambient temperature, 35.0 ± 1.3 °C; relative humidity, 48.0 ± 1.3%) prior to and following HA where thirst levels, sweat rate, and %BML were measured. Then, participants performed 5 days of HA while held at hyperthermia (38.50–39.75 °C) for 60 min with fluid provided ad libitum. Sweat volume, %BML, thirst levels, and fluid intake were measured for each session. Thirst levels were significantly lower following HA (pre, 4 ± 1; post, 3 ± 1, p p = .039) and %BML (pre, 2.66 ± 0.53%; post, 2.98 ± 0.83%, p = .049) were significantly greater following HA. During HA, thirst levels decreased (Day 1, 4 ± 1; Day 2, 3 ± 2; Day 3, 3 ± 2; Day 4, 3 ± 1; Day 5, 3 ± 1; p p = .010) and fluid intake (Day 1, 1.20 ± 0.45 L; Day 2, 1.52 ± 0.58 L; Day 3, 1.69 ± 0.63 L; Day 4, 1.65 ± 0.58 L; Day 5, 1.74 ± 0.51 L; p