Workers should aim to fully replace fluid losses during work in the heat to maintain work productivity and minimize risk of heat illnesses, however, fluid balance of females working during consecutive days in the heat is unknown. The purpose of this study was to test the hypothesis that females would maintain fluid balance during work across two consecutive workdays. Eight healthy females (age: 29±5, body mass: 75.3±20.1, body fat: 23±8%) completed two consecutive days (Day 1, Day 2) of heavy intensity work in a hot environment (35.9±1.2°C, 24±6% relative humidity). Work consisted of repeated 45/15 work/rest cycles of treadmill walking or cycling at 398±7 W of metabolic heat production and seated rest for 4 h. Every 20 min, participants were provided 237 mL of a sport drink to consume ad libitum. Urine flow rate (UFR) was measured before work. Urine specific gravity (USG) and nude body mass were measured before and after work. Fluid volume consumed and thirst (9-point Likert scale, 1=not thirsty at all, 9=very, very thirsty) were measured during work. Two-way repeated measures analysis of variance and paired samples t-tests were used to compare differences. Significance was set at p≤0.05. Percent body mass loss was not different between Day 1 (0.69±0.82%) and Day 2 (0.96±2.25%, p=0.935). Similarly, sweat rate was not different between Day 1 (0.57±0.12 L/h) and Day 2 (0.65±0.21 L/h, p=0.312). Participants consumed less fluid on Day 1 (64±21%, 25±10 mL/kg) compared to Day 2 (74±20%, p=0.026; 29±10 mL/kg, p=0.025). Participants consumed 68±19% and 77±20% (p=0.040) of fluid offered during work and 55±27% and 66±22% (p=0.173) of fluid offered during rest on Day 1 and Day 2, respectively. Resting thirst (Day 1: 2.1±1.1, Day 2: 1.6±0.5, p=0.096), change in thirst (Day 1: -0.5±0.9, Day 2: +0.3±1.3, p=0.111) and maximum thirst (Day 1: 3.6±1.1, Day 2: 3.6±1.4, p=1.000) were not different between days. USG was not different (p=0.839) across days before (Day 1: 1.009±0.010, Day 2: 1.009±0.010) or after (Day 1: 1.012±1.010, Day 2: 1.012±0.010) work, respectively. UFR was not different between days (Day 1: 2.2±1.4 mL/min, Day 2: 2.0±1.2 mL/min, p=0.836). Females completing heavy intensity work in heat maintained fluid balance across two consecutive days when fluids were readily accessible. Ad libitum fluid consumption may be suffcient to adequately replace fluid losses in females across consecutive days of work in heat. This project was funded by the National Institute for Occupational Safety and Health (1K01OH012016-01A1). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.