Heart rate variability (HRV) has shown potential as a tool for monitoring thermal strain, but there is limited data to support its efficacy in older adults during prolonged heat exposures. We compared HRV between young (19-31 years, n=20) and older (61-78 years, n=39) adults during 9 hours of heat exposure (40°C, 9% RH). We also explored whether heart rate (HR) and/or HRV could be used to distinguish older adults who achieved elevated thermal strain, defined as either 1) an increase in core temperature >1.0°C (occurring in 39% [15/39]) or 2) a reduction in systolic blood pressure >10 mm Hg (occurring in 67% [26/39]). Percentage of age-predicted maximal HR and percentage of heart rate reserve (HRR) were higher, whereas standard deviation of normal RR intervals (SDNN), the square root of the mean of squared differences between successive RR intervals (RMSSD), high frequency power (HF), and cardiac vagal index (CVI) were lower in older compared to young adults (P≤0.004) during heat exposure. In older adults, increases in core temperature were correlated with percentage of age-predicted maximal HR, percentage of HRR, RMSSD, and CVI (P≤0.031), whereas changes in systolic blood pressure were not significantly associated with HR or HRV indices (P≤0.327). Receiver operating characteristic curve analysis indicated that HR and HRV indices had generally poor ability to identify older adults with elevated thermal strain (area under the curve ≤0.65). Age-related differences in HRV, consistent with vagal withdrawal among older adults, remained during daylong heat exposure, but marked heterogeneity of response likely contributed to HRV providing limited discriminatory value in identifying changes in core temperature or blood pressure in older adults.