Deep body temperature is a critical indicator of heat strain. However, direct measures are often invasive, costly, and difficult to implement in the field. This study assessed the agreement between deep body temperature estimated from heart rate and that measured directly during repeated work bouts while wearing explosive ordnance disposal (EOD) protective clothing and during recovery.Eight males completed three work and recovery periods across two separate days. Work consisted of treadmill walking on a 1% incline at 2.5, 4.0, or 5.5 km/h, in a random order, wearing EOD protective clothing. Ambient temperature and relative humidity were maintained at 24 °C and 50% [Wet bulb globe temperature (WBGT) (20.9 ± 1.2) °C] or 32 °C and 60% [WBGT (29.0 ± 0.2) °C] on the separate days, respectively. Heart rate and gastrointestinal temperature (TGI) were monitored continuously, and deep body temperature was also estimated from heart rate (ECTemp).The overall systematic bias between TGI and ECTemp was 0.01 °C with 95% limits of agreement (LoA) of ±0.64 °C and a root mean square error of 0.32 °C. The average error statistics among participants showed no significant differences in error between the exercise and recovery periods or the environmental conditions. At TGI levels of (37.0-37.5) °C, (37.5-38.0) °C, (38.0-38.5) °C, and > 38.5 °C, the systematic bias and ± 95% LoA were (0.08 ± 0.58) °C, (- 0.02 ± 0.69) °C, (- 0.07 ± 0.63) °C, and (- 0.32 ± 0.56) °C, respectively.The findings demonstrate acceptable validity of the ECTemp up to 38.5 °C. Conducting work within an ECTemp limit of 38.4 °C, in conditions similar to the present study, would protect the majority of personnel from an excessive elevation in deep body temperature (> 39.0 °C).