The measurement of body temperature is a critical aspect of assessing the health and reproductive status of dairy cows. The standard method used to estimate this temperature is rectal thermometry. However, this technique has limitations, including disease spread, distress, and or risks of rectal injuries. The current study was undertaken to validate the potential of alternative anatomical sites for temperature measurement using a digital thermometer (DT). The study employed a one-factor experimental design considering the anatomical site as the main factor, with four treatments or factor levels, namely rectal (DTtrectal), inguinal (DTtinguinal), axillary (DTtaxillary), and undertail (DTtundertail) sites. A simple random sampling technique was employed to determine the order of site selection for temperature measurement. In total, 26 adult Holstein Friesian-Boran cows with an average weight of 482 kg were used to conduct this study. Each cow was assessed for all the treatments considered in this study. The temperature measured at different anatomical sites was evaluated. The highest mean temperature was observed for rectal temperature (38.27 ± 0.42℃), while that of mean axillary temperature was the lowest (37.75 ± 0.53℃). The mean temperature readings were significantly affected by the anatomical site. There was no significant difference between mean rectal and inguinal or undertail temperature. There was a significant correlation between the rectal and undertail temperature, while no significant correlation was observed between rectal and inguinal temperature. The equivalence analysis between the rectal and undertail pair revealed a significant bias. This bias suggests that the two anatomical sites cannot be used interchangeably, particularly with digital thermometer application in Holstein Friesian-Boran cows. However, the observed mean undertail temperature and its correlation with rectal temperature indicated that the undertail site still holds promise as an alternative site for temperature-taking under conditions different from this study.