The main objective of the present study was to demonstrate the effect of cumin and molasses supplementation on thermographic profile of lactating buffaloes at different anatomical locations viz., forehead temperature (°C), eye temperature (°C), ear temperature (°C), elbow joint temperature (°C) and flank temperature (°C) during hot dry and hot humid seasons season. A total of twenty four (24) lactating Murrah buffaloes, maintained at Kathura village of Sonipat district (India) were selected and monitored for the thermal profiling at different anatomical locations of body using infrared thermography at monthly interval during hot dry and hot humid seasons using an IR camera (ThermaCamTM SC2000;FLIR Systems, Inc., Wilsonville, OR, USA). Thermograms were analysed by ThermaCamTM Researcher 2001 software (FLIR Systems AB, Danderyd, Sweden). Statistical analysis revealed that different supplement and seasons had significant (P<0.05) effect on body surface temperature at different anatomical sites. The magnitude of the increase in the IRT temperature at different anatomical regions of lactating buffaloes was lower in the treatment groups compared to control group. The temperature of the eye region and the rectal temperature of lactating Murrah buffaloes were found to be closely related. The results of the study suggest that cumin and molasses can be used to alleviate heat stress in Murrah buffaloes. Further, infrared thermography of eyes can be used as non-invasive method for assessing the heat stress levels of lactating Murrah buffaloes without disturbing the animal welfare.