Fasting heat production (FHP) is used to assess the maintenance net energy requirement of animals. Herein, the FHP of layer-type pullets was estimated. In trial 1, 16 40-day-old Jingfen layer-type pullets were divided into 4 groups of 4 chickens and placed in 4 respiratory chambers. Pullets had free access to feed and water. After 4-d acclimatization, feed was withdrawn, and chickens were measured for FHP for 3 consecutive days. In trial 2, twenty-four 40-day-old pullets were placed in 4 respiratory calorimetry chambers, with 6 pullets per chamber. After 4-d acclimatization, one chamber was randomly selected and all pullets in the chamber was sampled at 5, 25, 50, or 65 h after feed withdrawal. The result showed that FHP declined with fasting time and reached the lowest level between 48 and 72 h. Respiratory quotient was decreased (P < 0.05) between 24 and 48 h compared with that in the first 24 h after fasting. The FHP in the light period showed a significant to decline with fasting time (P < 0.01), whereas the FHP in the dark period was decreased (P < 0.01) 24 h after fasting. Body weight, thigh mass, and abdominal fat decreased (P < 0.05) at 25 h after fasting. Serum glucose were increased (P < 0.01) and while triglycerides were significantly decreased (P < 0.01) at 50 h compared with that at 5 and 25 h time point. The result suggests that the adequate measuring period for FHP for layer-type pullets is from 24 to 48 h after fasting. The FHP of 7-wk-old layer-type pullets was 562.20 kJ/kg of BW0.75/d under a 10-h light and 14-h dark lighting regime.