To define the neutral environmental temperature and assess the effects of deviation from that temperature on insensible water loss and heat balance, 12 premature infants were studied in a conventional incubator at four different predetermined ambient temperatures. Our method combines insensible water loss measured by a continuous read-out electronic scale with heat production as determined by open circuit measurement of oxygen consumption. An increase of 1 to 2 degrees C, to an ambient temperature above or near the top of the neutral zone, produced a significant rise in insensible water loss, from 1.90 +/- 0.76 to 3.08 +/- 1.19 ml/kg/hour (mean +/- SD), a corresponding rise in evaporative heat loss, and a fall in nonevaporative heat loss. A decrease of 1 to 2 degrees C, to a slightly subneutral ambient temperature, resulted in an increase in oxygen consumption from 5.82 +/- 0.92 to 7.45 +/- 1.50 ml/kg/minute, and an increase in total heat loss, but no change in insensible water loss and evaporative heat loss. The increased total heat loss was judged to be due entirely to a greater nonevaporative heat loss, both by convection and by radiation. The data confirm that ambient temperature is an important determinant of the magnitude and the partition of heat loss in low-birth-weight infants.