Seven pairs of young adult male identical twins completed a negative energy balance protocol during which they exercised on cycle ergometers twice a day, 9 out of 10 days, over a period of 93 days while being kept on a constant daily energy and nutrient intake. The total energy deficit caused by exercise above tlie estimated energy cost of body weight maintenance readied 244 f 9.8 MJ (Mean f SEM).Baseline energy intake was estimated over a period of 17 days preceding the negative energy balance protocol. M e a n body weight loss was 5.0 kg (SEM = 0.6) (p < 0.001) and it was entirely accounted for by tlie loss of fat mass (p < 0.001). Fat-free mass was unchanged. Body energy losses reached 191 MJ (SEM = 24) (p < 0.001) wliicli represented about 78% of the estimated energy deficit. Subcutaneous fat loss was sliglitly more pronounced on the trunk than on the limbs as estimated from skinfolds, circumferences, and computed tomograpliy (CT). The reduction in CT-assessed abdominal visceral fat was quite striking, from 81 cm2 (SEM = 5) to 52 cm2 (SEM = 6) (p < 0.001). A t tlie same submaximal power output level, subjects oxidized more lipids than carbohydrates after tlie program as indicated by tlie changes in the respiratory exchange ratio (p 10.05). Intrapair resemblance was observed for tlie changes in body weight (p < 0.05), fat mass (P < 0.01), percent fat (p < 0.01), body energy content (p < 0.01), sum of 10 skinfolds (p < 0.01), abdominal visceral fat (p < 0.01), fasting plasma triglycerides (p < 0.05) and cliolesterol (p < 0.05), maximal oxygen uptake (p < 0.05), and respiratory exchange ratio during submaximal work (p < 0.01). We conclude that even though there were large individual differ- ences in response to the negative energy balance and exercise protocol, subjects with tlie same genotype were more alike in responses than sulljects with different genotypes particularly for, body fat, body energy, and abdominal visceral fat changes. High lipid oxidizers and low lipid oxidizers during submaximal exercise were also seen despite the fact that all subjects liad experienced tlie same exercise and nutritional conditions for about three montlis.