ABSTRACT. Objective. To estimate fat and energy contents of human milk during prolonged lactation.Methods. Thirty-four mothers, of term, healthy, growing children, who had been lactating for >1 year (12-39 months) were recruited. Control subjects were 27 mothers, of term infants, who had been lactating for 2 to 6 months. Fat contents of the milk samples were estimated as creamatocrit (CMT) levels. Energy contents of the milk were measured with a bomb calorimeter.Results. The groups did not differ in terms of maternal height and diet, infant birth weight, gestational age, or breastfeeding frequency. They differed significantly in terms of maternal age, maternal weight, and BMI. The mean CMT levels were 7.36 ؎ 2.65% in the short-duration group and 10.65 ؎ 5.07% in the long-lactation group. The mean energy contents were 3103.7 ؎ 863.2 kJ/L in the short-duration group and 3683.2 ؎ 1032.2 kJ/L in the long-duration group. The mean CMT levels and mean energy contents were correlated significantly with the duration of lactation (R 2 ؍ 0.22 and R 2 ؍ 0.23, respectively). In multivariate regression analysis, CMT levels (or energy contents) were not influenced by maternal age, diet, BMI, or number of daily feedings but remained significantly influenced by the duration of lactation.Conclusions. Human milk expressed by mothers who have been lactating for >1 year has significantly increased fat and energy contents, compared with milk expressed by women who have been lactating for shorter periods. During prolonged lactation, the fat energy contribution of breast milk to the infant diet might be significant. T he optimal duration of breastfeeding is unknown. The American Academy of Pediatrics recommends exclusive breastfeeding for 6 months and a total duration of Ն1 year to obtain the "full benefits of breastfeeding." 1 Among frequently recognized long-term benefits of breastfeeding are reductions in cardiovascular risks in adulthood. 1,2 These reductions were challenged by a retrospective epidemiologic study of men born in 1920 to 1930 in Hertfordshire, England, which suggested that the beneficial effects of breastfeeding on cardiovascular risks existed as long as weaning was performed before 1 year of age; after that time, continued breastfeeding was associated paradoxically with increased cardiovascular risks. 2 Moreover, a study by Leeson et al 3 suggested that prolonged breastfeeding might lead to unwelcome outcomes and might even increase cardiovascular risks in adulthood.In developed countries, a minority of women continue to lactate for Ͼ1 year; in one study from Italy, 17% of mothers were still breastfeeding at 12 months after delivery. 4 The energy contribution of human milk (HM) to the diet of partially breastfed children beyond the first year of life is unknown, because there are no data on the amounts of HM consumed by these children and the nutritional content of HM after prolonged breastfeeding is little known. In particular, the fat and energy contents of HM after prolonged breastfeeding have not been analyz...