Changes in milk composition associated with maternal dietary obesity and cafeteria feeding were investigated. Protein, lactose and fat contents, and the fat composition, were determined for lean and obese rats given a cafeteria diet at different stages of reproduction. Feeding the cafeteria diet during lactation resulted in an increase in long-chain fatty acids and a fall in the characteristic medium-chain fatty acids. This effect was modified by obesity and the diet during pregnancy. Feeding the cafeteria diet in lactation reduced the milk protein and increased the fat. The milk of obese rats contained more energy, with more fat but less protein than that of lean rats. Increases in fat and long-chain fatty acid content, and decreases in protein and medium-chain fatty acid content of the milk were correlated with increased maternal intake of energy, total fat and long-chain fatty acids. Thus, the greatest influence on milk composition is exerted by the maternal diet during lactation. However, these effects are modified by pre-existing maternal obesity and the diet during pregnancy.