Utilization of three milk diets including cream, casein or whey, each naturally labelled with 13C (1 mmol 13C excess) from C4 sources, by six young male calves of the Deutsche Fleckvieh breed was investigated in a Latin-square split-plot design. Each milk diet was examined under resting conditions and during a short period of physical exercise on a treadmill. δ13C values (‰) in carbon dioxide in expired air were measured at intervals of about 1 h during 6·5 h after food intake. Expired air samples for CO2 isolation, subsequent isotopic analysis, measurement of CO2 production and respiratory quotient were taken at about hourly intervals and 13C recovery rates over 6·5 h were calculated. Feeding milk containing enriched milk casein, cream, or whey resulted in maximal significant 13C enrichments over background (δ13C) in CO2 of +1, +2·4 and +2·2‰, and recovery rates of 3·6, 9·9 and 12·2% respectively. This comparison shows the different kinetic behaviour of the main nutrients during the oxidation in tissues. The short exercise period (5 min at 1 J/s per kg body-weight + 5 min at 2 J/s per kg body-weight) did not influence the recovery rates significantly. However, after 10 min of muscular exercise there was a brief decrease in δ13C value of expired air which disappeared within the first 5 min of rest. These experiments demonstrate for the first time the applicability of the 13C breath test with naturally enriched diets in animal nutrition research and that quantitative results may be obtained.