By simultaneously measuring carbon dioxide release, water loss and flight force in several species of fruit flies in the genus Drosophila, we have investigated respiration and respiratory transpiration during elevated locomotor activity. We presented tethered flying flies with moving visual stimuli in a virtual flight arena, which induced them to vary both flight force and energetic output. In response to the visual motion, the flies altered their energetic output as measured by changes in carbon dioxide release and concomitant changes in respiratory water loss. We examined the effect of absolute body size on respiration and transpiration by studying four different-sized species of fruit flies. In resting flies, body-mass-specific CO(2) release and water loss tend to decrease more rapidly with size than predicted according to simple allometric relationships. During flight, the mass-specific metabolic rate decreases with increasing body size with an allometric exponent of -0.22, which is slightly lower than the scaling exponents found in other flying insects. In contrast, the mass-specific rate of water loss appears to be proportionately greater in small animals than can be explained by a simple allometric model for spiracular transpiration. Because fractional water content does not change significantly with increasing body size, the smallest species face not only larger mass-specific energetic expenditures during flight but also a higher risk of desiccation than their larger relatives. Fruit flies lower their desiccation risk by replenishing up to 75 % of the lost bulk water by metabolic water production, which significantly lowers the risk of desiccation for animals flying under xeric environmental conditions