We examined spontaneous locomotory behavior and respiratory pattern in replicate outbred populations of Drosophila melanogaster selected for desiccation resistance or starvation resistance, as well as their control and ancestral populations. Use of these populations allows us to compare evolved behavioral changes in response to different stress selections. We also reasoned that previously observed changes in respiratory patterns following selection for increased desiccation resistance might be associated with or even caused by changes in locomotory behavior. We measured spontaneous locomotory behavior using video recordings and a computer-based tracking system while simultaneously measuring patterns of CO(2) release from single fruit flies. Statistically significant differences in behavior were observed to be correlated with selection regime. Reduced levels of spontaneous locomotory activity were observed in moist air in both desiccation- and starvation-selected populations compared with their controls. Interestingly, in dry air, only the desiccation-selected flies continue to show reduced spontaneous locomotory activity. No correlation was found between the level of locomotory activity of individual flies and the respiratory patterns of those flies, indicating that the reduced activity levels that have evolved in these flies did not directly cause the documented changes in their respiratory pattern.