It has been suggested that the discontinuous ventilation cycle (DVC) observed in many insects, including all ants described to date, is an adaptation to reduce respiratory water loss. To test this hypothesis, it is necessary to measure respiratory water loss as a percentage of total water loss and to estimate what sustained rates of water loss would be in the absence of spiracular control. We used two independent techniques to measure real-time water loss rates in female alates of Pogonomyrmex rugosus. The first measured water vapor emission and CO2 production simultaneously using dual- wavelength infrared absorbance analysis (DWIRAA). The second measured water loss gravimetrically. Real-time measurement allowed the separation of cuticular water loss rates (interburst) from water loss rates during the ventilation phase (burst) of the DVC. Cuticular permeability of P. rugosus female alates was only 27 ng h-1 cm-2 Pa-1, one-third of that reported for workers of the same species and the lowest yet reported for ants. Partly because of this low cuticular permeability, respiratory water loss represented a greater percentage of overall water loss (13 %) than has generally been reported for other insects. The DWIRAA and gravimetric techniques gave equivalent results. Peak rates of water loss during the burst phase were 2.8-fold higher than cuticular water loss rates alone (7.68 mg g-1 h-1 versus 2.77 mg g-1 h-1 at 25°C). This is a conservative estimate of water loss rates in the absence of spiracular control. Contrary to findings in certain other insects that suggest a negligible role for respiratory water loss, we find that, in an insect that employs the DVC and has low cuticular permeability, overall water loss rates rise several-fold in the absence of direct spiracular control. Our findings lend strong support to the water conservation hypothesis for the role of the DVC. In at least some insects, respiratory water loss rates can reach magnitudes significant enough, relative to other routes of water loss, for strong selective pressure to act on them.

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