Riparian forests provide important roosting habitat, abundant prey and access to drinking water for many bat species but to date there has been little research on the differential quality of habitats within riparian areas. We quantified the density of potential roost cavities in three age classes (i.e., young: ca. 20 years, mature: ca. 60 years, and old: ca. 100 years) of riparian cottonwood (Populus deltoides) forest stands. Bat activity was also sampled using acoustic detectors in one representative stand of each age class. Stands were situated along an 80 km stretch of the Missouri River in southeastern South Dakota and northwestern Iowa, USA. We predicted the highest density of potential roosts and the highest activity of bats to occur in the oldest age class. Contrary to our predictions, and previous work in aspen dominated upland sites, we found that the density of potential roosts was not significantly different between mature and old stands. However, there were no potential roosts in young stands. Data from guano traps verified the use of a number of cavities in both mature and old stands. Both commuting and foraging activities were highest in the mature, relative to the old and young stand. In total, our data indicate that mature and old stands represent high quality roosting habitat, with the mature being used preferentially for commuting and foraging. Trees in the oldest stands, however, are nearing the end of their lifespan and falling. Younger cohorts must therefore be retained for future recruitment of natural cavities.