Aspergillosis is a common cause of mortality in captive birds, particularly in recently imported birds or captive chicks and their parents. Use of the Andersen N-6 single-stage viable air sampler in the North Carolina Zoological Park (NCZP) R.J.R. Nabisco Rocky Coast Alcid Exhibit before and after the introduction of birds allowed a unique study of the mycological content of the air in a developing self-contained ecosystem. The Alcid Exhibit had a median count of 17 colony-forming-units (CFU)/m3 of air in comparison to 200-500 CFU/m3 and 1,000-3,500 CFU/m3 reported in human dwellings and the NCZP R.J. Reynolds Forest Aviary, respectively. Cladosporium and Penicillium represented 21.3% and Aspergillus 1.08% of the fungi collected. During the study, no respiratory mycoses were reported in any of the alcids. Continuous high-efficiency particulate air filtration, maintenance of low exhibit air temperatures, and an environment with little residual organic material capable of supporting fungal growth were important factors contributing to low colony counts. All colony counts >100 CFU/m3 in the exhibit were related to the apparent introduction of fungi from outside the facility. A reduction in the number of fungi transported from an external source into enclosed cool-temperature aviaries may be sufficient to avoid outbreaks of aspergillosis.