Fungal spore loads in the air of cool-temperature, temperate, and tropical aviaries were collected with an Andersen N-6 air sampler. The relationship of spore loads to air-handler and exhibit design in these three environments was examined. In addition, a 2-yr longitudinal study of fungal spore loads in the air of a newly designed and installed air-handling system in the R. J. Reynolds Forest Aviary at the North Carolina Zoological Park was compared to the earlier air-handling system that it replaced. High-efficiency particulate air filters installed in cool-temperature aviaries produced the cleanest air, although pleated filters showed only marginally higher spore loads. Temperate and tropical aviaries with pleated filters or bag filters with variable-velocity fans had much higher spore loads. Tropical and temperate exhibits with bag filters and constant-velocity fans produced the cleanest air in tropical and temperate exhibits. Information on the relative effectiveness of different air-handling system designs and related costs/benefits should be used by zoo managers when they are designing or retrofitting aviary air-handling systems.