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The environmental association of Cryptococcus neoformans var. gattii with decaying wood in tropical and subtropical regions of the world is well documented. In Australia, the yeast appears confined to certain species of Eucalyptus or very closely related tree species. In this study, we attempted to isolate C. n. var. gattii from different gum tree species in the Blue Mountains National Park. Out of 99 samples from 9 different tree species, only 3 yielded viable yeast colonies; 2 were from turpentine gums (Syncarpia glomulifera) and 1 was from a decayed stump of an unknown species. All of the colonized trees occurred in close proximity in urbanized areas of the Park, and all isolates shared identical DNA fingerprinting profiles. We suggest that domestic animal vectors may be responsible for the introduction and transmission of the yeast in this region, but that propagation and dispersal are very limited. This study indicates that C. n. var. gattii may occur on trees and in areas that were not previously expected to host it. However, the low incidence in the Blue Mountains National Park means this yeast is unlikely to pose any hazards to humans and animals living in or visiting this area.