A joint effort between the U.S. Geological Survey's (USGS) Global Desert Dust and NASA's Stratospheric and Cosmic Dust Programs identified culturable microbes from an air sample collected at an altitude of 20,000 m. A total of 4 fungal (Penicillium sp.) and 71 bacteria colonyforming units (70 colonies of Bacillus luciferensis believed to have originated from a single cell collected at altitude and one colony of Bacillus sphaericus) were enumerated, isolated and identified using a morphological key and 16S rDNA sequencing respectively. All of the isolates identified were sporeforming pigmented fungi or bacteria of terrestrial origin and demonstrate that the presence of viable microorganisms in Earth's upper atmosphere may not be uncommon.