Aspergillus fumigatus is a saprophytic, filamentous fungus that is ubiquitous outdoors (soil, decaying vegetation) and indoors (hospitals, simulated closed habitats, etc.). A. fumigatus can adapt to various environmental conditions and form airborne conidia that are the inoculum for a variety of diseases (e.g. non- and invasive pulmonary infections, allergic bronchopulmonary aspergillosis, etc.) in immunocompromised hosts. In an on-going Microbial Observatory Experiments on the International Space Station (ISS) molecular phylogeny of several fungal isolates were characterized. Two strains, ISSF 21 and IF1SW-F4, were isolated from the HEPA filter and the surface of the Cupola of the ISS, respectively. Using primers targeting the internal transcribed spacers ITS1 and 2, both isolates were identified as A. fumigatus. The whole genome sequence analysis of ISSF 21 revealed increased number of single nucleotide polymorphisms (SNPs) when compared to the reference A. fumigatus 293. Knowing that A. fumigatus is an opportunistic pathogen and microgravity highly influences the antibiotic susceptibility and pathogenicity of microorganisms, we examined pathogenicity of both ISS isolates using the zebrafish larval model. The space station isolates (ISSF-021 and IF1SW-F4) were more virulent than two clinical strains (Af293 and CEA10) whose pathogenicity was highly characterized. Here the whole genome sequences of ISSF-021 strain are being deposited.