A mesophilic, neutrophilic and aerobic, ammonia-oxidizing archaeon, strain EN76T, was isolated from garden soil in Vienna (Austria). Cells were irregular cocci with a diameter of 0.6–0.9 µm and possessed archaella and archaeal pili as cell appendages. Electron microscopy also indicated clearly discernible areas of high and low electron density, as well as tubule-like structures. Strain EN76Thad an S-layer with p3 symmetry, so far only reported for members of theSulfolobales. Crenarchaeol was the major core lipid. The organism gained energy by oxidizing ammonia to nitrite aerobically, thereby fixing CO2, but growth depended on the addition of small amounts of organic acids. The optimal growth temperature was 42 °C and the optimal pH was 7.5, with ammonium and pyruvate concentrations of 2.6 and 1 mM, respectively. The genome of strain EN76Thad a DNA G+C content of 52.7 mol%. Phylogenetic analyses of 16S rRNA genes showed that strain EN76Tis affiliated with the recently proposed phylumThaumarchaeota, sharing 85 % 16S rRNA gene sequence identity with the closest cultivated relative ‘CandidatusNitrosopumilus maritimus’ SCM1, a marine ammonia-oxidizing archaeon, and a maximum of 81 % 16S rRNA gene sequence identity with members of the phylaCrenarchaeotaandEuryarchaeotaand any of the other recently proposed phyla (e.g. ‘Korarchaeota’ and ‘Aigarchaeota’). We propose the nameNitrososphaera viennensisgen. nov., sp. nov. to accommodate strain EN76T. The type strain ofNitrososphaera viennensisis strain EN76T( = DSM 26422T = JMC 19564T).Additionally, we propose the familyNitrososphaeraceaefam. nov., the orderNitrososphaeralesord. nov. and the classNitrososphaeriaclassis nov.