Cryoconite holes are small, extreme habitats, widespread in the ablation zones of glaciers worldwide. They can provide a suitable environment for microorganisms including bacteria, cyanobacteria, algae, fungi, and invertebrates. Diatoms have been previously recovered from cryoconite holes of Greenland and of Svalbard, and recent findings from Antarctica suggest that cryoconite holes may harbor a unique diatom flora distinct from other aquatic habitats nearby. In the present study, we characterize the diatom communities of Nordenskiöld glacier cryoconite holes in Billefjorden (Svalbard, Spitsbergen), and multivariate approaches were used to compare them with three freshwater localities in the immediate vicinity to investigate possible sources of the species pool. We found cryoconite holes to have similar or greater average genus-richness than adjacent lake/ ponds habitats, even though lower numbers of valves were recovered. Overall, cryoconite hole diatom communities differed significantly from those observed in lakes, suggesting that other sources actively contribute to these communities than nearby lakes alone. This further suggests that (i) diatoms present in cryoconite might not exclusively originate from aquatic habitats, but also from (semi-) terrestrial ones; and (ii) that a much wider area than the immediate surroundings should be considered as a possible source for cryoconite diatom flora