Deception Island is an Antarctic Specially Managed Area that houses historically important sites such as the remains of historical wooden buildings. The impacts of fungal communities on wood in polar historical sites have been investigated, but little is known of the impacts of other eukaryote groups. In the current study we used high-throughput sequencing to investigate the diversity of non-fungal eukaryotic organisms present in wood samples from Whalers Bay. Four sites were sampled, and DNA sequences representing three kingdoms (Chromista, Protozoa and Viridiplantae) and four phyla (Ciliophora, Perclozoa, Chlorophyta and Magnoliophyta) were identified, representing a total of 43 taxa. Biscoe House Annex hosted the richest diversity, with 20 taxa, followed by the whaling boat, Biscoe House and the Hunting Lodge, with 16, 15 and 12 taxa, respectively. The most frequently detected sequences were assigned to the ciliate group Sporadotrichida, some of which are known to play a role in cellulose degradation. Among the Chlorophyta, the sequences detected included common taxa previously recorded, but the flowering plant data represented only exotic taxa, probably associated with human activity or airborne transfer. The use of high-throughput sequencing provided valuable data on communities associated with anthropogenically sourced and now decaying wood in Antarctica.