Waterlogged archaeological wood is exposed to a high risk of biological degradation during the post-excavation phases of storage and restoration. For this reason, often biocides must be used to preserve wooden remains. In the present work three essential oils (cinnamon, wild thyme, and common thyme) were tested as possible alternative biocides to use in the preservation of waterlogged archaeological wood. The oils were first tested in vitro to establish the minimum inhibitory concentration (MIC) and to evaluate the biocidal activity on selected fungal strains. Then, the established MIC was applied on waterlogged archaeological wood samples and during an actual restoration treatment. The effectiveness of the oils was evaluated through cultural analyses, ATP quantification, and next-generation sequencing. The results showed that the oils caused a significant decrease in the vitality of fungal mycelia grown in vitro and of the microbiota present in treated wood and storage water. Furthermore, an influence on the composition of the bacterial communities of treated wood samples was observed. Although further tests are needed to evaluate interferences with the materials used during restoration procedures, essential oils could be considered as a possible alternative to the currently used biocide.

Waterlogged archaeological wood (WAW) is considered a precious material, first-hand account of past civilizations. Like any organic material, it is subjected to biodegradative action of microorganisms whose activity could be particularly fast and dangerous during the phases of excavation, storage and restoration. The present work aimed to characterize the microorganisms present in WAW during these tricky periods to evaluate the biological risk it is exposed to. The bacterial and fungal communities inhabiting woods coming from two archaeological sites (Pisa and Naples) were investigated through Next-Generation Sequencing (NGS). High-throughput sequencing of extracted DNA fragments was performed using the reversible terminator-based sequencing chemistry with the Illumina MiSeq platform. The analyses revealed that the two archaeological sites showed distinct richness and biodiversity, as expected. In all the WAWs, the bacterial community harbored mainly Proteobacteria, whereas Bacteroidetes was well represented only in Naples communities and taxa belonging to the phyla Chloroflexi only in the Pisa site. Concerning the fungal community, the two sites were dominated by different phyla: Ascomycota for Naples samples and Basidiomycota for Pisa. Interestingly, most of the identified bacterial and fungal taxa have cellulolytic or ligninolytic ability. These results provide new and useful background information concerning the composition of WAW microbiota and the threat it represents for this precious material.