A time travel story: metagenomic analyses decipher the unknown geographical shift and the storage history of possibly smuggled antique marble statues

Abstract

In this study, three possibly smuggled marble statues of an unknown origin, two human torsi (a female and a male) and a small head, were subjected to molecular analyses. The aim was to reconstruct the history of the storage of each single statue, to infer the possible relationship among them, and to elucidate their geographical shift. A genetic strategy, comprising metagenomic analyses of the 16S ribosomal DNA (rDNA) of prokaryotes, 18S rDNA of eukaryotes, as well as internal transcribed spacer regions of fungi, was performed by using the Ion Torrent sequencing platform. Results suggest a possible common history of storage of the two human torsi; their eukaryotic microbiomes showed similarities comprising many soil-inhabiting organisms, which may indicate storage or burial in land of agricultural soil. For the male torso, it was possible to infer the geographical origin, due to the presence of DNA traces of Taiwania, a tree found only in Asia. The small head displayed differences concerning the eukaryotic community, compared with the other two samples, but showed intriguing similarities with the female torso concerning the bacterial community. Both displayed many halotolerant and halophilic bacteria, which may indicate a longer stay in arid and semi-arid surroundings as well as marine environments. The microbiomes retrieved from each statue showed to be very specific, but some individual members showed to be biological markers for the geographical regions through which the pieces traveled and for the conditions under which they were stored.

Decoding the biological information contained in two ancient Slavonic parchment codices: an added historical value

“…In this context, it is important to note that metagenomics refers to untargeted sequencing analyses, the so‐called ‘shotgun approach’, which consists of sequencing the entire DNA library representing all the molecular components of a given sample (Teasdale et al ., 2017). In contrast, a second procedure more commonly applied in microbial ecology studies through NGS analyses, focuses on specific conserved sequences such as ribosomal RNA genes (Piñar et al ., 2019). The latter approach called the ‘amplicon sequencing approach’ has several advantages, such as less complexity of the data obtained, a larger number of sequences that can be assigned to a specific organism or a group of related organisms and, in addition, it can be applied for ‘target enrichment’ both during library preparation and after it for selecting particular DNA fragments of interest, especially when working with ancient DNA (Vai et al ., 2017).…”

“…Nowadays, high-throughput molecular methods enable the extraction and the analysis of the smallest traces of DNA from ancient objects and by the help of bioinformatics and international databases, to assign single molecules of DNA to certain animals, plants, bacteria, fungi or even to humans. The compilation of these data results in an individual microbiome or 'biological pedigree' (Piñar et al, 2019(Piñar et al, , 2020, which can be considered as an historical added value. The recovered biological information offers new possibilities to answer relevant questions related to valuable objects made of parchment, such as the book making process, the skin/animal type used to produce the parchment, its conservation status and its historical use (Teasdale et al, 2015(Teasdale et al, , 2017Migliore et al, 2017Migliore et al, , 2019Fiddyment et al, 2019).…”

The Microbiome of Leonardo da Vinci’s Drawings: A Bio-Archive of Their History

“…However, any object already contains DNA, from the people who have handled the object since its creation and from all the organisms and microorganisms that have come into contact with it, also through the deposition of dust and dirt. This natural bioarchive changes with time and can give certain information on some particular events, such as the presence on the materials of deteriorating microorganisms harmful for conservation, but also the permanence of the object in particular geographical locations, as indicated by the molecular traces left by the pollen of endemic plants (Piñar et al, 2019).…”

“…Although molecular techniques are not completely free from bias (Sterflinger et al, 2018a), the establishment of nextand third-generation sequencing technologies in the field of cultural heritage is providing a powerful DNA-based approach to monitor the preservation status as well as to foresee the risk of deterioration of valuable objects (Piñar et al, 2020a). The data generated can provide interesting information that can contribute to surprising insights into the objects being investigated, such as the selection of materials at the time of manufacture and the conditions of their storage or about possible displacements and geographical origins, but also uncover important information about the object's history of use (Teasdale et al, 2017;Piñar et al, 2019Piñar et al, , 2020b. This information may help to understand many open questions in a variety of fields, as archeology, history, restoration, philology, criminology but also, they can contribute to giving an historical benefit to the investigated objects in form of a bio-archive (Piñar et al, 2019(Piñar et al, , 2020b.…”

“…The data generated can provide interesting information that can contribute to surprising insights into the objects being investigated, such as the selection of materials at the time of manufacture and the conditions of their storage or about possible displacements and geographical origins, but also uncover important information about the object's history of use (Teasdale et al, 2017;Piñar et al, 2019Piñar et al, , 2020b. This information may help to understand many open questions in a variety of fields, as archeology, history, restoration, philology, criminology but also, they can contribute to giving an historical benefit to the investigated objects in form of a bio-archive (Piñar et al, 2019(Piñar et al, , 2020b.…”