Integrative biocodicology: a novel approach combining DNA analysis with FTIR and MicroHot table analysis for the characterisation of modern and ancient parchments

Abstract

This study shows for the first time the potential of combining the nanopore sequencing technology (MinION, ONT) with Whole Genome Amplification (WGA) to identify, simultaneously, endogenous DNA (of animal origin) and environmental DNA (from microbiome) in parchment. This DNA-based approach was integrated with Micro Hot Table (MHT) and Fourier Transform Infrared (FTIR) analysis to assess the deterioration of the parchment.

The novel strategy was tested on parchments from different ages: three modern parchments of known animal origin, a 19th-century fragment, and nine 14th-15th-century parchment drawings from the Graphic Collection of the Academy of Fine Arts in Vienna. The study compared micro-invasive (small piece of material) and non-invasive (PVC eraser rubbings) sampling. WGA with Nanopore sequencing successfully identified animal species and microbiome composition from both sampling approaches for the first time. The microbiome was dominated by bacteria, primarily human-associated genera (e.g., Staphylococcus, Streptococcus), but also environmental taxa (Acinetobacter, Chryseobacterium, Kaistella, Clostridium, and Pseudomonas), all well-known for their degradative capabilities, which is consistent with collagen degradation detected by MHT and FTIR. However, it also included viruses (Gemykrogvirus) and protozoa (Babesia). The animal origin was correctly determined for all modern parchment samples used as positive control, and it could also be inferred for the ancient parchment samples using both sampling approaches. The 19th-century parchment was found to be of caprine origin (Capra hircus), while all the ancient drawings were of bovine origin (Bos taurus). This approach merges molecular analysis and degradation study for comprehensive archival research.