Biodeterioration is a serious threat to cultural heritage objects and buildings. The deterioration of a given material often incurs irreparable losses in terms of uniqueness and historical value. Hence preventive actions should be taken. One important challenge is to identify microbes involved in the biodeterioration process. In this study, we analyzed the microbial diversity of an ancient architectonical structure of the Rotunda of Sts. Felix and Adauctus, which is a part of the Wawel Royal Castle located in Krakow, Poland. The Rotunda is unavailable to tourists and could be treated as an extreme habitat due to the low content of nutrients coming either from sandstone plates bound with lime mortar or air movement. Microbial diversity was analyzed with the use of the high-throughput sequencing of marker genes corresponding to fragments of 16S rDNA (for Bacteria) and ITS2 (internal transcribed spacer 2) (for Fungi). The results showed that the microbial community adhered to wall surfaces is, to a large extent, endemic. Furthermore, alongside many microorganisms that could be destructive to masonry and mortar (e.g., Pseudomonas, Aspergillus), there were also bacteria, such as species of genera Bacillus, Paenisporosarcina, and Amycolatopsis, that can positively affect wall surface properties by reducing the damage caused by the presence of other microorganisms. We also showed that airborne microorganisms probably have little impact on the biodeterioration process as their abundance in the microbial community adhered to the ancient walls was very low.
Microorganisms are known to colonize and modify various environments, including cultural heritage buildings. Anthropogenic indoor environments often provides specific conditions for microbial growth e.g. microclimatic parameters. These conditions are potentially easy to control thus it is advisable to characterize and monitor bacterial and fungal communities in indoor habitats. Microbiological pollution in museums is particularly important because bacteria and fungi contribute to the deterioration of cultural heritage objects and may have a negative impact on people’s health. In our study we performed microbiological analysis of Wawel Royal Castle in Krakow, which is one of the greatest Polish national heritage, and was visited by over 1.4 million tourists in 2017. There are large and valuable collections of objects related to the Polish rulers e.g. rare tapestries and paintings. Microbiome analysis was done by culture-dependent experiments for I) airborne microorganisms (50 litres of air) sampled onto various types of agar media, as well as culture-independent analysis of II) airborne microorganisms (12000 litres of air) sampled onto saline, and III) microbes from dust from historical surfaces. Microbiological diversity was determined by analysis of deep sequenced amplicons covering hypervariable regions V3V4 of 16S rDNA gene and ITS2 region located between 5.8S and 26S rDNA.
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