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In this study, a culture-independent Illumina MiSeq sequencing strategy was applied to investigate the microbial communities colonizing the ancient painted sculptures of the Maijishan Grottoes, a famous World Cultural Heritage site listed by UNESCO in China. Four mixed samples were collected from Cave 4-4 of the Maijishan Grottoes, the so-called Upper Seven Buddha Pavilion, which was built during the Northern Zhou Dynasty (557-581AD). The 16/18S rRNA gene-based sequences revealed a rich bacterial diversity and a relatively low fungal abundance, including the bacterial groups Actinobacteria, Acidobacteria, Bacteroidetes, Cyanobacteria, Chloroflexi, Firmicutes, Proteobacteria and Verrucomicrobia and the fungal groups Ascomycota, Basidiomycota and Chytridiomycota. Among them, the bacteria genera of Pseudonocardia and Rubrobacter and unclassified fungi in the order of Capnodiales were dominant. The relative abundance of Pseudonocardia in the painted layer samples was higher than that in the dust sample, while Cyanobacteria dominated in the dust sample. Many of them have been discovered at other cultural heritage sites and associated with the biodeterioration of cultural relics. The presence and activity of these pioneering microorganisms may lead to an unexpected deterioration of the painted sculptures that are preserved in this heritage site. Thus, proper management strategies and potential risk monitoring should be used in the Maijishan Grottoes to improve the conservation of these precious painted sculptures.

The Dadiwan F901 site, boasting a history of over 5000 years, stands as the largest and most intricately crafted large-scale housing structure from China’s prehistoric era. The early renovation efforts, incorporating a sealed glass curtain wall, led to a continuous rise in relative humidity within the site, triggering outbreaks of microbial diseases. Subsequent measures successfully restored stability to the thermal and humid environment. This paper employs on-site real-time environmental monitoring and numerical simulation methods to assess the ventilation effectiveness and relative humidity changes before and after multiple interior modifications of the Dadiwan F901 site museum. The results indicate that the fully enclosed glass curtain wall can suppress the dependence of indoor humidity fluctuations on external weather fluctuations but has generated unintended consequences, leading to increased air relative humidity and even reaching saturation in the museum space. The strategic deployment of louvered windows and duct fans proved effective in enhancing internal airflow dynamics and overall air exchange capacity. It was possible to ensure that the relative humidity inside the site remained at approximately 70%, meeting the essential requirements for the preservation of cultural relics. This study is of great significance for alleviating the deterioration problem of enclosed exhibition halls of earthen relics.

Microbial colonization on stone monuments leads to subsequent biodeterioration; determining the microbe diversity, compositions, and metabolic capacities is essential for understanding biodeterioration mechanisms and undertaking heritage management. Here, samples of epilithic biofilm and naturally weathered and exfoliated sandstone particles from different locations at the Beishiku Temple were collected to investigate bacterial and fungal community diversity and structure using a culture-based method. The biodeterioration potential of isolated fungal strains was analyzed in terms of pigmentation, calcite dissolution, organic acids, biomineralization ability, and biocide susceptibility. The results showed that the diversities and communities of bacteria and fungi differed for the different sample types from different locations. The population of culturable microorganisms in biofilm samples was more abundant than that present in the samples exposed to natural weathering. The environmental temperature, relative humidity, and pH were closely related to the variation in and distribution of microbial communities. Fungal biodeterioration tests showed that isolated strains four and five were pigment producers and capable of dissolving carbonates, respectively. Their biomineralization through the precipitation of calcium oxalate and calcite carbonate could be potentially applied as a biotechnology for stone heritage consolidation and the mitigation of weathering for monuments. This study adds to our understanding of culturable microbial communities and the bioprotection potential of fungal biomineralization.

[This corrects the article DOI: 10.1371/journal.pone.0179718.].

Microbial deterioration as one of the widespread problems in archaeological site museums significantly affects their safety and exhibits. This paper systemically investigated the environments and conditions of microbial outbreaks in the Dadiwan No. F901 site museum, which is a representative archaeological site of prehistoric Yangshao culture. The morphology and harmful characteristics of the outbreak microorganisms were analyzed by microscopic techniques. The ultraviolet resistance of harmful microorganisms was also studied. Combining these findings with the original facilities of the site museum, a scientific and reasonable project was proposed to control and prevent the activity of harmful microorganisms. In addition, a 1% OIT/DCOIT biocide concentration was applied to inhibit microorganism-caused deterioration, in combination with mechanical removal based on laboratory tests and screening in situ. The effectiveness of microbial control was assessed using a portable microscope, ATP bioluminescence assay, and color difference detection. As a long-lasting preventive measure for microbial deterioration, an ultraviolet sterilization system can efficaciously prevent the re-outbreak of microbial deterioration to form a relatively stable dynamic balance for the surroundings of the site. This study is a resultful exploration in terms of microbial control and plays an important role in the sustainable protection of archaeological site museums.

White and black biofilms threatened the wall paintings in UNESCO-listed Maijishan Grottoes. This study investigates these biofilms for viability, composition, functional groups, and degradation potential. The results revealed Scopulariopsis and unclassified_Eurotiomycetes as the core fungal taxa in black and white biofilms, respectively. Fluorescence microscopy and RNA-based high-throughput sequencing proved that white biofilms exhibited higher viability. In addition, we isolated Arachnomyces sp. (unclassified_Eurotiomycetes) as the dominant strain in white biofilms. Laboratory assays demonstrated that Arachnomyces sp. has strong capacities to degrade cellulose, gelatin, casein, and keratin, indicating its biodeterioration of wall paintings. Furthermore, black biofilms were dominated by dung saprotrophic fungi, likely introduced through animal activities within the grottoes. Our findings underscore that the divergences in fungal composition between black and white biofilms are driven by microenvironmental factors, substrate properties, animal activities and ecological selections. The interplay of microbial dispersal and environmental selection shapes biofilms, and microenvironmental control is the essential targeted strategy.