Heterotrophic microorganisms in deteriorated medieval wall paintings in southern Italian churches

The Campania region in southern Italy is noted for its large number of churches that harbour invaluable frescoes, dated from the beginnings of the 4th up to the 13th century. The wall paintings represent an integral part of the monuments, and their deterioration constitutes a potentially significant loss for the world's cultural heritage. Heterotrophic microorganisms such as bacteria and mould can grow on the surface of paintings that contain a wide range of organic and inorganic constituents, and provide different ecological niches that are exploited by a large variety of microbial species. We isolated and identified the heterotrophic microorganisms found in the biodegraded medieval wall paintings of seven historical churches in Campania. The paintings showed different levels of microbial contamination. Microbiological analysis of different paintings gave an overview of the different heterotrophic microorganisms. Bacteria and moulds were isolated from 77% of the sampling points analysed, in which the most common type of alteration was discolouration often associated with detachment of the paint layer. Bacterial strains were identified by 16S rRNA partial sequence analysis. The Bacillus genus was isolated in all churches, even though the type of species was variable, whereas all actinomycetes strains, isolated in five of the seven churches analysed, could be referred to the Streptomyces genus. The similarity of the sequences analysed of the 42 Bacillus spp., 2 Paenibacillus spp. and reference strains of different species showed that these bacteria differentiated in 14 groups. The most frequently occurring taxa were most closely related to Bacillus cereus/thurigiensis/anthracis and Bacillus pumilus groups. Thirteen Streptomyces spp. were differentiated in seven groups on the basis of neighbor-joining analysis of 16S rRNA. Fungi belonging to the genera Penicillium, Aspergillus, Fusarium and Alternaria were also isolated from deteriorated wall paintings.