Characterization in the archaeological excavation site of heterotrophic bacteria and fungi of deteriorated wall painting of Herculaneum in Italy.

Microbiological characterization of frescos in four different locations (Collegio degli Augustali, Casa del Colonnato Tuscanico, Casa dello Scheletro and Casa del Gran Portale) of excavation sites of Herculaneum was carried out. The use of infrared thermography allowed detecting sample points on frescos with greatest moisture not visible to the naked eye, resulting in structural damage. The microclimatic conditions provided perfect habitat for bacteria and fungi, particularly of spore forming and mould. In fact, heterotrophic bacteria were prevalent in all wall paintings monitored (ranging from 18 +/- 2 CFU 100 cm(-2) to 68 +/- 4 CFU 100 cm(-2)), whereas fungi were also detected but at lower levels (ranging from 9 +/- 2 CFU 100 cm(-2) to 45 +/- 3 CFU 100 cm(-2)). Cultural-based method allow us to identify by 16S and 26S rRNA partial sequence analysis heterotrophic microorganisms belonging to different genera of Bacillus and Aspergillus, Penicillium and Fusarium together with the unusual genera as Microascus and Coprinus. By using this approach, Bacillus-related species (B. cereus/B. thuringiensis group, B. simplex/B. muralis group, B. megaterium and B. subtilis) were isolated in all sample points analysed with the exception of the Casa dello Scheletro in which Micrococcus luteus/Arthrobactersp. group and Streptomyces fragilis were found. DGGE analysis of PCR amplified V3 region of rDNA from DNA directly recovered from frescos samples, enabled identification of bacterial species not identified using culturable technology asthose closest related to Microbacterium group, often associated with Brevibacterium, Streptomyces and Stenotrophomonas. Combination of culture-dependent and independent methods provided better microbiology characterization of heterotrophic microbiota present on the surface of ancient frescos of this important archaeological site.