Development of protocols for microbiological control in Altamira cave

Abstract

Art Caves are threatened by uncontrolled growth of microorganisms, especially fungi. At the same time, in spite of the extreme ambiental conditions, they are inhabited by very diverse microorganisms. Altamira Cave contains one of the best collections of parietal art and is also threaten by potential microbial deterioration. In order to preserve the state of paintings, after a research project developed in 2013-2014, came out the “Plan de Conservación Preventiva” (PCP) for the Altamira Cave. Among the objectives of the PCP is the systematic control of the cave microbiota, and methods investigated here will become standard for these purposes. Cultivation of bacteria from natural habitats shows that only a small fraction of the bacteria present in such environments, can be cultivated under regular laboratory conditions and clearly indicate that alternative methods are required for quantitative purposes. Therefore, we have used alternative culture methods, as well as other quantitative methods based on fluorescent staining of microbes such as direct counting by fluorescence microscopy and flow cytometry. These techniques have been applied to water, air and soil samples and the results compared and combined with qualitative biodiversity analysis performed by 16S rDNA sequencing from isolated colonies or directly from the cave samples.