Use of biocides for the control of fungal outbreaks in subterranean environments: the case of the Lascaux Cave in France.

The Lascaux Cave in France suffered an outbreak of the fungus Fusarium solani in 2001. Biocides were applied for three years to control this outbreak. Four months after the initial biocide application, a new outbreak appeared in the form of black stains that progressively invaded the cave. The black stains on the ceiling and passage banks were so evident by 2007 that they became one of the cave's major problems. Therefore, biocides were used again in 2008. The present study investigated the fungal communities associated with the black stains and the effectiveness of the biocides applied, by using cloning, denaturing gradient gel electrophoresis, and culture-dependent methods. A novel species, Ochroconis lascauxensis, was the most abundant fungus in samples collected between 2007 and 2008, and the biocides applied were not effective in eliminating this fungus; on the contrary, they appeared to increase the fungal diversity. The fungal communities represented in the samples collected in 2010 were quite different from those collected in 2008 and 2009: the major OTUs corresponded to black yeasts belonging to the Herpotrichiellaceae family. The origin and evolution of these microorganisms are probably linked to the intensive biocide treatments and to the anthropogenic changes introduced by cave management.