• Applicability of an ATP assay for estimation of bioburden in caves.
  • A high variability of microbial biomass within a single cave system.
  • Cave infrastructures containing heavy metals reduce the viability of microbiota.
  • Underground habitats preserve and concentrate microbial biomass.


A commercially available adenosine triphosphate (ATP) detection system (Hygiena, USA), supported by cultivable microbial indicators, was used to estimate bioburden in different habitats in and outside show caves: air, water and solid surfaces. A strong positive correlation between ATP concentration expressed as Relative Light Units (RLU) and Colony-Forming-Units (CFU) was observed for swab samples from cave surfaces. In terms of ATP units, surfaces in a single cave system (Postojna Cave) varied considerably (240-1,258,800 RLU/ 20 cm2) and commonly exceeded the bioburden level of analogues on the surface (0-114,390 RLU/ 20 cm2). Cave sub-habitats were colonized by physiologically distinct microbial communities in terms of their nutrient demands, temperature requirements and r/K growth strategy. The highest ATP biomass indicator (1,258,800 RLU/ 20 cm2) for the speleothem that had been touched but accompanied with comparable concentration of CFU (~106 CFU/ 20 cm2) for other cave sub-habitats, can be related to the presence of deposited human epithelium skin cells. Show cave infrastructures containing heavy metals, e.g. copper used in safety fences, reduce the viability of microbiota. Mass cave visitation and the presence of allochthonous organic matter result in high levels of airborne and total biomass. Once such material becomes airborne, the location of its settling depends upon natural and human-induced air movements. Underground habitats play an important role in the preservation and concentration of microbial biomass using air and water as transport mechanisms.



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