Effects of Speleotherapy on Aerobiota: A Case Study from the Sežana Hospital Cave, Slovenia

Abstract

Speleotherapy is one of the non-pharmacological methods for the treatment and rehabilitation of patients with chronic respiratory diseases, especially those with chronic obstructive pulmonary disease (COPD) and asthma. On the one hand, one of the alleged main advantages of speleotherapeutic caves is the low microbial load in the air and the absence of other aeroallergens, but on the other hand, due to the lack of comprehensive air monitoring, there is little information on the pristine and human-influenced aerobiota in such environments. The aim of this study was to assess the anthropogenic effects of speleotherapy on the air microbiota and to investigate its potential impact on human health in Sežana Hospital Cave (Slovenia). From May 2020 to January 2023, air samples were collected in the cave before and after speleotherapeutic activities using two different volumetric air sampling methods—impaction and impingement—to isolate airborne microbiota. Along with sampling, environmental data were measured (CO2, humidity, wind, and temperature) to explore the anthropogenic effects on the aerobiota. While the presence of patients increased microbial concentrations by at least 83.3%, other parameters exhibited a lower impact or were attributed to seasonal changes. The structure and dynamics of the airborne microbiota are similar to those in show caves, indicating anthropization of the cave. Locally, concentrations of culturable microorganisms above 1000 CFU/m3 were detected, which could have negative or unpredictable effects on the autochthonous microbiota and possibly on human health. A mixture of bacteria and fungi typically associated with human microbiota was found in the air and identified by MALDI-TOF MS with a 90.9% identification success rate. Micrococcus luteus, Kocuria rosea, Staphylococcus hominis, and Staphylococcus capitis were identified as reliable indicators of cave anthropization.