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(1) Background: Airborne microbes are an integral part of a cave ecosystem. Cave allochtonous airborne microbiota, which occurs mainly during aerosolization from an underground river, from animals, and from visitors, is particularly pronounced in show caves. The impacts of tourists and natural river aerosolization on the cave air were estimated in large cave spaces within the Škocjan Caves; (2) Methods: Simultaneously with the measurements of atmospheric parameters, cultivable airborne bacteria were impacted, counted and identified using MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry); (3) Results: A mix of bacteria typically associated with humans and with natural habitats, including a large percentage of non-identified isolates, was found in the cave air. Few of the isolates were attributed to Risk Group 2. A strong positive correlation between tourist numbers and the rise in the concentration of airborne bacteria was indicated. Concentration of airborne bacteria rises to particularly high levels close to the underground river during periods of high discharge. A 10-times lower discharge reflected an approximately 20-times lower concentration of airborne bacteria; (4) Conclusions: Caves that are open and visited contain a diverse airborne microbiota originating from different sources. Enormous cave chambers that display relatively dynamic cave climate conditions do not normally support the enhancement of airborne bacterial concentrations

Spacio-Temporal Distribution and Tourist Impact on Airborne Bacteria in a Cave (Škocjan Caves, Slovenia)