(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.

Biofilms of different colours that colonize surfaces within karst caves represent a source of nutrients. They occur commonly and abundantly at sites with sediments, and close to seepages or underground rivers. Golden-yellow subaerial biofilms are particularly well observed because of their contrast with their surroundings, the characteristics of the pigment and recently, even more, due to the characteristics of light-emitting diode (LED) illumination. Yellow microbial biofilms were sampled from three caves in southwestern Slovenia, Dimnice, Križna jama and Sveta jama. The highest concentration of cultivable microbes (2.33×108 CFU/g) and the biggest number of identified bacteria (66.0%) were retrieved from a sample from Sveta jama. Using MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight) for bacterial identification showed that different species of Pseudomonas prevailed in all samples. Yellow biofilms showed an absorption peak around 400 nm, and two emission peaks, a major in the blue (~460 nm) and a minor in the orange (~600 nm) parts of the spectrum when excited at 405 nm. Microbial mats that colonize surfaces are probably frequently overlooked in caves because they are difficult to observe when they have no pigmentation and the contrast with their surroundings is low. Additional studies are needed to aid the understanding of the role of pigmented biofilms and their interactions with underlying substrata in respect of the evolution of substrate micromorphology.