The influence of human exploration on the microbial community structure and ammonia oxidizing potential of the Su Bentu limestone cave in Sardinia, Italy

The bacterial diversity in the Su Bentu Cave in Sardinia was investigated by means of 16S rRNA gene-based analysis. This 15 km long cave, carved in Jurassic limestone, hosts a variety of calcite speleothems, and a long succession of subterranean lakes with mixed granite and carbonate sands. The lower level is occasionally flooded by a rising groundwater level, but with only scarce input of organic remains (leaves and charcoal fragments). On the quiet cave pools there are visible calcite rafts, whereas walls are locally coated with manganese deposits. In the drier upper levels, where organic input is much more subdued, moonmilk—a hydrated calcium-magnesium carbonate speleothem—can be found. Relative humidity approaches 100% and the measured mean annual cave air temperature is 14.8°C. Samples were obtained in 2014 from calcite rafts, moonmilk, manganese oxide deposits and soil (limestone and granite grains). Microclimatic conditions in the cave near the sampling sites, sample properties, physico-chemical parameters of water, and sediment composition were determined. The microbial community of this system is predominately composed of the phyla Proteobacteria, Actinobacteria, Acidobacteria, Nitrospirae, and Firmicutes. Sampling sites near the entrance of the cave and in close proximity of the underground campsite–located 500 meters deep into the cave—revealed the highest diversity as well as the highest number of human associated microorganisms. Two samples obtained in very close proximity of each other near the campsite, indicate that the human impact is localized and is not distributed freely within the system. Analysis of the abundance of bacterial and archaeal amoA genes revealed a far greater abundance of archaeal amoA genes compared to bacterial representatives. The results of this study highlight that human impact is confined to locations that are utilized as campsites and that exploration leaves little microbial trails. Furthermore, we uncovered a highly specialized microbiome, which is perfectly adapted to survive and thrive in an environment with low nutrient availability

Microbial communities in karst groundwater and their potential use for biomonitoring

The structure, diversity and dynamics of microbial communities from a swallow hole draining agricultural land and two connected karst springs (Switzerland) were studied using molecular microbiological methods and related to hydrological and physicochemical parameters. Storm responses and an annual hydrological cycle were monitored to determine the short- and long-term variability, respectively, of bacterial communities. Statistical analysis of bacterial genetic fingerprints (16S rDNA PCR-DGGE) of spring water samples revealed several clusters that corresponded well with different levels of the allochthonous swallow hole contribution. Microbial communities in spring water samples highly affected by the swallow hole showed low similarities among them, reflecting the high temporal variability of the bacterial communities infiltrating at the swallow hole. Conversely, high similarities among samples with low allochthonous contribution provided evidence for a stable autochthonous endokarst microbial community. Three spring samples, representative for low, medium and high swallow hole contribution, were analysed by cloning/sequencing in order to identify the major bacterial groups in the communities. The autochthonous endokarst microbial community was mainly characterized of δ-Proteobacteria, Acidobacteria and Nitrospira species. A high percentage of unknown sequences suggested further that many karst aquifer bacteria are still undiscovered. Finally, the potential use of groundwater biomonitoring using microbial communities is discussed

Sulphuric acid geofluid contribution on thermal carbonate coastal springs (Italy)

Hypogenic caves, developed by sulphuric acid speleogenesis, are known all over the world among which the Santa Cesarea Terme caves have been included. They are four submerged caves, located along a coastal carbonate sector in Southern Italy and hosting the outflow of coastal springs of thermal mixed waters (from 21 to 33 °C). These waters derive from the mixing of three water end members: the fresh pure groundwater of a wide karstic aquifer, the deep sulphur thermal water and the seawater. This cave system represents an almost unique case of hypogenic sea caves in carbonate environment. The thermal mixed waters have a different effect on the surrounding rocks of the caves, influencing the sulphuric acid speleogenetic process within the whole cave system. To understand the complex and overlapping natural processes acting on the development of these coastal caves, a multidisciplinary study has been carried out. This study has integrated all the data resulting from different methods and technologies, merging morphology, structural geology, hydrogeology, hydrogeochemistry and mineralogy. This multidisciplinary study has allowed to define the main geochemical processes acting within these caves, including the cave development and the formation of the mineral concretions. After the introduction of H2S in the thermal waters, formed by the reduction of sulphates in the sedimentary deposits crossed at depth in the offshore, the oxidation occurs within the caves, producing sulphuric acid. Favoured by upwelling deep-seated thermal flows, this acid dissolves the limestone, with condensation corrosion process that involve replacement of limestone rock with gypsum. This process has resulted to be more active and remarkable within the Gattulla Cave, one of the Santa Cesarea Terme sea caves