The aim of this study is to identify the biomineralisation processes in hot springs of North-West Euboea Island by assessing the physico-chemical parameters of the hot water, the travertine mineralogical composition and facies, and the cyanobacterial microflora. In the studied area, the main mineral phases are calcite and aragonite, creating laminated and shrub facies of travertine deposits in close association with the cyanobacterial microflora. Microscopic analysis of fresh and cultured field samples shows the presence of 81 taxa of Cyanobacteria belonging to six orders, that is, Oscillatoriales, Synechococcales, Spirulinales, Chroococcales, Nostocales and Chroococcidiopsidales with the main factors controlling biodiversity being temperature, salinity and access to sunlight. No Cyanobacteria species were identified in areas with temperatures over 65 o C. In areas with high salinity (27-37‰), the order Oscillatoriales predominates. On the other hand, in areas with high temperatures (63 o C), fewer orders were observed, usually only Synechococcales and Spirulinales. In areas with lower temperatures (37 o C), larger numbers of Cyanobacteria orders were identified. Additionally, salinity seems to regulate the presence of the Nostocales order. The combined geobiological study revealed the presence of four biomineralisation processes involving calcium carbonate minerals, that is, (i) filamentous Cyanobacteria and extracellular polymeric substances trapping calcium carbonate crystals, (ii) extracellular polymeric substances acting as a template favouring mineral precipitation for crystal nucleation, (iii) formation of calcified Cyanobacteria sheaths and (iv) alteration of calcium carbonate crystals by endolithic Cyanobacteria. The identified biomineralisation processes suggest that the formation of calcium carbonate crystals is due to the metabolic activity of Cyanobacteria, or that the Cyanobacteria favour the deposition or the alteration of already existing crystals. The combination of F I G U R E 1 Geological map NW of Euboea (TF = Telethrio Fault, IF = Ilia Fault, AF = Aedipsos Fault and LFS = Lichades Fault System; modified after Kanellopoulos et al., 2020). The sampling sites Ilios, Casino, EOT and Platania are locations at Aedipsos.

Cyanobacteria are considered to be among the first microorganisms to settle in hot springs where they form a favourable environment for further biological establishment. Nevertheless, the exact pioneer species and how early they start participating in the biomineralisation processes remain unknown. The aim of the present study was twofold, that is, to identify the pioneer Cyanobacteria in hot springs (i.e. Aedipsos area, Greece) and to record their early biomineralisation processes. The in situ experimental approach included the setup of sterile glass and/or plexiglass slides in several locations to facilitate colonisation by Cyanobacteria, and removal of slides for study after 48-202 h. Synechococcales (37%) and Oscillatoriales (33%) were the dominant orders, followed by Chroococcales (15%) and Spirulinales (11%); whereas Chroococcidiopsidales (4%) was found only in a few sites. The order Nostocales was not observed at the early stages of colonisation although it was present in mature stages. Fortythree species of Cyanobacteria were identified as pioneer microorganisms, with Spirulina subtilissima being the most frequently found. The most common pioneers were multicellular filamentous Cyanobacteria, that is, organisms with a large surface area able to form significant amounts of extracellular polymeric substances. Among the pioneers, thermophilic species of Cyanobacteria were typical such as Chroococcidiopsis thermalis, Chroococcus thermalis, Leptolyngbya thermalis, S. subtilissima and Symploca thermalis, as well as typical limestone substrate species such as Chroococcus lithophilus and Leptolyngbya laminosa. Temperature seems to affect biodiversity. Also, pioneers were found to contribute to the biomineralisation processes from their first appearance. In the studied samples, three biomineralisation processes were identified, that is, (i) calcification of cyanobacterial sheaths, (ii) trapping of carbonate crystals on a crystal retention lattice formed by extracellular polymeric substances and filaments and (iii) trapping and confinement of carbonate crystals around filamentous Cyanobacteria.

The study of microbial mats in extreme environments is of high scientific interest from geological, ecological, and geomicrobiological aspects. These mats represent multilayer bio-structures where each taxonomic group dominates a specific vertical layering distribution resulting from its growth and metabolic activity. In the present study, microbial mats in a hot spring environment from Aedipsos (Euboea Island, Greece) resulting in the creation of thermogenic travertine, were studied through an interdisciplinary approach. The mineralogical composition was determined by optical microscopy, XRD, and SEM-EDS microanalysis, and the identification of Cyanobacteria was made primarily on morphological characteristics. The main mineral phase in the studied samples is calcite and, to a less extent, aragonite, with several trace elements in the mineral-chemistry composition, i.e., up to 1.93 wt. % MgO, up to 0.52 wt. % SrO, up to 0.44 wt. % Na2O, up to 0.17 wt. % K2O, and up to 3.99 wt. % SO3. The dominant facies are lamination and shrubs, which are the most common among the facies of thermogenic travertines of the area. Several layers were identified, (i) a top mainly abiotic layer consisting of calcium carbonate micritic crystals, (ii) a second biotic layer–the Cyanobacteria layer, dominated by the species Leptolyngbya perforans, (iii) a third biotic layer where Leptolyngbya perforans, Chloroflexus and other bacteria occur, and (iv) a deeper abiotic part with several layers where no photosynthetic microorganisms occur. In the upper layers, nineteen (19) species of Cyanobacteria were identified, classified in the orders Chroococcales (37%), Synechococcales (31%), Oscillatoriales (16%), and Spirulinales (6%). Among the identified Cyanobacteria, there are typical thermophilic and limestone substrate species. These Cyanobacteria are found to participate in the biomineralization and biologically-influenced processes, i.e., (i) filamentous Cyanobacteria are trapping calcium carbonate crystals, and diatoms, (ii) extracellular polymeric substances (EPS) create crystal retention lattice contributing to the biomineralization process, and (iii) filamentous sheaths of Cyanobacteria are calcified, resulting in the creation of calcium carbonate tubes.

Caves with hot springs and speleothem deposits are infrequent environments of high scientific interest due to their unique environmental conditions. The selected site is a small open cave with a hot spring and stalactites in the Aedipsos area (NW Euboea Island, Greece), which was studied through an interdisciplinary approach. The mineralogical composition of the speleothems was determined by optical microscopy, XRD, and SEM-EDS microanalysis, and identification of the Cyanobacteria species was made based on morphological characteristics. The main mineral phase in the studied samples is calcite, with several trace elements (i.e., up to 0.48 wt.% Na2O, up to 0.73 wt.% MgO, up to 4.19 wt.% SO3, up to 0.16 wt.% SrO and up to 2.21 wt.% Yb2O3) in the mineral-chemistry composition. The dominant facies are lamination and shrubs, which are the most common among the facies of the thermogenic travertines of the area. Based on the studied stalactites, twenty-nine different Cyanobacteria species were identified, belonging to the following orders: Synechococcales (28%), Oscillatoriales (27%), Chroococcales (21%) and Nostocales (21%), and Spirulinales (3%). Among them, thermophilic species (Spirulina subtilissima) and limestone substrate species (Chroococcus lithophilus, Leptolyngbya perforans, and Leptolyngbya ercegovicii) were identified. The identified Cyanobacteria were found to participate in biomineralization processes. The most characteristic biomineralization activity is made by the endolithic Cyanobacteria destroying calcite crystals in the outer layer. In a few cases, calcified cyanobacterial sheaths were detected. The presence of filamentous Cyanobacteria, along with extracellular polymeric substance (EPS), creates a dense net resulting in the retention of calcium carbonate crystals.