Our understanding of the icy-habitat microbiome is likely limited by a lack of reliable data on microorganisms inhabiting underground ice that has accumulated inside caves. To characterize how environmental variation impacts cave ice microbial community structure, we determined the composition of total and potentially active bacterial communities along a 13,000-year-old ice core from Scarisoara cave (Romania) through 16S rRNA gene Illumina sequencing. An average of 2,546 prokaryotic gDNA operational taxonomic units (OTUs) and 585 cDNA OTUs were identified across the perennial cave ice block and analyzed in relation to the geochemical composition of ice layers. The total microbial community and the putative active fraction displayed dissimilar taxa profiles. The ice-contained microbiome was dominated by Actinobacteria with a variable representation of Proteobacteria, while the putative active microbial community was equally shared between Proteobacteria and Firmicutes. Accordingly, a major presence of Cryobacterium, Lysinomonas, Pedobacter , and Aeromicrobium phylotypes homologous to psychrotrophic and psychrophilic bacteria from various cold environments were noted in the total community, while the prevalent putative active bacteria belonged to Clostridium, Pseudomonas, Janthinobacterium, Stenotrophomonas , and Massilia genera. Variation in the microbial cell density of ice strata with the dissolved organic carbon (DOC) content and the strong correlation of DOC and silicon concentrations revealed a major impact of depositional processes on microbial abundance throughout the ice block. Post-depositional processes appeared to occur mostly during the 4,000–7,000 years BP interval. A major bacterial composition shift was observed in 4,500–5,000-year-old ice, leading to a high representation of Beta- and Deltaproteobacteria in the potentially active community in response to the increased concentrations of DOC and major chemical elements. Estimated metabolic rates suggested the presence of a viable microbial community within the cave ice block, characterized by a maintenance metabolism in most strata and growth capacity in those ice deposits with high microbial abundance and DOC content. This first survey of microbial distribution in perennial cave ice formed since the Last Glacial period revealed a complex potentially active community, highlighting major shifts in community composition associated with geochemical changes that took place during climatic events that occurred about 5,000 years ago, with putative formation of photosynthetic biofilms.

BackgroundWe present herein our results regarding the accumulation of four heavy metals (copper, cadmium, lead, and zinc) in four aquatic species plants (Ceratophyllum demersum, Potamogeton pectinatus, Potamogeton lucens, Potamogeton perfoliatus) collected from the Danube River, South-Western part of Romania and their possible use as indicators of aquatic ecosystems pollution with heavy metals.MethodsElements concentration from the vegetal material was determined through Inductively Coupled Plasma – Mass Spectrometry.ResultsThe species were chosen based on their previous use as bioindicators in aquatic ecosystems and due to the fact they are one of the most frequent aquatic plant species of the Danube River ecosystems within the Iron Gates Natural Park. Highest amounts are recorded for Ceratophyllum demersum (3.52 μg/g for Cd; 22.71 μg/g for Cu; 20.06 μg/g for Pb; 104.23 μg/g for Zn). Among the Potamogeton species, the highest amounts of heavy metals are recorded in Potamogeton perfoliatus (1.88 μg/g for Cd; 13.14 μg/g for Cu; 13.32 μg/g for Pb; 57.96 μg/g for Zn). The sequence for the bioconcentration factors (BCFs) calculated in order to describe the accumulation of the four metals is Cd >> Zn > Pb > Cu. Increase of the zinc concentration determines an increase of the cadmium concentration (Spearman rho=0.40, p=0.02).ConclusionsDespite the low ambiental levels of heavy metals, the four aquatic plants have the ability to accumulate significant amounts, which make them useful as biological indicators. BCF value for Ceratophyllum demersum indicated this species as a cadmium hyperaccumulator.

Abstract. We present here the results of a 4-year environmental monitoring program at Ascunsȃ Cave (southwestern Romania) designed to help us understand how climate information is transferred through the karst system and archived by speleothems. The air temperature inside the cave is around 7 • C, with slight differences between the upper and lower parts of the main passage. CO 2 concentrations in cave air have a seasonal signal, with summer minima and winter maxima. These might indicate the existence of an organic matter reservoir deep within the epikarst that continues to decompose over the winter, and CO 2 concentrations are possibly modulated by seasonal differences in cave ventilation.The maximum values of CO 2 show a rise after the summer of 2014, from around 2000 to about 3500 ppm, following a rise in surface temperature. Using two newly designed types of water-air equilibrators, we were able to determine the concentration of CO 2 dissolved in drip water by measuring its concentration in the equilibrator headspace and then using Henry's law to calculate its concentration in water. This method opens the possibility of continuous data logging using infrared technology, without the need for costly and less reliable chemical determinations. The local meteoric water line (δ 2 H = 7.7 δ 18 O + 10.1), constructed using monthly aggregated rainfall samples, is similar to the global one, revealing the Atlantic as the strongly dominant vapor source. The deuterium excess values, as high as 17 ‰, indicate that precipitation has an important evaporative component, possibly given by moisture recycling over the European continent. The variability of stable isotopes in drip water is similar at all points inside the cave, suggesting that the monitored drip sites are draining a homogenous reservoir. Drip rates, as well as stable isotopes, indicate that the transfer time of water from the surface is on the order of a few days.

ABSTRACT. In the Cerna Valley basin, located southwest of the Southern Carpathians and upstream from the confluence of Cerna with Belareca, an aquifer complex has developed. The basin is strongly influenced by hydrogeothermal phenomena, acting within two major geological structures, the Cerna Syncline and the Cerna Graben. The complex consists mainly of Jurassic and Cretaceous carbonate rocks, as well as the upper part of the Cerna Granite that is highly fractured, tectonically sunken into the graben. The geothermal investigations have shown the existence of some areas with values of the geothermal gradient falling into the 110-200ºC/km interval, and temperatures of 13.8-16ºC at the depth of 30 m. The zone with the maximal flux intensity is situated between the Băile Herculane railway station and the Crucea Ghizelei Well, an area where 24 sources (10 wells and 14 springs) are known. The geothermal anomaly is also extended to the south (Topleţ), north (Mehadia) and NE (Piatra Puşcată), a fact, which is stressed by the existence of hypothermal springs with low mineralization. The physical-chemical parameters of the sources show a strong N-S variability. At the entire thermo-mineral reservoir scale, the temperature of the water sources, the total mineralization, and the H 2 S quantity are increasing from the north to the south, and the pH and natural radioactivity are following the same trend.

The purpose of this study was to examine the sensitivity, in a field situation, of the hyporheic fauna to pollution by heavy metals and also to test the use of oxidative stress enzymes produced by this fauna as a sensitive indicator of oxidative stress generated by chemical contamination. This was done by surveying the patterns of distribution, structure, and composition of hyporheic invertebrate communities in one of the most polluted rivers in Romania. Twelve permanent sampling stations with differing water qualities were established along a 180 km transect of the Arieş River. Data on hyporheic invertebrate abundance and richness, chemistry of the surface and hyporheic water and interstitial suspended particles were analyzed via multifactorial analyses. In the downstream, more polluted stations, epigean species were less abundant and hyporheic communities, especially macrocrustaceans and oligochetes, became dominant. The higher levels of hyporheic invertebrate biodiversity in the moderately polluted stations compared to highly polluted, and the increase of the number of some hyporheos (especially macrocrustaceans) in the moderately polluted stations, suggested that the hyporheic fauna was more tolerant of heavy metal pollution than the surface water fauna of the area. However, the different richness and abundance of hyporheic fauna in sites of similar water chemistry suggested that additional factors, such as sediment structure are shaping the spatial distribution of hyporheic fauna. Strong correlations between superoxide dismutase (SOD) activity in pooled tissues extracts and some chemical parameters suggest that oxidative stress enzymes may prove to be sensitive indicators of chemical pollution in hyporheic zones.

This survey presents the first high-throughput characterisation of fungal distribution based on ITS2 Illumina sequencing of uncultured microbiome from a 1500 years old perennial ice deposit in Scărișoara Ice Cave, Romania. Of the total of 1 751 957 ITS2 sequences, 64% corresponded to 182 fungal operational taxonomic units, showing a low diversity, particularly in older ice strata, and a distinct temporal distribution pattern. Ascomycota was the major phylum in all ice samples, dominating the 400 and 1500 years old ice strata deposited during the cold Little Ice Age (LIA) and Dark Ages Cold Period, while Basidiomycota was mostly present in 900-years old ice formed during the Medieval Warm Period (MWP). Chytridiomycota and Mucoromycota phyla were present in recently formed and 400-years old ice, respectively. Among the 80 identified genera, Cryptococcus victoriae, commonly found in glacial habitats, was identified in all strata. A positive correlation between fungal distribution and ice conductivity, Ca, Na and Sr concentrations was observed across the ice block, with pH values trailing climate variations during LIA and MWP, respectively. Our record highlighted the presence of a complex climate and environmental-driven fungal community in perennial ice strata accumulated during the last 1500 years in Scărișoara Ice Cave.

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During a significant flood event, reversible water exchanges may occur between a karst conduit and its adjacent porous rock (frequently designated as "matrix"): while the flood pulse rises, some conduit-derived water is forced into the matrix; then, as the flood recedes, the same water flows back into the stream passage. The present note addresses such a karst setting in the Carpathian Mountains (Romania), where in addition, a usually stable flux of chloride originating in a natural saline inflow, was being mixed with a variable flow of karst freshwater. For that particular case, with the above-mentioned process of matrix storage/release from storage assumedly taking place downstream of the mixing site, two distinct chemical signatures could be noticed during a flood event: an initial depletion in the spring flow chloride flux, subsequently followed by a comparable chloride flux enrichment (the depletion and the enrichment being outlined with respect to the essentially stable chloride flux value that had been noticed to persist at the spring over a long period of flow rate recession). Concomitantly with such flood-induced fluctuations in the spring chloride flux, the spring discharge displayed, for long periods, abnormally slow variations: the latter likely indicated that the spring supply rate actual oscillations were buffered by the reversible water exchanges which took place between the karst conduit and its adjacent matrix. On the whole, these results show that conduit-matrix water exchanges could be interpreted by simple mass balance calculations that involved fluxes of a conservative tracer (the chloride ion in that particular case).