Sulfidic Springs Research Articles

Diverse sulfur metabolisms from two subterranean sulfidic spring systems.

In sulfidic environments, microbes oxidize reduced sulfur compounds via several pathways. We used metagenomics to investigate sulfur metabolic pathways from microbial mat communities in two subterranean sulfidic streams in Lower Kane Cave, WY, USA and from Glenwood Hot Springs, CO, USA. Both unassembled and targeted recA gene assembly analyses revealed that these streams were dominated by Epsilonproteobacteria and Gammaproteobacteria, including groups related to Sulfurovum, Sulfurospirillum, Thiothrix and an epsilonproteobacterial group with no close cultured relatives. Genes encoding sulfide:quinone oxidoreductase (SQR) were abundant at all sites, but the specific SQR type and the taxonomic affiliation of each type differed between sites. The abundance of thiosulfate oxidation pathway genes (Sox) was not consistent between sites, although overall they were less abundant than SQR genes. Furthermore, the Sox pathway appeared to be incomplete in all samples. This work reveals both variations in sulfur metabolism within and between taxonomic groups found in these systems, and the presence of novel epsilonproteobacterial groups.

A new mercury-accumulating Mucor hiemalis strain EH8 from cold sulfidic spring water biofilms.

Here, we report about a unique aquatic fungus Mucor hiemalis EH8 that can remove toxic ionic mercury from water by intracellular accumulation and reduction into elemental mercury (Hg0). EH8 was isolated from a microbial biofilm grown in sulfidic‐reducing spring water sourced at a Marching's site located downhill from hop cultivation areas with a history of mercury use. A thorough biodiversity survey and mercury‐removal function analyses were undertaken in an area of about 200 km2 in Bavaria (Germany) to find the key biofilm and microbe for mercury removal. After a systematic search using metal removal assays we identified Marching spring's biofilm out of 18 different sulfidic springs' biofilms as the only one that was capable of removing ionic Hg from water. EH8 was selected, due to its molecular biological identification as the key microorganism of this biofilm with the capability of mercury removal, and cultivated as a pure culture on solid and in liquid media to produce germinating sporangiospores. They removed 99% of mercury from water within 10–48 h after initial exposure to Hg(II). Scanning electron microscopy demonstrated occurrence of intracellular mercury in germinating sporangiospores exposed to mercury. Not only associated with intracellular components, but mercury was also found to be released and deposited as metallic‐shiny nanospheres. Electron‐dispersive x‐ray analysis of such a nanosphere confirmed presence of mercury by the HgMα peak at 2.195 keV. Thus, a first aquatic eukaryotic microbe has been found that is able to grow even at low temperature under sulfur‐reducing conditions with promising performance in mercury removal to safeguard our environment from mercury pollution.

Habitat use by two extremophile, highly endemic, and critically endangered fish species (Gambusia eurystoma and Poecilia sulphuraria; Poeciliidae)

Abstract Effective conservation measures for endangered species require basic knowledge of habitat use and critical environmental variables influencing the species' occurrence. However, setting priorities may be difficult when multiple endangered species inhabit the same area and differ in habitat use. This study characterized the physical and chemical environment at 30 sites along a 2.5 km stretch of stream associated with the Baños del Azufre hydrogen sulphide spring complex, home to the only population of the widemouth gambusia (Gambusia eurystoma) in existence. Also present in the spring is Poecilia sulphuraria, a narrowly endemic and endangered species known from only a few populations. This study provides the most detailed report to date for physical and chemical drivers of species density in this small, extreme environment. Gambusia eurystoma were generally rare throughout the stream, and stream flow and substrate size were the best predictors of G. eurystoma densities. Conversely, P. sulphuraria were considerably more abundant, but no physical or chemical variables predicted their density among sites. Size distributions of adult P. sulphuraria were significantly influenced by stream flow and water chemistry, indicating a cost to living in greater proximity to toxic springs with high hydrogen sulphide concentrations. Overall, the results provide the first extensive report of environmental variation and factors associated with G. eurystoma densities, and indicate that conservation measures prioritizing environmental conditions for G. eurystoma will also benefit P. sulphuraria. Fish in sulphide springs provide prime examples of narrowly endemic species in desperate need of conservation, and their habitats face conservation challenges similar to those found in other springs with sympatric, endemic species in the world's arid regions. Copyright © 2016 John Wiley & Sons, Ltd.

Mechanisms Underlying Adaptation to Life in Hydrogen Sulfide-Rich Environments.

Hydrogen sulfide (H2S) is a potent toxicant interfering with oxidative phosphorylation in mitochondria and creating extreme environmental conditions in aquatic ecosystems. The mechanistic basis of adaptation to perpetual exposure to H2S remains poorly understood. We investigated evolutionarily independent lineages of livebearing fishes that have colonized and adapted to springs rich in H2S and compared their genome-wide gene expression patterns with closely related lineages from adjacent, nonsulfidic streams. Significant differences in gene expression were uncovered between all sulfidic and nonsulfidic population pairs. Variation in the number of differentially expressed genes among population pairs corresponded to differences in divergence times and rates of gene flow, which is consistent with neutral drift driving a substantial portion of gene expression variation among populations. Accordingly, there was little evidence for convergent evolution shaping large-scale gene expression patterns among independent sulfide spring populations. Nonetheless, we identified a small number of genes that was consistently differentially expressed in the same direction in all sulfidic and nonsulfidic population pairs. Functional annotation of shared differentially expressed genes indicated upregulation of genes associated with enzymatic H2S detoxification and transport of oxidized sulfur species, oxidative phosphorylation, energy metabolism, and pathways involved in responses to oxidative stress. Overall, our results suggest that modification of processes associated with H2S detoxification and toxicity likely complement each other to mediate elevated H2S tolerance in sulfide spring fishes. Our analyses allow for the development of novel hypotheses about biochemical and physiological mechanisms of adaptation to extreme environments.

Complete Genome Sequence of a Strain of Azospirillum thiophilum Isolated from a Sulfide Spring.

We report the complete, closed genome sequence and complete methylome of Azospirillum thiophilum strain BV-ST.

First Azospirillum genome from aquatic environments: Whole-genome sequence of Azospirillum thiophilum BV-ST, a novel diazotroph harboring a capacity of sulfur-chemolithotrophy from a sulfide spring

Azospirillum thiophilum BV-ST, isolated from a sulfide spring, is a novel nitrogen-fixing bacterium harboring sulfur-lithotrophy. In order to identify genetic characteristics with habitat- and metabolic features contrasting to those from terrestrial Azospirillum species, we present here the genome sequence of a novel species A. thiophilum BV-ST, with a significance of first genome report in the aquatic Azospirillum species. The genome of strain BV-ST is comprised of 7.6Mb chromosome with a GC content of 68.2%. This information will contribute to expand understandings of sulfur-oxidizer microbes that preserve inherencies as a diazotroph, and further it will provide insights into genome plasticity of the genus Azospirillum for niche specific adaptations.

Controls on stable Sr-isotope fractionation in continental barite

Barite precipitation typically occurs when barium rich fluids mix with sulfate rich fluids, however barite found in the modern continental environment suggests that biological activity can play an important role in barite formation by oxidizing sulfur and/or concentrating barium within microenvironments. These activities induce barite precipitation, and carry with them implications for studies of barite genesis. Strontium (Sr) is incorporated into the barite crystal structure during barite formation preserving a radiogenic and stable Sr-isotope signature in barite, providing information about its formation. Here we present Sr-isotope results from three artesian sulfidic springs with ongoing barite precipitation (Zodletone Spring, Oklahoma; Stinking Spring, Utah; and Doughty Springs, Colorado) to explore the controls on stable Sr-isotope fractionation during barite precipitation in a continental setting. Apparent stable Sr-isotope fractionation for all three sites ranged from −0.6‰ to ~0.0‰ similar to previously published calculated values for equilibrium conditions and measured values of synthetic barite. However, clear relationships do not exist between water and barite chemistry in the natural systems, indicating that barite does not precipitate directly from solution, but heterogeneously within diverse microenvironments created by microbial biomass or on sediment surfaces. The dynamic microenvironments in a continental setting influence the apparent stable Sr-isotope fractionation during barite precipitation because of changing saturation conditions, Sr concentration and/or precipitation of different mineral phases (e.g., celestine). In order to better understand the geochemistry of barite deposits, future work is necessary to study the controls on radiogenic and stable Sr-isotope signatures of barite in the context of the temporally and spatially dynamic nature of the continental setting.

Sulfidic spring in the gypsum karst system of Monte Conca (Italy): chemistry and microbiological evidences

Monte Conca Cave is a karst system placed in Messinian evaporites, consisting of an active cave and a resurgence located on the massif of Monte Conca, Campofranco within the "Riserva Naturale Integrale di Monte Conca". A sulfidic spring is located in the terminal gallery of the cave. To characterize the physical and chemical parameters of the Monte Conca cave and of the sulfidic spring, air temperature, relative humidity, water pH, and concentrations of dissolved sulfides, nitrates and sulfates were monitored. The high sulfide consumption rate in the sulfidic spring, evaluated by a kinetic study, suggests that biotic consumption is dominant. Moreover, snottites and filamentous floating mats, rich in sulfur and nitrate suggest a microbial activity related to the sulfur cycle. Iron content was also evaluated in water and snottites, given its involvement in microbial activity. The microbial mats could be the source of an autotrophic system in close correlation with the biological cycle of many species of living organisms found near the spring. Some of them show typical troglobitic characteristics, while the presence and abundance of others depends on the water amount. The greater abundance of taxa found close to the sulfidic spring suggests a complex food web associated with it. The monitoring lasted a year and half has highlighted the difference between chemical- physical parameters of the cave and the sulfidic spring, emphasizing its typical microenvironment.

Reduction of Energetic Demands through Modification of Body Size and Routine Metabolic Rates in Extremophile Fish.

Variation in energy availability or maintenance costs in extreme environments can exert selection for efficient energy use, and reductions in organismal energy demand can be achieved in two ways: reducing body mass or metabolic suppression. Whether long-term exposure to extreme environmental conditions drives adaptive shifts in body mass or metabolic rates remains an open question. We studied body size variation and variation in routine metabolic rates in locally adapted populations of extremophile fish (Poecilia mexicana) living in toxic, hydrogen sulfide-rich springs and caves. We quantified size distributions and routine metabolic rates in wild-caught individuals from four habitat types. Compared with ancestral populations in nonsulfidic surface habitats, extremophile populations were characterized by significant reductions in body size. Despite elevated metabolic rates in cave fish, the body size reduction precipitated in significantly reduced energy demands in all extremophile populations. Laboratory experiments on common garden-raised fish indicated that elevated routine metabolic rates in cave fish likely have a genetic basis. The results of this study indicate that adaptation to extreme environments directly impacts energy metabolism, with fish living in cave and sulfide spring environments expending less energy overall during routine metabolism.

Hydrogen sulfide can inhibit and enhance oxygenic photosynthesis in a cyanobacterium from sulfidic springs

We used microsensors to investigate the combinatory effect of hydrogen sulfide (H2 S) and light on oxygenic photosynthesis in biofilms formed by a cyanobacterium from sulfidic springs. We found that photosynthesis was both positively and negatively affected by H2 S: (i) H2 S accelerated the recovery of photosynthesis after prolonged exposure to darkness and anoxia. We suggest that this is possibly due to regulatory effects of H2 S on photosystem I components and/or on the Calvin cycle. (ii) H2 S concentrations of up to 210 μM temporarily enhanced the photosynthetic rates at low irradiance. Modelling showed that this enhancement is plausibly based on changes in the light-harvesting efficiency. (iii) Above a certain light-dependent concentration threshold H2 S also acted as an inhibitor. Intriguingly, this inhibition was not instant but occurred only after a specific time interval that decreased with increasing light intensity. That photosynthesis is most sensitive to inhibition at high light intensities suggests that H2 S inactivates an intermediate of the oxygen evolving complex that accumulates with increasing light intensity. We discuss the implications of these three effects of H2 S in the context of cyanobacterial photosynthesis under conditions with diurnally fluctuating light and H2 S concentrations, such as those occurring in microbial mats and biofilms.

Convergent changes in the trophic ecology of extremophile fish along replicated environmental gradients

Summary Divergent selection along environmental gradients connecting locally restricted extreme habitats and adjacent benign habitats can shape convergent evolution of traits involved in coping with physiochemical stressors and can drive speciation. At the same time, the presence of such stressors alters aspects of the biotic environment, including resource availability and competitive regimes. However, it remains unclear whether and how the ecology of populations occurring in both extreme and benign environments varies in a predictable fashion. We investigated the trophic ecology of live‐bearing fishes of the genus Poecilia that have independently colonised multiple springs containing toxic hydrogen sulphide in southern Mexico. Sulphide spring fish are adapted to the unique environmental conditions and are reproductively isolated from ancestral populations in adjacent non‐sulphidic habitats. We used gut content analyses to test whether colonisation of extreme habitats was accompanied by shifts of trophic resource use and expansions of trophic niche width. Furthermore, we tested whether dietary shifts were reflected in trophic morphology by comparing intestinal tract lengths among populations using both wild‐caught and common garden‐raised individuals. Gut content analyses revealed that fish inhabiting toxic springs expanded their trophic niche width and changed their dietary resource use from detritus and algae to sulphide bacteria and invertebrates. This dietary shift was paralleled by changes in intestinal tract morphology, whereby sulphide spring fish had shorter intestines than fish from adjacent non‐sulphidic habitats. Analysis of common garden‐raised fish indicated that morphological differences between sulphidic and non‐sulphidic populations are at least in part due to genetic differentiation. Both patterns of trophic resource use and differentiation in trophic morphology were consistent across replicated pairs of sulphidic and non‐sulphidic populations, although the magnitude of differentiation varied among river drainages. Our results suggest that colonisation of and adaptation to sulphide springs in southern Mexico was paralleled by convergent changes in trophic ecology. This highlights the complexity of environmental gradients and the necessity of considering multiple sources of selection when studying the evolution of complex phenotypes.

Anoxygenic photosynthesis controls oxygenic photosynthesis in a cyanobacterium from a sulfidic spring.

Before the Earth's complete oxygenation (0.58 to 0.55 billion years [Ga] ago), the photic zone of the Proterozoic oceans was probably redox stratified, with a slightly aerobic, nutrient-limited upper layer above a light-limited layer that tended toward euxinia. In such oceans, cyanobacteria capable of both oxygenic and sulfide-driven anoxygenic photosynthesis played a fundamental role in the global carbon, oxygen, and sulfur cycle. We have isolated a cyanobacterium, Pseudanabaena strain FS39, in which this versatility is still conserved, and we show that the transition between the two photosynthetic modes follows a surprisingly simple kinetic regulation controlled by this organism's affinity for H2S. Specifically, oxygenic photosynthesis is performed in addition to anoxygenic photosynthesis only when H2S becomes limiting and its concentration decreases below a threshold that increases predictably with the available ambient light. The carbon-based growth rates during oxygenic and anoxygenic photosynthesis were similar. However, Pseudanabaena FS39 additionally assimilated NO3 (-) during anoxygenic photosynthesis. Thus, the transition between anoxygenic and oxygenic photosynthesis was accompanied by a shift of the C/N ratio of the total bulk biomass. These mechanisms offer new insights into the way in which, despite nutrient limitation in the oxic photic zone in the mid-Proterozoic oceans, versatile cyanobacteria might have promoted oxygenic photosynthesis and total primary productivity, a key step that enabled the complete oxygenation of our planet and the subsequent diversification of life.

Assessment of the physical environment of epigean invertebrates in a unique habitat: the case of a karst sulfidic spring, Slovenia

AbstractA well‐fissured and faulted karst area at the Žveplenica sulfidic spring created suitable conditions for epigean copepods. Their presence in sulfidic water is a rare phenomenon. To understand the conditions related to sulfidic habitats in the region, two additional springs were analysed: the Studenec karst spring and the Sovra artesian borehole, which contains dissolved sulfide. Water temperature, chemistry and the stable isotopes of oxygen and hydrogen show that the Sovra borehole water has a different origin and history to that of the Žveplenica and Studenec springs water, which is derived from local precipitation. The low slope of the δ18O–δ2H regression line for the Žveplenica spring indicates a mixing of local precipitation water with geothermal sulfidic water that may explain the presence of live copepods. These harpacticoid and cyclopoid copepods must have survived anoxia and high sulfide for some time. The mean transit time for Žveplenica spring cannot be defined precisely from the available data, although the degree of attenuation of the isotopic signal indicates mean transit times of about 1·0 to 1·5 years. Sites similar to Žveplenica spring should be carefully analysed because not only do they provide insights into biodiversity but they also represent an ecotone between the anoxic/oxic and high/low sulfide of these karst waters. Copepods in similar geological settings can be used as additional parameters to evaluate conditions in subsurface sulfidic habitats and their spatial connectivity and responses to hydrological events. Copyright © 2014 John Wiley & Sons, Ltd.

Candidatus ‘Chloroploca asiatica’ gen. nov., sp. nov., a new mesophilic filamentous anoxygenic phototrophic bacterium

Five phylogenetically similar monocultures of mesophilic filamentous anoxygenic phototrophic bacteria (FAPB) were isolated from microbial mats of low-mineral (5–28 g/L) alkaline lakes in Buryat Republic, Transbaikalia and Mongolia, as well as from biofilms of an alkaline sulfide spring (3 g/L) of the Umhei hydrothermal system (Buryat Republic). New isolates were characterized by short trichomes (15–30 μm long and ∼1 μm in diameter), straight, curved, or wavy, surrounded by a thin iron-sorbing mucous sheath. Gliding motion of the trichomes was not observed. The trichomes formed bunches consisting of several filaments. Trichomes multiply by the separation of short fragments or single cells from the parental trichome. The cells in the filaments were elongated; they contained chlorosomes, gas vesicles, poly-β-hydroxybutyrate granules, and small polyphosphate inclusions. Bacteria contained bacteriochlorophylls c and a and γ-carotene. Absorption maxima of the pigments in the cells were observed at 462, (shoulder at 515), 742, 805, and 863 nm. The organisms were strict anaerobes capable of photoautotrophic growth with sulfide as an electron donor. Elemental sulfur emerged into the medium as a result of sulfide photooxidation. The organisms were tolerant to sulfide (up to 8 mM). Best growth occurred at pH 8.0, 3–15 g/L NaCl, and 1–5 g/L sodium bicarbonate. According to phylogenetic analysis, the 16S rRNA gene sequences of the FAPB isolates formed a separate cluster most closely related to the species cluster of the family Oscillochloridaceae, suborder Chloroflexinae, order Chloroflexales, class Chloroflexi. The differences with the closest 16S rRNA gene sequences of the known FAPB were 9–10%. The formal description of a new taxon, Candidatus’ Chloroploca asiatica’ gen. nov., sp. nov., is provided.

Patterns of Macroinvertebrate and Fish Diversity in Freshwater Sulphide Springs

Extreme environments are characterised by the presence of physicochemical stressors and provide unique study systems to address problems in evolutionary ecology research. Sulphide springs provide an example of extreme freshwater environments; because hydrogen sulphide’s adverse physiological effects induce mortality in metazoans even at micromolar concentrations. Sulphide springs occur worldwide, but while microbial communities in sulphide springs have received broad attention, little is known about macroinvertebrates and fish inhabiting these toxic environments. We reviewed qualitative occurrence records of sulphide spring faunas on a global scale and present a quantitative case study comparing diversity patterns in sulphidic and adjacent non-sulphidic habitats across replicated river drainages in Southern Mexico. While detailed studies in most regions of the world remain scarce, available data suggests that sulphide spring faunas are characterised by low species richness. Dipterans (among macroinvertebrates) and cyprinodontiforms (among fishes) appear to dominate the communities in these habitats. At least in fish, there is evidence for the presence of highly endemic species and populations exclusively inhabiting sulphide springs. We provide a detailed discussion of traits that might predispose certain taxonomic groups to colonize sulphide springs, how colonizers subsequently adapt to cope with sulphide toxicity, and how adaptation may be linked to speciation processes.

Biogeographic congruency among bacterial communities from terrestrial sulfidic springs.

Terrestrial sulfidic springs support diverse microbial communities by serving as stable conduits for geochemically diverse and nutrient-rich subsurface waters. Microorganisms that colonize terrestrial springs likely originate from groundwater, but may also be sourced from the surface. As such, the biogeographic distribution of microbial communities inhabiting sulfidic springs should be controlled by a combination of spring geochemistry and surface and subsurface transport mechanisms, and not necessarily geographic proximity to other springs. We examined the bacterial diversity of seven springs to test the hypothesis that occurrence of taxonomically similar microbes, important to the sulfur cycle, at each spring is controlled by geochemistry. Complementary Sanger sequencing and 454 pyrosequencing of 16S rRNA genes retrieved five proteobacterial classes, and Bacteroidetes, Chlorobi, Chloroflexi, and Firmicutes phyla from all springs, which suggested the potential for a core sulfidic spring microbiome. Among the putative sulfide-oxidizing groups (Epsilonproteobacteria and Gammaproteobacteria), up to 83% of the sequences from geochemically similar springs clustered together. Abundant populations of Hydrogenimonas-like or Sulfurovum-like spp. (Epsilonproteobacteria) occurred with abundant Thiothrix and Thiofaba spp. (Gammaproteobacteria), but Arcobacter-like and Sulfurimonas spp. (Epsilonproteobacteria) occurred with less abundant gammaproteobacterial populations. These distribution patterns confirmed that geochemistry rather than biogeography regulates bacterial dominance at each spring. Potential biogeographic controls were related to paleogeologic sedimentation patterns that could control long-term microbial transport mechanisms that link surface and subsurface environments. Knowing the composition of a core sulfidic spring microbial community could provide a way to monitor diversity changes if a system is threatened by anthropogenic processes or climate change.

Variation in Melanism and Female Preference in Proximate but Ecologically Distinct Environments

AbstractVariation in pigmentation traits is an effective window to evolutionary processes due to their importance for survival and reproduction. In particular, one of the leading hypotheses for the maintenance of conspicuous pigmentation in natural populations is its signaling function in mate choice. Here, we demonstrate the occurrence of melanism in poeciliid fishes of the genus Poecilia that inhabits toxic, hydrogen sulfide springs in southern Mexico and the absence of melanism from closely related populations in reference habitats lacking hydrogen sulfide. Assays of female mate preference in both habitat types were used to examine whether divergence in female preference for melanism contributes to its maintenance in hydrogen sulfide springs. We found significant variation in female preferences for melanistic males. Specifically, melanistic females from the toxic spring exhibited a significant preference for melanistic males, while non‐melanistic females from the same population exhibited no preference. Females from the non‐sulfidic reference population discriminated strongly against melanistic males. Preferences of melanistic females appear to be a significant force in the maintenance of melanism in sulfidic habitats and suggest a change in preference as the divergence from non‐sulfidic ancestors. Potential polymorphism in preference within the hydrogen sulfide spring indicates that preference for melanistic males may not be environmentally controlled. Thus, a change in preference following divergence can promote the maintenance of variation in pigmentation within populations and between divergent habitats.

Coupling genetic and chemical microbiome profiling reveals heterogeneity of archaeome and bacteriome in subsurface biofilms that are dominated by the same archaeal species.

Earth harbors an enormous portion of subsurface microbial life, whose microbiome flux across geographical locations remains mainly unexplored due to difficult access to samples. Here, we investigated the microbiome relatedness of subsurface biofilms of two sulfidic springs in southeast Germany that have similar physical and chemical parameters and are fed by one deep groundwater current. Due to their unique hydrogeological setting these springs provide accessible windows to subsurface biofilms dominated by the same uncultivated archaeal species, called SM1 Euryarchaeon. Comparative analysis of infrared imaging spectra demonstrated great variations in archaeal membrane composition between biofilms of the two springs, suggesting different SM1 euryarchaeal strains of the same species at both aquifer outlets. This strain variation was supported by ultrastructural and metagenomic analyses of the archaeal biofilms, which included intergenic spacer region sequencing of the rRNA gene operon. At 16S rRNA gene level, PhyloChip G3 DNA microarray detected similar biofilm communities for archaea, but site-specific communities for bacteria. Both biofilms showed an enrichment of different deltaproteobacterial operational taxonomic units, whose families were, however, congruent as were their lipid spectra. Consequently, the function of the major proportion of the bacteriome appeared to be conserved across the geographic locations studied, which was confirmed by dsrB-directed quantitative PCR. Consequently, microbiome differences of these subsurface biofilms exist at subtle nuances for archaea (strain level variation) and at higher taxonomic levels for predominant bacteria without a substantial perturbation in bacteriome function. The results of this communication provide deep insight into the dynamics of subsurface microbial life and warrant its future investigation with regard to metabolic and genomic analyses.

Hidrogeologia do norte da Serra de Chiapas, México: modelo concetual baseado na caraterização geoquímica de nascentes cársicas salobras ricas em sulfuretos

Conspicuous sulfide-rich karst springs flow from Cretaceous carbonates in northern Sierra de Chiapas, Mexico. This is a geologically complex, tropical karst area. The physical, geologic, hydrologic and chemical attributes of these springs were determined and integrated into a conceptual hydrogeologic model. A meteoric source and a recharge elevation below 1,500 m are estimated from the spring-water isotopic signature regardless of their chemical composition. Brackish spring water flows at a maximum depth of 2,000 m, as inferred from similar chemical attributes to the produced water from a nearby oil well. Oil reservoirs may be found at depths below 2,000 m. Three subsurface environments or aquifers are identified based on the B, Li+, K+ and SiO2 concentrations, spring water temperatures, and CO2 pressures. There is mixing between these aquifers. The aquifer designated Local is shallow and contains potable water vulnerable to pollution. The aquifer named Northern receives some brackish produced water. The composition of the Southern aquifer is influenced by halite dissolution enhanced at fault detachment surfaces. Epigenic speleogenesis is associated with the Local springs. In contrast, hypogenic speleogenesis is associated with the brackish sulfidic springs from the Northern and the Southern environments.

The algal community at an ecocline of a cold sulphidic spring (Sovra artesian borehole, Slovenia)

The physicochemical parameters of the sulphide spring water from a 500 m deep artesian borehole in Sovra Valley (Slovenia) are stable, with very little variation: temperature, 9.2 ± 0.0 °C; pH 7.70 ± 0.11; specific electrical conductivity, 777 ± 8 μS/cm; and dissolved oxygen, 0.19 ± 0.11 mg/l. The water from the borehole is hard, rich in sulphates, and has a notable concentration of sulphide, while it is low in chlorides. Communities of phototrophs were analysed at the oxygenic ecocline at the outflow from the borehole, and from further downstream. Caloneis tenuis, Frustulla vulgaris, Gomphonema sp., Navicula radiosa, Oscillatoria sp., and Tribonema vulgare successfully thrive exclusively at the outflow from the borehole under sub-optimum conditions for the majority of phototrophs. Downstream from the borehole, where the hydrogen sulphide degasses, autotrophic biofilms were dominated by different diatoms. The Bray–Curtis similarity index confirms the distinctive phototrophic communities along the oxygenic ecocline downstream from the sulphidic spring. The sulphidic ecocline downstream from the spring is indicative of the composition of phototrophic communities, as some species appear only downstream from the spring. An ecocline downstream from the borehole of the sulphidic spring was reflected also in the bacterial indicator groups. This study brings novel insights to the limited knowledge in the field of oxygenic phototrophy related to sulphidic springs.