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Kinetics of thiosulfate oxidation, product and intermediate formation, and 34S fractionation, were studied for the members of Alphaproteobacteria Paracoccus sp. SMMA5 and Mesorhizobium thiogangeticum SJTT, the Betaproteobacteria member Pusillimonas ginsengisoli SBO3, and the Acidithiobacillia member Thermithiobacillus sp. SMMA2, during chemolithoautotrophic growth in minimal salts media supplemented with 20 mM thiosulfate. The two Alphaproteobacteria oxidized thiosulfate directly to sulfate, progressively enriching the end-product with 34S; Δ34Sthiosulfate-sulfate values recorded at the end of the two processes (when no thiosulfate was oxidized any further) were −2.9‰ and −3.5‰, respectively. Pusillimonas ginsengisoli SBO3 and Thermithiobacillus sp. SMMA2, on the other hand, oxidized thiosulfate to sulfate via tetrathionate intermediate formation, with progressive 34S enrichment in the end-product sulfate throughout the incubation period; Δ34Sthiosulfate-sulfate, at the end of the two processes (when no further oxidation took place), reached −3.5‰ and −3.8‰, respectively. Based on similar 34S fractionation patterns recorded previously during thiosulfate oxidation by strains of Paracoccus pantotrophus, Advenella kashmirensis and Hydrogenovibrio crunogenus, it was concluded that progressive reverse fractionation, enriching the end-product sulfate with 34S, could be a characteristic signature of bacterial thiosulfate oxidation.

Marine sedimentation rate and bottom-water O2 concentration control the remineralization/sequestration of organic carbon across continental margins; but whether/how they shape microbiome architecture (the ultimate effector of all biogeochemical phenomena), across shelf/slope sediments, is unknown. Here we reveal distinct microbiome structures and functions, amidst comparable pore-fluid chemistries, along ~3 m sediment-horizons underlying the seasonal (shallow coastal) and perennial (deep sea) oxygen minimum zones (OMZs) of the Arabian Sea, situated across the western-Indian margin (water-depths: 31 m and, 530 and 580 m, respectively). Along the perennial- and seasonal-OMZ sediment-cores microbial communities were predominated by Gammaproteobacteria/Alphaproteobacteria and Euryarchaeota/Firmicutes respectively. As a perennial-OMZ signature, a cryptic methane production-consumption cycle was found to operate near the sediment-surface; overall diversity, as well as the relative abundances of simple-fatty-acids-requiring anaerobes (methanogens, anaerobic methane-oxidizers, sulfate-reducers and acetogens), peaked in the topmost sediment-layer and then declined via synchronized fluctuations until the sulfate-methane transition zone was reached. The entire microbiome profile was reverse in the seasonal-OMZ sediment-horizon. We discerned that in the perennial-OMZ sediments organic carbon deposited was higher in concentration, and marine components-rich, so it potentially degraded readily to simple fatty acids; lower sedimentation rate afforded higher O2 exposure time for organic matter degradation despite perennial hypoxia in the bottom-water; thus, the resultant abundance of reduced metabolites sustained multiple inter-competing microbial processes in the upper sediment-layers. Remarkably, the whole geomicrobial scenario was opposite in the sediments of the seasonal/shallow-water OMZ. Our findings create a microbiological baseline for understanding carbon-sulfur cycling across distinct marine depositional settings and water-colum n oxygenation regimes.

Geomicrobiology of sulfur–boron-dominated, neutral-pH hydrothermal systems was revealed in a Trans-Himalayan spring named Lotus Pond, located at 4436 m, in Puga Valley, Ladakh (India), where water boils at 85°C. Water sampled along Lotus Pond’s outflow (vent to an adjacent river called Rulang), representing an 85–14°C gradient, had high microbial diversity and boron/chloride/sodium/sulfate/thiosulfate concentration; potassium/silicon/sulfide/sulfite was moderately abundant, whereas cesium/lithium small but definite. Majority of the bacterial genera identified in the 85–72°C samples have no laboratory-growth reported at >45°C, and some of those mesophiles were culturable. Sulfur-species concentration and isotope-ratio along the hydrothermal gradient, together with the distribution of genera having sulfur-oxidizing members, indicated chemolithotrophic activities in the 85–72°C sites. While biodiversity increased in the vent-to-river trajectory all-day, maximum rise was invariably between the vent (85–81°C) and the 78–72°C site; below 72°C, diversity increased gradually. Biodiversity of the vent-water exhibited diurnal fluxes relatable to the sub-surface-processes-driven temporal fluxes in physicochemical properties of the discharge. Snow-melts infiltrating (via tectonic faults) the ~160°C geothermal reservoir located within the breccia, at ~450 m depth, apparently transport mesophilic microbes into the thermal waters. As these micro-organisms emanate with the vent-water, some remain alive, illustrating that natural bacterial populations are more heat-resilient than their laboratory counterparts.

Here we present high-resolution biogeochemical study using nitrogen/carbon isotope ratio measurement and molecular proxies from a sediment core (length = 2.9 m) collected from the center (588 mbsl; Lat: 16049.88’N and Long: 710 58.55’ E) of the oxygen minimum zone off west coast of India. The core archives the depositional record covering 1.114 to 12.025 ky BP. The concentrations of total organic carbon (TOC) and total nitrogen (TN) range from 0.7 to 4.9 wt% and 0.068 to 0.5 wt % respectively. TOC and TN show parallel trends and the TOC/TN ratio varies within a narrow range of 8 to 11.5. The δ13C values range from -20.5‰ to -21.9‰ (V-PDB). The TOC/TN and δ13C suggest typical marine organic matter source. This observation is also further supported by the n-alkane distribution pattern where the dominance of nC21 to nC24 and the absence of odd alkane dominance over even suggest predominantly marine organic source. The δ15N profile shows a steady increase from 5.7‰ at 203 cmbsf (5.5 ky BP) to 7.5‰ at 2 cmbsf (~1ky BP) suggesting gradual increase in denitrification possibly liked to reduced ventilation in the Arabian Sea, whereas, between 5.5 to 12 ky BP, the δ15N values show marked fluctuations (5.2 to 7.1‰) possibly indicating variable level of oxygenation which in turn controlled the extent of denitrification. Possible influence of diagenesis (microbial degradation of organic matter) on the δ15N values also need to be investigated for a better understanding of the water column processes.

To explore the potential role of tetrathionate in the sulfur cycle of marine sediments, population ecology of microorganisms capable of metabolizing this polythionate was revealed at 15–30 cm resolution along two, ~ 3-m-long, cores collected from 530 and 580 meters below the sea level, off India's west coast, within the oxygen minimum zone (OMZ) of the Arabian Sea. Metagenome analysis along the two sediment-cores revealed widespread occurrence of genes involved in microbial formation, oxidation, and reduction of tetrathionate; high diversity and relative-abundance was also detected for bacteria that are known to render these metabolisms in vitro. Results of slurry-incubation of the sediment-samples in thiosulfate- or tetrathionate-containing microbial growth media, data obtained via pure-culture isolation, and finally metatranscriptome analyses, corroborated the in situ functionality of tetrathionate-forming, oxidizing, and reducing microorganisms. Geochemical analyses revealed the presence of up to 11.1 µM thiosulfate along the two cores, except a few sample-sites near the sediment-surface. Thiosulfate oxidation by chemolithotrophic bacteria prevalent in situ is the apparent source of tetrathionate in this ecosystem. However, potential abiotic origin of the polythionate can neither be ruled out nor confirmed from the geochemical information currently available for this territory. Tetrathionate, in turn, can be either oxidized to sulfate (via oxidation by the chemolithotrophs present) or reduced back to thiosulfate (via respiration by native bacteria). Up to 2.01 mM sulfide present in the sediment-cores may also reduce tetrathionate abiotically to thiosulfate and elemental sulfur. As tetrathionate was not detected in situ, high microbiological and geochemical reactivity of this polythionate was hypothesized to be instrumental in its cryptic status as a central sulfur cycle intermediate.

To explore the potential role of tetrathionate in the sulfur cycle of marine sediments, the population ecology of tetrathionate-forming, oxidizing, and respiring microorganisms was revealed at 15-30 cm resolution along two, ∼3-m-long, cores collected from 530- and 580-mbsl water-depths of Arabian Sea, off India’s west coast, within the oxygen minimum zone (OMZ). Metagenome analysis along the two sediment-cores revealed widespread occurrence of the structural genes that govern these metabolisms; high diversity and relative-abundance was also detected for the bacteria known to render these processes. Slurry-incubation of the sediment-samples, pure-culture isolation, and metatranscriptome analysis, corroborated thein situfunctionality of all the three metabolic-types. Geochemical analyses revealed thiosulfate (0-11.1 μM), pyrite (0.05-1.09 wt %), iron (9232-17234 ppm) and manganese (71-172 ppm) along the two sediment-cores. Pyrites (via abiotic reaction with MnO2) and thiosulfate (via oxidation by chemolithotrophic bacteria prevalentin situ) are apparently the main sources of tetrathionate in this ecosystem. Tetrathionate, in turn, can be either converted to sulfate (via oxidation by the chemolithotrophs present) or reduced back to thiosulfate (via respiration by native bacteria); 0-2.01 mM sulfide present in the sediment-cores may also reduce tetrathionate abiotically to thiosulfate and elemental sulfur. Notably tetrathionate was not detectedin situ- high microbiological and geochemical reactivity of this polythionate is apparently instrumental in the cryptic nature of its potential role as a central sulfur cycle intermediate. Biogeochemical roles of this polythionate, albeit revealed here in the context of OMZ sediments, may well extend to the sulfur cycles of other geomicrobiologically-distinct marine sediment horizons.

Little is known about life in the boron-rich hot springs of Trans-Himalayas. Here, we explore the geomicrobiology of a 4438-m-high spring which emanates ~70 °C-water from a boratic microbialite called Shivlinga. Due to low atmospheric pressure, the vent-water is close to boiling point so can entropically destabilize biomacromolecular systems. Starting from the vent, Shivlinga’s geomicrobiology was revealed along the thermal gradients of an outflow-channel and a progressively-drying mineral matrix that has no running water; ecosystem constraints were then considered in relation to those of entropically comparable environments. The spring-water chemistry and sinter mineralogy were dominated by borates, sodium, thiosulfate, sulfate, sulfite, sulfide, bicarbonate, and other macromolecule-stabilizing (kosmotropic) substances. Microbial diversity was high along both of the hydrothermal gradients. Bacteria, Eukarya and Archaea constituted >98%, ~1% and Shivlinga’s microbiome, respectively. Temperature constrained the biodiversity at ~50 °C and ~60 °C, but not below 46 °C. Along each thermal gradient, in the vent-to-apron trajectory, communities were dominated by Aquificae/Deinococcus-Thermus, then Chlorobi/Chloroflexi/Cyanobacteria, and finally Bacteroidetes/Proteobacteria/Firmicutes. Interestingly, sites of >45 °C were inhabited by phylogenetic relatives of taxa for which laboratory growth is not known at >45 °C. Shivlinga’s geomicrobiology highlights the possibility that the system’s kosmotrope-dominated chemistry mitigates against the biomacromolecule-disordering effects of its thermal water.

Continental shelves are the major sites for organic carbon burial and sulfate reduction. Organic carbon degradation and sulfidization along continental shelves are expected to play a significant role in global carbon-sulfur-iron (C–S–Fe) cycles. Here we report sediment pore-water chemistry and iron-sulfur speciation in two short cores SSK-42/9 and 10 collected off Malvan from the seasonally hypoxic shelf region off the west coast of India (WCI) at water depths of 30 and 13 m respectively. Concentration profiles of pore-water SO42−, NH4+, DIC, ΣHS− and depth-integrated sulfate reduction rates (JSO4) suggest a variable influence of sedimentation rates, the composition of organic matter (marine and terrestrial) and anaerobic oxidation of methane (AOM) on the pore-fluid chemistry. Chromium reducible sulfur (CRS) and organic-bound sulfur (OBS) are the detectable solid-phase sulfur species in the studied sediment cores. Sulfur content and isotope ratios of chromium reducible sulfur (CRS) and organic-bound sulfur (OBS) phases produced via HS−/Sx2− pathways (sulfidization and sulfurization) show contrasting profiles in SSK42/9 and 10, apparently influenced by the availability of reactive iron, the relative significance of early and late diagenetic processes, source of OBS (detritus of marine origin and sulfurized organic molecules) and diffusion of ΣHS− produced via AOM across the SMTZ. The marked influence of AOM-driven sulfate reduction and the diffusion of isotopically enriched hydrogen sulfide across the sulfate-methane transition zone (SMTZ) is apparent from the δ34SCRS profiles in SSK-42/10. Sediment TOC/TS ratios recorded in this study are significantly less than that of average modern marine surface sediments (2.8:1) underlying oxygenated water and this is attributed to the enhanced sulfidization and burial of iron-sulfur minerals in the seasonally hypoxic regions. Since the inner shelf of WCI experiences seasonal alternation from normoxia to hypoxia which may have a profound influence on the benthic community structure, nature, and depth of bioturbation/bioirrigation, we propose that the impact of these processes need to be studied at a significantly higher resolution for better understanding C–S–Fe biogeochemical cycle.

Biogeochemistry of oxygen minimum zone (OMZ) sediments, which are characterized by high input of labile organic matter, have crucial bearings on the benthic biota, gas and metal fluxes across the sediment-water interface, and carbon-sulfur cycling. Here we couple pore-fluid chemistry and comprehensive microbial diversity data to reveal the sedimentary carbon-sulfur cycle across a water-depth transect covering the entire thickness of eastern Arabian Sea OMZ, off the west coast of India. Geochemical data show remarkable increase in average total organic carbon content and aerial sulfate reduction rate (JSO42−) in the sediments of the OMZ center coupled with shallowing of sulfate methane transition zone and hydrogen sulfide and ammonium build–up. Total bacterial diversity, including those of complex organic matter degraders, fermentative and exoelectrogenic bacteria, and sulfate-reducers (that utilize only simple carbon compounds) were also found to be highest in the same region. The above findings indicate that higher organic carbon sequestration from the water-columns (apparently due to lower benthic consumption, biodegradation and biotransformation) and greater bioavailability of simple organic carbon compounds (apparently produced by fermetative microflora of the sediments) are instrumental in intensifying the carbon-sulfur cycle in the sediments of the OMZ center.