Autotrophy to Heterotrophy: Shift in Bacterial Functions During the Melt Season in Antarctic Cryoconite Holes.

Microbes residing in cryoconite holes (debris, water, and nutrient-rich ecosystems) on the glacier surface actively participate in carbon and nutrient cycling. Not much is known about how these communities and their functions change during the summer melt-season when intense ablation and runoff alter the influx and outflux of nutrients and microbes. Here, we use high-throughput-amplicon sequencing, predictive metabolic tools and Phenotype MicroArray techniques to track changes in bacterial communities and functions in cryoconite holes in a coastal Antarctic site and the surrounding fjord, during the summer season. The bacterial diversity in cryoconite hole meltwater was predominantly composed of heterotrophs (Proteobacteria) throughout the season. The associated functional potentials were related to heterotrophic-assimilatory and -dissimilatory pathways. Autotrophic Cyanobacterial lineages dominated the debris community at the beginning and end of summer, while heterotrophic Bacteroidota- and Proteobacteria-related phyla increased during the peak melt period. Predictive functional analyses based on taxonomy show a shift from predominantly phototrophy-related functions to heterotrophic assimilatory pathways as the melt-season progressed. This shift from autotrophic to heterotrophic communities within cryoconite holes can affect carbon drawdown and nutrient liberation from the glacier surface during the summer. In addition, the flushing out and export of cryoconite hole communities to the fjord could influence the biogeochemical dynamics of the fjord ecosystem.

Holed up, but thriving: Impact of multitrophic cryoconite communities on glacier elemental cycles.

Cryoconite holes (water and sediment-filled depressions), found on glacier surfaces worldwide, serve as reservoirs of microbes, carbon, trace elements, and nutrients, transferring these components downstream via glacier hydrological networks. Through targeted amplicon sequencing of carbon and nitrogen cycling genes, coupled with functional inference-based methods, we explore the functional diversity of these mini-ecosystems within Antarctica and the Himalayas. These regions showcase distinct environmental gradients and experience varying rates of environmental change influenced by global climatic shifts. Analysis revealed a diverse array of photosynthetic microorganisms, including Stramenopiles, Cyanobacteria, Rhizobiales, Burkholderiales, and photosynthetic purple sulfur Proteobacteria. Functional inference highlighted the high potential for carbohydrate, amino acid, and lipid metabolism in the Himalayan region, where organic carbon concentrations surpassed those in Antarctica by up to 2 orders of magnitude. Nitrogen cycling processes, including fixation, nitrification, and denitrification, are evident, with Antarctic cryoconite exhibiting a pronounced capacity for nitrogen fixation, potentially compensating for the limited nitrate concentrations in this region. Processes associated with the respiration of elemental sulfur and inorganic sulfur compounds such as sulfate, sulfite, thiosulfate, and sulfide suggest the presence of a complete sulfur cycle. The Himalayan region exhibits a higher potential for sulfur cycling, likely due to the abundant sulfate ions and sulfur-bearing minerals in this region. The capability for complete iron cycling through iron oxidation and reduction reactions was also predicted. Methanogenic archaea that produce methane during organic matter decomposition and methanotrophic bacteria that utilize methane as carbon and energy sources co-exist in the cryoconite, suggesting that these niches support the complete cycling of methane. Additionally, the presence of various microfauna suggests the existence of a complex food web. Collectively, these results indicate that cryoconite holes are self-sustaining ecosystems that drive elemental cycles on glaciers and potentially control carbon, nitrogen, sulfur, and iron exports downstream.

Metabolic activity and bioweathering properties of yeasts isolated from different supraglacial environments of Antarctica and Himalaya.

Yeasts have been frequently isolated from cold habitats, but their contribution to essential ecological processes such as the mineralization of organic matter in these environments is less known. Here, the diversity, metabolic capability, and extracellular enzyme profiles of yeasts from snow, blue ice and cryoconite hole environments from East Antarctica and cryoconite holes from a glacier in Western Himalaya were determined. Eighty-six yeast strains isolated were affiliated to the genera Glaciozyma, Goffeauzyma, Mrakia, Phenoliferia, and Rhodotorula. Variations in the abundance, diversity, physiological properties, extracellular enzyme and carbon substrate utilization patterns of the isolated yeasts, reflect the specific environmental conditions from which they were isolated. Overall, 20-90% of the yeasts across all habitat types and geographical locations produced extracellular enzymes to degrade proteins, esters, carbohydrates, pectin, cellulose, lignin, and tannin. About 10 and 29% of the yeasts also exhibited ability to solubilize rock-minerals like phosphate and silicate, respectively. Additionally, selected isolates were able to metabolize 28-93% of the carbon substrates comprising different compound classes on Biolog YT plates. Overall, the ability of yeasts to use diverse organic compounds prevalent on the glacier surface, points to their ecological significance in the decomposition of organic matter, cycling of nutrients, and in the weathering of minerals in supraglacial environments. Moreover, their wide metabolic capabilities suggest that they can colonize new niches and environments when meltwater export during the summer that enables links with surrounding ecosystems.

Microbial communities and their potential for degradation of dissolved organic carbon in cryoconite hole environments of Himalaya and Antarctica.

Cryoconite holes (cylindrical melt-holes on the glacier surface) are important hydrological and biological systems within glacial environments that support diverse microbial communities and biogeochemical processes. This study describes retrievable heterotrophic microbes in cryoconite hole water from three geographically distinct sites in Antarctica, and a Himalayan glacier, along with their potential to degrade organic compounds found in these environments. Microcosm experiments (22 days) show that 13-60% of the dissolved organic carbon in the water within cryoconite holes is bio-available to resident microbes. Biodegradation tests of organic compounds such as lactate, acetate, formate, propionate and oxalate that are present in cryoconite hole water show that microbes have good potential to metabolize the compounds tested. Substrate utilization tests on Biolog Ecoplate show that microbial communities in the Himalayan samples are able to oxidize a diverse array of organic substrates including carbohydrates, carboxylic acids, amino acids, amines/amides and polymers, while Antarctic communities generally utilized complex polymers. In addition, as determined by the extracellular enzyme activities, majority of the microbes (82%, total of 355) isolated in this study (Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria and Basidiomycota) had ability to degrade a variety of compounds such as proteins, lipids, carbohydrates, cellulose and lignin that are documented to be present within cryoconite holes. Thus, microbial communities have good potential to metabolize organic compounds found in the cryoconite hole environment, thereby influencing the water chemistry in these holes. Moreover, microbes exported downstream during melting and flushing of cryoconite holes may participate in carbon cycling processes in recipient ecosystems.

Spatial variability and possible sources of acetate and formate in the surface snow of East Antarctica.

Spatial trends of acetate (Ac-) and formate (Fo-) were determined in surface snow samples along a coastal-inland transect (180km) in the ice cap region at Princess Elizabeth Land and along a coastal transect in the Amery Ice Shelf (130km), East Antarctica. Variations in both Ac- and Fo- seem to be unrelated to the acidity of snow. Ionic balance determined for the snow samples indicate the availability of HNO3 that could undergo photolysis to produce hydroxyl radical (OH), one of the major reactants involved in oxidation reactions with organic matter. The strong positive correlations between Ac- and NO3- in snow from both regions indicate that NO3- mediated OH-oxidation of organic compounds in snow could be an important source of Ac- within the snowpack. On the other hand, negative correlation between Fo- and NO3- might indicate that sources other than OH-oxidation of organic matter may be dominant in the case of Fo-. Higher Ac- concentrations in the ice cap compared to the ice shelf correspond with long-range transport of biomass burning emissions to the ice cap region. Interaction of Ac- and Fo- with alkaline minerals could lead to their stability in the snowpack and minimize their loss from the snow surface. Resident microbial communities could also influence the budget of the carboxylic acids in snow.

Molecular Insights on Dissolved Organic Matter Transformation by Supraglacial Microbial Communities.

Snow overlays the majority of Antarctica and is an important repository of dissolved organic matter (DOM). DOM transformations by supraglacial microbes are not well understood. We use ultrahigh resolution mass spectrometry to elucidate molecular changes in snowpack DOM by in situ microbial processes (up to 55 days) in a coastal Antarctic site. Both autochthonous and allochthonous DOM is highly bioavailable and is transformed by resident microbial communities through parallel processes of degradation and synthesis. DOM thought to be of a more refractory nature, such as dissolved black carbon and carboxylic-rich alicyclic molecules, was also rapidly and extensively reworked. Microbially reworked DOM exhibits an increase in the number and magnitude of N-, S-, and P-containing formulas, is less oxygenated, and more aromatic when compared to the initial DOM. Shifts in the heteroatom composition suggest that microbial processes may be important in the cycling of not only C, but other elements such as N, S, and P. Microbial reworking also produces photoreactive compounds, with potential implications for DOM photochemistry. Refined measurements of supraglacial DOM and their cycling by microbes is critical for improving our understanding of supraglacial DOM cycling and the biogeochemical and ecological impacts of DOM export to downstream environments.

Microbial communities associated with Antarctic snow pack and their biogeochemical implications.

Snow ecosystems represent a large part of the Earth's biosphere and harbour diverse microbial communities. Despite our increased knowledge of snow microbial communities, the question remains as to their functional potential, particularly with respect to their role in adapting to and modifying the specific snow environment. In this work, we investigated the diversity and functional capabilities of microorganisms from 3 regions of East Antarctica, with respect to compounds present in snow and tested whether their functional signature reflected the snow environment. A diverse assemblage of bacteria (Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes, Deinococcus-Thermus, Planctomycetes, Verrucomicrobia), archaea (Euryarchaeota), and eukarya (Basidiomycota, Ascomycota, Cryptomycota and Rhizaria) were detected through culture-dependent and -independent methods. Although microbial communities observed in the three snow samples were distinctly different, all isolates tested produced one or more of the following enzymes: lipase, protease, amylase, ß-galactosidase, cellulase, and/or lignin modifying enzyme. This indicates that the snow pack microbes have the capacity to degrade organic compounds found in Antarctic snow (proteins, lipids, carbohydrates, lignin), thus highlighting their potential to be involved in snow chemistry.

Origin and sources of dissolved organic matter in snow on the East Antarctic ice sheet.

Polar ice sheets hold a significant pool of the world's carbon reserve and are an integral component of the global carbon cycle. Yet, organic carbon composition and cycling in these systems is least understood. Here, we use ultrahigh resolution mass spectrometry to elucidate, at an unprecedented level, molecular details of dissolved organic matter (DOM) in Antarctic snow. Tens of thousands of distinct molecular species are identified, providing clues to the nature and sources of organic carbon in Antarctica. We show that many of the identified supraglacial organic matter formulas are consistent with material from microbial sources, and terrestrial inputs of vascular plant-derived materials are likely more important sources of organic carbon to Antarctica than previously thought. Black carbon-like material apparently originating from biomass burning in South America is also present, while a smaller fraction originated from soil humics and appears to be photochemically or microbially modified. In addition to remote continental sources, we document signals of oceanic emissions of primary aerosols and secondary organic aerosol precursors. The new insights on the diversity of organic species in Antarctic snowpack reinforce the importance of studying organic carbon associated with the Earth's polar regions in the face of changing climate.

Diversity and physiology of culturable bacteria associated with a coastal Antarctic ice core.

Microbiological studies of polar ice at different depths may provide important comparisons, as they preserve records of microbial cells and past climate. In this study, we examined bacterial abundance, diversity and glaciochemical composition from three depths of an ice core from coastal Dronning Maud Land, East Antarctica. Higher bacterial abundance corresponded with high in situ sea-salt Na(+) and dust concentration, suggesting that bacteria might have been transported and deposited into ice along with dust particles and marine aerosols. Fourteen bacterial isolates belonging to the genera Methylobacterium, Brevundimonas, Paenibacillus, Bacillus and Micrococcus were retrieved. Frequent isolation of similar bacterial genera from different cold environments suggests that they possess features that enable survival and metabolism for extended periods of time at sub-zero temperatures. The highest number and diversity of recoverable bacteria was obtained from 49 m depth corresponding to 1926 AD and consisted of bacteria from 4 different genera whereas at 11 m (1989 AD) and 33 m (1953 AD) samples only species belonging to the genera Bacillus was recovered. Among the Bacillus species, Bacillus aryabhattai which has been reported only from the upper stratosphere, was isolated and is the first record from the Earth's surface. Methylobacterium was the most dominant genera at 49 m depth and its prevalence is attributable to a combination of high in situ methanesulfonate concentration, specialized metabolism and environmental hardiness of Methylobacterium. Some of the isolated bacteria were found to respire and grow using methanesulfonate, suggesting that they may utilize this substrate to sustain growth in ice. In addition, NO(3)(-) (2.93-3.69 µM), NH(4)(+) (1.45-3.90 µM) and PO(4)(3-) (0.01-0.75 µM) present in the ice could be potential sources fueling bacterial metabolism in this environment. It could be deduced from the study that variation in bacterial abundance and diversity was probably associated with the prevailing in situ conditions in ice.

Microbial preference for different size classes of organic carbon: a study from Antarctic snow.

Significance of carbon cycling in polar ecosystems is well recognized. Yet, bacteria in surface snow have received less attention in terms of their potential in carbon cycling. Here, we present results on carbon utilization by bacterial communities in three surface snow samples from Antarctica collected along a coastal to inland transect. Microcosm studies were conducted over 8 days at 5 ± 1°C to study carbon metabolism in different combinations of added low molecular weight (LMW (glucose, <1 kDa)) and high molecular weight (HMW (starch, >1 kDa)) substrates (final 20 ppm). The total organic carbon (TOC) in the snow samples decreased with time at rates ranging from non-detectable to 1.4 ppm day(-1) with rates highest in snow samples from inland region. In addition, carbon utilization studies were also carried out with bacterial isolates LH1, LH2, and LH4 belonging to the genus Cellulosimicrobium, Bacillus, and Ralstonia, respectively, isolated from the snow samples. Studies with strain LH2 in different amendments of glucose and starch showed that TOC decreased with time in all amendments at a rate of 0.9-1.5 ppm day(-1) with highest rates of 1.4-1.5 ppm day(-1) in amendments containing a higher proportion of starch. The bacterial isolates were also studied to determine their ability to utilize other LMW and HMW compounds. They utilized diverse substrates like carbohydrates, amino acids, amines, amides, complex polymers, etc., of molecular mass <100 Da, 100-500 Da, >500 Da-1 kDa, and >1 kDa preferring (up to 31 times) substrates with mass of >1 kDa than <1 kDa. The ability of bacteria in snow to utilize diverse LMW and HMW substrates indicates that they could be important in the uptake of similar compounds in snow and therefore potentially govern snow chemistry.

Organic carbon in Antarctic snow: spatial trends and possible sources.

Organic carbon records in Antarctic snow are sparse despite the fact that it is of great significance to global carbon dynamics, snow photochemistry, and air-snow exchange processes. Here, surface snow total organic carbon (TOC) along with sea-salt Na(+), dust, and microbial load of two geographically distinct traverses in East Antarctica are presented, viz. Princess Elizabeth Land (PEL, coast to 180 km inland, Indian Ocean sector) and Dronning Maud Land (DML, ∼110-300 km inland, Atlantic Ocean sector). TOC ranged from 88 ± 4 to 928 ± 21 µg L(-1) in PEL and 13 ± 1 to 345 ± 6 µg L(-1) in DML. TOC exhibited considerable spatial variation with significantly higher values in the coastal samples (p < 0.001), but regional variation was insignificant within the two transects beyond 100 km (p > 0.1). Both distance from the sea and elevation influenced TOC concentrations. TOC also showed a strong positive correlation with sea-salt Na(+) (p < 0.001). In addition to marine contribution, in situ microorganisms accounted for 365 and 320 ng carbon L(-1) in PEL and DML, respectively. Correlation with dust suggests that crustal contribution of organic carbon was marginal. Though TOC was predominantly influenced by marine sources associated with sea-spray aerosols, local microbial contributions were significant in distant locations having minimal sea-spray input.

Cobalt immobilization by manganese oxidizing bacteria from the Indian ridge system.

Co immobilization by two manganese oxidizing isolates from Carlsberg Ridge waters (CR35 and CR48) was compared with that of Mn at same molar concentrations. At a lower concentration of 10 µM, CR35 and CR48 immobilized 22 and 23 fM Co cell(-1) respectively, which was 1.4 to 2 times higher than that of Mn oxidation, while at 10 mM the immobilization was 15-69 times lower than that of Mn. Scanning electron microscope and energy dispersive X-ray analyses of intact bacterial cells grown in 1 mM Co revealed Co peaks showing extracellular binding of the metal. However, it was evident from transmission electron microscope analyses that most of the sequestered Co was bound intracellularly along the cell membrane in both the isolates. Change in morphology was one of the strategies bacteria adopted to counter metal stress. The cells grew larger and thus maintained a lower than normal surface area-volume ratio on exposure to Co to reduce the number of binding sites. An unbalanced growth with increasing Co additions was observed in the isolates. Cells attained a length of 10-18 µm at 10 mM Co which was 11-15 times the original cell length. Extensive cell rupture indicated that Co was harmful at this concentration. It is apparent that biological and optimal requirement of Mn is more than Co. Thus, these differences in the immobilization of the two metals could be driven by the differences in the requirement, cell physiology and the affinities of the isolates for the concentrations of the metals tested.

Phenotypic and molecular identification of Cellulosimicrobium cellulans isolated from Antarctic snow.

We report for the first time the isolation of Cellulosimicrobium cellulans from Antarctic snow. This strain demonstrated physiological traits that were markedly different from that of the mesophilic C. cellulans type strain DSM 43879(T). The dominant cell wall sugars in C. cellulans were glucose, galactose and mannitol whereas rhamnose was the only major sugar in the type strain. Cellular fatty acid patterns were dominated by 12-methyltetradecanoic acid (ai-C(15:0)), hexadecanoic acid (C(16:0)) and 14-methylhexadecanoic acid (ai-C(17:0)) but lacked iso fatty acids unlike the type strain. The ability of C. cellulans to survive in Antarctic snow could be due to these modified physiological properties that distinguish it from its mesophilic counterpart. Carbon utilization studies demonstrated that C. cellulans preferred complex carbon substrates over simple ones suggesting that it could play a potential role in carbon uptake in snow. Our study shows that this genus could be more cosmopolitan than hitherto thought of and is capable of living in extreme cold environments.