Jill A. Mikucki

A Contemporary Microbially Maintained Subglacial Ferrous “Ocean”

An active microbial assemblage cycles sulfur in a sulfate-rich, ancient marine brine beneath Taylor Glacier, an outlet glacier of the East Antarctic Ice Sheet, with Fe(III) serving as the terminal electron acceptor. Isotopic measurements of sulfate, water, carbonate, and ferrous iron and functional gene analyses of adenosine 5′-phosphosulfate reductase imply that a microbial consortium facilitates a catalytic sulfur cycle. These metabolic pathways result from a limited organic carbon supply because of the absence of contemporary photosynthesis, yielding a subglacial ferrous brine that is anoxic but not sulfidic. Coupled biogeochemical processes below the glacier enable subglacial microbes to grow in extended isolation, demonstrating how analogous organic-starved systems, such as Neoproterozoic oceans, accumulated Fe(II) despite the presence of an active sulfur cycle.

Bacterial Diversity Associated with Blood Falls, a Subglacial Outflow from the Taylor Glacier, Antarctica

ABSTRACT Blood Falls is the surface manifestation of brine released from below the Taylor Glacier, McMurdo Dry Valleys, Antarctica. Geochemical analyses of Blood Falls show that this brine is of a marine origin. The discovery that 74% of clones and isolates from Blood Falls share high 16S rRNA gene sequence homology with phylotypes from marine systems supports this contention. The bacterial 16S rRNA gene clone library was dominated by a phylotype that had 99% sequence identity with Thiomicrospira arctica (46% of the library), a psychrophilic marine autotrophic sulfur oxidizer. The remainder of the library contained phylotypes related to the classes Betaproteobacteria, Deltaproteobacteria, and Gammaproteobacteria and the division Bacteroidetes and included clones whose closest cultured relatives metabolize iron and sulfur compounds. These findings are consistent with the high iron and sulfate concentrations detected in Blood Falls, which are likely due to the interactions of the subglacial brine with the underlying iron-rich bedrock. Our results, together with previous reports, suggest that the brine below the Taylor Glacier hosts a viable ecosystem with microorganisms capable of growth, supported by chemical energy present in reduced iron and sulfur compounds. The metabolic and phylogenetic structure of this subglacial microbial assemblage appears to be controlled by glacier hydrology, bedrock lithology, and the preglacial ecosystem.

Metabolic activity and diversity of cryoconites in the Taylor Valley, Antarctica

[1] Metabolic activity and biogeochemical diversity within cryoconites from the Canada, Commonwealth, Howard, and Hughes glaciers in the McMurdo Dry Valleys revealed the presence of a productive microbial refuge in this polar desert ecosystem. Fluorescent in situ hybridization showed a high percentage of Cytophaga-Flavobacteria cells in cryoconite sediments (87.2%), while b-Proteobacterial cells dominated the ice overlying the sediment layer (54.2%). The biomass of bacterial cells in the sediments was also greater (4.82 mgC ml � 1 ) than that in the overlying ice (0.18 mgC ml � 1 ) and was related to bacterial productivity (on the basis of thymidine incorporation), which ranged from 36 ng Cl � 1 d � 1 in the overlying ice to 3329 ng C l � 1 d � 1 in the sediment-containing layers. Bacteria within both the sediments and overlying ice were able to actively incorporate and respire radio-labeled glucose, as well as 17 other dissolved organic carbon compounds. The cryoconites in the Taylor Valley support an active, diverse assemblage of organisms despite the fact that they may remain sealed from the atmosphere for decades. Given the density of the cryoconites in the dry valleys (� 4–6% of ablation zone surfaces), flushing of the cryoconites during warm years could provide a vital nutrient and organic carbon source to the surrounding polar desert.

WISSARD at Subglacial Lake Whillans, West Antarctica: scientific operations and initial observations

Abstract A clean hot-water drill was used to gain access to Subglacial Lake Whillans (SLW) in late January 2013 as part of the Whillans Ice Stream Subglacial Access Research Drilling (WISSARD) project. Over 3 days, we deployed an array of scientific tools through the SLW borehole: a downhole camera, a conductivity–temperature–depth (CTD) probe, a Niskin water sampler, an in situ filtration unit, three different sediment corers, a geothermal probe and a geophysical sensor string. Our observations confirm the existence of a subglacial water reservoir whose presence was previously inferred from satellite altimetry and surface geophysics. Subglacial water is about two orders of magnitude less saline than sea water (0.37–0.41 psu vs 35 psu) and two orders of magnitude more saline than pure drill meltwater (<0.002 psu). It reaches a minimum temperature of –0.55~C, consistent with depression of the freezing point by 7.019 MPa of water pressure. Subglacial water was turbid and remained turbid following filtration through 0.45 µm filters. The recovered sediment cores, which sampled down to 0.8 m below the lake bottom, contained a macroscopically structureless diamicton with shear strength between 2 and 6 kPa. Our main operational recommendation for future subglacial access through water-filled boreholes is to supply enough heat to the top of the borehole to keep it from freezing.

Thermodynamic Constraints on Microbially Mediated Processes in Lakes of the McMurdo Dry Valleys, Antarctica

Recent research has shown that the distribution of biogenic nitrogen and sulfur compounds (in particular, N 2 O, DMS and dissolved DMSO) observed in five perennially ice-covered lakes of the McMurdo Dry Valleys, Antarctica, appear to lack obvious biogeochemical explanations. This study examined the hypothesis that the distribution of these compounds resulted from thermodynamic constraints on microbially mediated processes. The thermodynamic favorableness of a number of ecologically important redox reactions in these lakes was assessed using a simplified Nernst equation and mathematically modified E h measurements. Our qualitative analysis revealed that the relative dominance of denitrification and nitrification as formation and loss processes for N 2 O was a complex pattern that was related to the redox conditions present. The results indicate that nitrification was initially the sole pathway for the formation of N 2 O with denitrification being thermodynamically unfavorable. As the redox conditions became less oxic, both denitrification and nitrification were possible. Once complete anoxia occurred, the importance of nitrification decreased considerably and the loss of N 2 O via denitrification became increasingly important. With respect to DMSO, the results indicated that DMSO was thermodynamically unfavorable as an electron acceptor for the respiration of organic material by bacteria in well-oxygenated environments, giving a partial explanation for the relatively elevated levels of DMSO found in many aquatic environments. Overall, the results of this study showed that subtle variations in the redox conditions present provided a plausible explanation for the unusual distributions of biogenic sulfur and nitrogen compounds observed in the lakes of the McMurdo Dry Valleys.

IceMole: a maneuverable probe for clean in situ analysis and sampling of subsurface ice and subglacial aquatic ecosystems

Abstract There is significant interest in sampling subglacial environments for geobiological studies, but they are difficult to access. Existing ice-drilling technologies make it cumbersome to maintain microbiologically clean access for sample acquisition and environmental stewardship of potentially fragile subglacial aquatic ecosystems. The IceMole is a maneuverable subsurface ice probe for clean in situ analysis and sampling of glacial ice and subglacial materials. The design is based on the novel concept of combining melting and mechanical propulsion. It can change melting direction by differential heating of the melting head and optional side-wall heaters. The first two prototypes were successfully tested between 2010 and 2012 on glaciers in Switzerland and Iceland. They demonstrated downward, horizontal and upward melting, as well as curve driving and dirt layer penetration. A more advanced probe is currently under development as part of the Enceladus Explorer (EnEx) project. It offers systems for obstacle avoidance, target detection, and navigation in ice. For the EnEx-IceMole, we will pay particular attention to clean protocols for the sampling of subglacial materials for biogeochemical analysis. We plan to use this probe for clean access into a unique subglacial aquatic environment at Blood Falls, Antarctica, with return of a subglacial brine sample.

Microbial sulfur transformations in sediments from Subglacial Lake Whillans

Diverse microbial assemblages inhabit subglacial aquatic environments. While few of these environments have been sampled, data reveal that subglacial organisms gain energy for growth from reduced minerals containing nitrogen, iron, and sulfur. Here we investigate the role of microbially mediated sulfur transformations in sediments from Subglacial Lake Whillans (SLW), Antarctica, by examining key genes involved in dissimilatory sulfur oxidation and reduction. The presence of sulfur transformation genes throughout the top 34 cm of SLW sediments changes with depth. SLW surficial sediments were dominated by genes related to known sulfur-oxidizing chemoautotrophs. Sequences encoding the adenosine-5′-phosphosulfate (APS) reductase gene, involved in both dissimilatory sulfate reduction and sulfur oxidation, were present in all samples and clustered into 16 distinct operational taxonomic units. The majority of APS reductase sequences (74%) clustered with known sulfur oxidizers including those within the “Sideroxydans” and Thiobacillus genera. Reverse-acting dissimilatory sulfite reductase (rDSR) and 16S rRNA gene sequences further support dominance of “Sideroxydans” and Thiobacillus phylotypes in the top 2 cm of SLW sediments. The SLW microbial community has the genetic potential for sulfate reduction which is supported by experimentally measured low rates (1.4 pmol cm-3d-1) of biologically mediated sulfate reduction and the presence of APS reductase and DSR gene sequences related to Desulfobacteraceae and Desulfotomaculum. Our results also infer the presence of sulfur oxidation, which can be a significant energetic pathway for chemosynthetic biosynthesis in SLW sediments. The water in SLW ultimately flows into the Ross Sea where intermediates from subglacial sulfur transformations can influence the flux of solutes to the Southern Ocean.

Subglacial Lake Whillans microbial biogeochemistry: a synthesis of current knowledge

Liquid water occurs below glaciers and ice sheets globally, enabling the existence of an array of aquatic microbial ecosystems. In Antarctica, large subglacial lakes are present beneath hundreds to thousands of metres of ice, and scientific interest in exploring these environments has escalated over the past decade. After years of planning, the first team of scientists and engineers cleanly accessed and retrieved pristine samples from a West Antarctic subglacial lake ecosystem in January 2013. This paper reviews the findings to date on Subglacial Lake Whillans and presents new supporting data on the carbon and energy metabolism of resident microbes. The analysis of water and sediments from the lake revealed a diverse microbial community composed of bacteria and archaea that are close relatives of species known to use reduced N, S or Fe and CH4 as energy sources. The water chemistry of Subglacial Lake Whillans was dominated by weathering products from silicate minerals with a minor influence from seawater. Contributions to water chemistry from microbial sulfide oxidation and carbonation reactions were supported by genomic data. Collectively, these results provide unequivocal evidence that subglacial environments in this region of West Antarctica host active microbial ecosystems that participate in subglacial biogeochemical cycling.

Microbial Community Structure of Subglacial Lake Whillans, West Antarctica

Subglacial Lake Whillans (SLW) is located beneath ∼800 m of ice on the Whillans Ice Stream in West Antarctica and was sampled in January of 2013, providing the first opportunity to directly examine water and sediments from an Antarctic subglacial lake. To minimize the introduction of surface contaminants to SLW during its exploration, an access borehole was created using a microbiologically clean hot water drill designed to reduce the number and viability of microorganisms in the drilling water. Analysis of 16S rRNA genes (rDNA) amplified from samples of the drilling and borehole water allowed an evaluation of the efficacy of this approach and enabled a confident assessment of the SLW ecosystem inhabitants. Based on an analysis of 16S rDNA and rRNA (i.e., reverse-transcribed rRNA molecules) data, the SLW community was found to be bacterially dominated and compositionally distinct from the assemblages identified in the drill system. The abundance of bacteria (e.g., Candidatus Nitrotoga, Sideroxydans, Thiobacillus, and Albidiferax) and archaea (Candidatus Nitrosoarchaeum) related to chemolithoautotrophs was consistent with the oxidation of reduced iron, sulfur, and nitrogen compounds having important roles as pathways for primary production in this permanently dark ecosystem. Further, the prevalence of Methylobacter in surficial lake sediments combined with the detection of methanogenic taxa in the deepest sediment horizons analyzed (34–36 cm) supported the hypothesis that methane cycling occurs beneath the West Antarctic Ice Sheet. Large ratios of rRNA to rDNA were observed for several operational taxonomic units abundant in the water column and sediments (e.g., Albidiferax, Methylobacter, Candidatus Nitrotoga, Sideroxydans, and Smithella), suggesting a potentially active role for these taxa in the SLW ecosystem. Our findings are consistent with chemosynthetic microorganisms serving as the ecological foundation in this dark subsurface environment, providing new organic matter that sustains a microbial ecosystem beneath the West Antarctic Ice Sheet.

Physiological Ecology of Microorganisms in Subglacial Lake Whillans

Subglacial microbial habitats are widespread in glaciated regions of our planet. Some of these environments have been isolated from the atmosphere and from sunlight for many thousands of years. Consequently, ecosystem processes must rely on energy gained from the oxidation of inorganic substrates or detrital organic matter. Subglacial Lake Whillans (SLW) is one of more than 400 subglacial lakes known to exist under the Antarctic ice sheet; however, little is known about microbial physiology and energetics in these systems. When it was sampled through its 800 m thick ice cover in 2013, the SLW water column was shallow (~2 m deep), oxygenated, and possessed sufficient concentrations of C, N, and P substrates to support microbial growth. Here, we use a combination of physiological assays and models to assess the energetics of microbial life in SLW. In general, SLW microorganisms grew slowly in this energy-limited environment. Heterotrophic cellular carbon turnover times, calculated from 3H-thymidine and 3H-leucine incorporation rates, were long (60 to 500 days) while cellular doubling times averaged 196 days. Inferred growth rates (average ~0.006 d−1) obtained from the same incubations were at least an order of magnitude lower than those measured in Antarctic surface lakes and oligotrophic areas of the ocean. Low growth efficiency (8%) indicated that heterotrophic populations in SLW partition a majority of their carbon demand to cellular maintenance rather than growth. Chemoautotrophic CO2-fixation exceeded heterotrophic organic C-demand by a factor of ~1.5. Aerobic respiratory activity associated with heterotrophic and chemoautotrophic metabolism surpassed the estimated supply of oxygen to SLW, implying that microbial activity could deplete the oxygenated waters, resulting in anoxia. We used thermodynamic calculations to examine the biogeochemical and energetic consequences of environmentally imposed switching between aerobic and anaerobic metabolisms in the SLW water column. Heterotrophic metabolisms utilizing acetate and formate as electron donors yielded less energy than chemolithotrophic metabolisms when calculated in terms of energy density, which supports experimental results that showed chemoautotrophic activity in excess of heterotrophic activity. The microbial communities of subglacial lake ecosystems provide important natural laboratories to study the physiological and biogeochemical behavior of microorganisms inhabiting cold, dark environments.