Demonstration of a multi-technique approach to assess glacial microbial populations in the field

The ability to perform microbial detection and characterization in-field at extreme environments, rather than on returned samples, has the potential to improve the efficiency, relevance and quantity of data from field campaigns. To date, few examples of this approach have been reported. Therefore, we demonstrate that the approach is feasible in subglacial environments by deploying four techniques for microbial detection: real-time polymerase chain reaction; microscopic fluorescence cell counts, adenosine triphosphate bioluminescence assay and recombinant Factor C assay (to detect lipopolysaccharide). Each technique was applied to 12 subglacial ice samples, 12 meltwater samples and two snow samples from Engabreen, Northern Norway. Using this multi-technique approach, the detected biomarker levels were as expected, being highest in debris-rich subglacial ice, moderate in glacial meltwater and low in clean ice (debris-poor) and snow. Principal component analysis was applied to the resulting dataset and could be performed in-field to rapidly aid the allocation of resources for further sample analysis. We anticipate that in-field data collection will allow for multiple rounds of sampling, analysis, interpretation and refinement within a single field campaign, resulting in the collection of larger and more appropriate datasets, ultimately with more efficient science return

Translocation of protein tyrosine phosphatase Pez/PTPD2/PTP36 to the nucleus is associated with induction of cell proliferation

Pez is a non-transmembrane tyrosine phosphatase with homology to the FERM (4.1, ezrin, radixin, moesin) family of proteins. The subcellular localisation of Pez in endothelial cells was found to be regulated by cell density and serum concentration. In confluent monolayers Pez was cytoplasmic, but in cells cultured at low density Pez was nuclear, suggesting that it is a nuclear protein in proliferating cells. This notion is supported by the loss of nuclear Pez when cells are serum-starved to induce quiescence, and the rapid return of Pez to the nucleus upon refeeding with serum to induce proliferation. Vascular endothelial cells normally exist as a quiescent confluent monolayer but become proliferative during angiogenesis or upon vascular injury. Using a 'wound' assay to mimic these events in vitro, Pez was found to be nuclear in the cells that had migrated and were proliferative at the 'wound' edge. TGFbeta, which inhibits cell proliferation but not migration, inhibited the translocation of Pez to the nucleus in the cells at the 'wound' edge, further strengthening the argument that Pez plays a role in the nucleus during cell proliferation. Together, the data presented indicate that Pez is a nuclear tyrosine phosphatase that may play a role in cell proliferation.Carol Wadham, Jennifer R. Gamble, Mathew A. Vadas and Yeesim Khew-Goodal

Continuous summer export of nitrogen-rich organic matter from the Greenland Ice Sheet inferred by ultrahigh resolution mass spectrometry

Runoff from glaciers and ice sheets has been acknowledged as a potential source of bioavailable dissolved organic matter (DOM) to downstream ecosystems. This source may become increasingly significant as glacial melt rates increase in response to future climate change. Recent work has identified significant concentrations of bioavailable carbon and iron in Greenland Ice Sheet (GrIS) runoff. The flux characteristics and export of N-rich DOM are poorly understood. Here, we employed electrospray ionization (ESI) coupled to Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to determine the elemental compositions of DOM molecules in supraglacial water and subglacial runoff from a large GrIS outlet glacier. We provide the first detailed temporal analysis of the molecular composition of DOM exported over a full melt season. We find that DOM pools in supraglacial and subglacial runoff are compositionally diverse and that N-rich material is continuously exported throughout the melt season as the snowline retreats further inland. Identification of protein-like compounds and a high proportion of N-rich DOM, accounting for 27-41% of the DOM molecules identified by ESI FT-ICR MS, may suggest a microbial provenance and high bioavailability of glacially-exported DOM to downstream microbial communities

Implementation of in-field life detection and characterisation techniques in icy environments

An emerging trend towards non-laboratory based biological and microbiological marker analysis is occurring in multiple sectors of science and industry. In the medical sector, these trends have demonstrated that conducting sample analyses away from centralised laboratories not only makes analyses quicker and more convenient (e.g. a home pregnancy test), but can offer services that are otherwise impractical (e.g. mobile laboratories to diagnose disease in the developing world). In the environmental sector, similar benefits, plus the ability to develop and test hypotheses, protocols and sampling strategies within a field campaign, are possible with in-field analyses. Icy environments in particular would benefit from in situ or in-field life detection as they are typically remote, and hence impart high logistical costs for repeated field campaigns and associated sample return with the implication that the efficiency of scientific return is poor. Unfortunately, most equipment and protocols developed for microbiological analyses in other sectors of science and industry are unsuitable for direct application to in-field use in icy environments because of poor compatibility with icy environment sample matrices and frequently inappropriate microbiological targets. Hence within this work, two hypotheses were tested: that (i) microbiological detection infield in icy environments is possible and through this (ii) unique and more efficient scientific studies can be conducted. Cont/d.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Identification and analysis of low-molecular-weight dissolved organic carbon in subglacial basal ice ecosystems by ion chromatography

Determining the concentration and composition of dissolved organic carbon (DOC) in glacial ecosystems is important for assessments of in situ microbial activity and contributions to wider biogeochemical cycles. Nonetheless, there is limited knowledge of the abundance and character of DOC in basal ice and the subglacial environment and a lack of quantitative data on low-molecular-weight (LMW) DOC components, which are believed to be highly bioavailable to microorganisms. We investigated the abundance and composition of DOC in basal ice via a molecular-level DOC analysis. Spectrofluorometry and a novel ion chromatographic method, which has been little utilized in glacial science for LMW-DOC determinations, were employed to identify and quantify the major LMW fractions (free amino acids, carbohydrates, and carboxylic acids) in basal ice from four glaciers, each with a different type of overridden material (i.e. the pre-entrainment sedimentary type such as lacustrine material or palaeosols). Basal ice from Joyce Glacier (Antarctica) was unique in that 98% of the LMW-DOC was derived from the extremely diverse free amino acid (FAA) pool, comprising 14 FAAs. LMW-DOC concentrations in basal ice were dependent on the bioavailability of the overridden organic carbon (OC), which in turn was influenced by the type of overridden material. Mean LMW-DOC concentrations in basal ice from Russell Glacier (Greenland), Finsterwalderbreen (Svalbard), and Engabreen (Norway) were low (0–417nMC), attributed to the relatively refractory nature of the OC in the overridden palaeosols and bedrock. In contrast, mean LMW-DOC concentrations were an order of magnitude higher (4430nMC) in basal ice from Joyce Glacier, a reflection of the high bioavailability of the overridden lacustrine material (> 17% of the sediment OC comprised extractable carbohydrates, a proxy for bioavailable OC). We find that the overridden material may act as a direct (via abiotic leaching) and indirect (via microbial cycling) source of DOC to the subglacial environment and provides a range of LMW-DOC compounds that may stimulate microbial activity in wet subglacial sediments

Microbial nitrogen cycling on the Greenland Ice Sheet

Nitrogen inputs and microbial nitrogen cycling were investigated along a 79 km transect into the Greenland Ice Sheet (GrIS) during the main ablation season in summer 2010. The depletion of dissolved nitrate and production of ammonium (relative to icemelt) in cryoconite holes on Leverett Glacier, within 7.5 km of the ice sheet margin, suggested microbial uptake and ammonification respectively. Positive in situ acetylene assays indicated nitrogen fixation both in a debris-rich 100 m marginal zone and up to 5.7 km upslope on Leverett Glacier (with rates up to 16.3 μmoles C<sub>2</sub>H<sub>4</sub> m<sup>−2</sup> day<sup>−1</sup>). No positive acetylene assays were detected > 5.7 km into the ablation zone of the ice sheet. Potential nitrogen fixation only occurred when concentrations of dissolved and sediment-bound inorganic nitrogen were undetectable. Estimates of nitrogen fluxes onto the transect suggest that nitrogen fixation is likely of minor importance to the overall nitrogen budget of Leverett Glacier and of negligible importance to the nitrogen budget on the main ice sheet itself. Nitrogen fixation is however potentially important as a source of nitrogen to microbial communities in the debris-rich marginal zone close to the terminus of the glacier, where nitrogen fixation may aid the colonization of subglacial and moraine-derived debris

Greenland Ice Sheet exports labile organic carbon to the Arctic oceans

Runoff from small glacier systems contains dissolved organic carbon (DOC) rich in protein-like, low molecular weight (LMW) compounds, designating glaciers as an important source of bioavailable carbon for downstream heterotrophic activity. Fluxes of DOC and particulate organic carbon (POC) exported from large Greenland catchments, however, remain unquantified, despite the Greenland Ice Sheet (GrIS) being the largest source of global glacial runoff (ca. 400 km3 yr−1). We report high and episodic fluxes of POC and DOC from a large (> 600 km2) GrIS catchment during contrasting melt seasons. POC dominates organic carbon (OC) export (70–89% on average), is sourced from the ice sheet bed, and contains a significant bioreactive component (9% carbohydrates). A major source of the “bioavailable” (free carbohydrate) LMW–DOC fraction is microbial activity on the ice sheet surface, with some further addition of LMW–DOC to meltwaters by biogeochemical processes at the ice sheet bed. The bioavailability of the exported DOC (26–53%) to downstream marine microorganisms is similar to that reported from other glacial watersheds. Annual fluxes of DOC and free carbohydrates during two melt seasons were similar, despite the approximately two-fold difference in runoff fluxes, suggesting production-limited DOC sources. POC fluxes were also insensitive to an increase in seasonal runoff volumes, indicating a supply limitation in suspended sediment in runoff. Scaled to the GrIS, the combined DOC (0.13–0.17 TgC yr−1 (±13 %)) and POC fluxes (mean = 0.36–1.52 TgC yr−1 (±14 %)) are of a similar order of magnitude to a large Arctic river system, and hence may represent an important OC source to the near-coastal North Atlantic, Greenland and Labrador seas

Physical weathering by glaciers enhances silicon mobilisation and isotopic fractionation

Glacial meltwaters export substantial quantities of dissolved and dissolvable amorphous silicon (DSi and ASi), providing an essential nutrient for downstream diatoms. Evidence suggests that glacially exported DSi is isotopically light compared to DSi in non-glaciated rivers. However, the isotopic fractionation mechanisms are not well constrained, indicating an important gap in our understanding of processes in the global Si cycle. We use rock crushing experiments to mimic subglacial physical erosion, to provide insight into subglacial isotope fractionation. Isotopically light DSi (δ30SiDSi) released following initial dissolution of freshly ground mineral surfaces (down to −2.12 ± 0.02 ‰) suggests mechanochemical reactions induce isotopic fractionation, explaining the low δ30SiDSi composition of subglacial runoff. ASi with a consistent isotopic composition is present in all mechanically weathered samples, but concentrations are elevated in samples that have undergone more intense physical grinding. These experiments illustrate the critical role of physical processes in driving isotopic fractionation and biogeochemical weathering in subglacial environments. Understanding perturbations in high latitude Si cycling under climatic change will likely depend on the response of mechanochemical weathering to increased glacial melt

Silicon isotopes in Arctic and sub-Arctic glacial meltwaters:The role of subglacial weathering in the silicon cycle

Glacial environments play an important role in high-latitude marine nutrient cycling, potentially contributing significant fluxes of silicon (Si) to the polar oceans, either as dissolved silicon (DSi) or as dissolvable amorphous silica (ASi). Silicon is a key nutrient in promoting marine primary productivity, contributingto atmosphericCO2 removal.We present the current understanding of Si cycling in glacial systems,focusingontheSiisotope(δ30Si)composition of glacial meltwaters. We combine existing glacial δ30Si data with new measurements from 20 subArctic glaciers, showing that glacial meltwaters consistently export isotopically light DSi compared with non-glacial rivers (+0.16‰ versus +1.38‰). Glacial δ30SiASi composition ranges from −0.05‰ to −0.86‰ but exhibits low seasonal variability. Silicon fluxes and δ30Si composition from glacial systems are not commonly included in global Si budgets and isotopic mass balance calculations at present. We discuss outstanding questions, including the formation mechanism of ASi and the export of glacial nutrients from fjords. Finally, we provide a contextual framework for the recent advances in our understanding of subglacial Si cycling and highlight critical research avenues for assessing potential future changes in these environments