竹内 望

Abstract Glaciers host ecosystems comprised of biodiverse and active microbiota. Among glacial ecosystems, less is known about the ecology of ice caps since most studies focus on valley glaciers or ice sheet margins. Previously we detailed the microbiota of one such high Arctic ice cap, focusing on cryoconite as a microbe‐mineral aggregate formed by cyanobacteria. Here, we employ metabolomics at the scale of an entire ice cap to reveal the major metabolic pathways prevailing in the cryoconite of Foxfonna, central Svalbard. We reveal how geophysical and biotic processes influence the metabolomes of its resident cryoconite microbiota. We observed differences in amino acid, fatty acid, and nucleotide synthesis across the cap reflecting the influence of ice topography and the cyanobacteria within cryoconite. Ice topography influences central carbohydrate metabolism and nitrogen assimilation, whereas bacterial community structure governs lipid, nucleotide, and carotenoid biosynthesis processes. The prominence of polyamine metabolism and nitrogen assimilation highlights the importance of recycling nitrogenous nutrients. To our knowledge, this study represents the first application of metabolomics across an entire ice mass, demonstrating its utility as a tool for revealing the fundamental metabolic processes essential for sustaining life in supraglacial ecosystems experiencing profound change due to Arctic climate change‐driven mass loss.

Abstract. Cryoconite holes (CHs) are water-filled cylindrical holes with cryoconite (dark-coloured sediment) deposited at their bottoms, forming on ablatingice surfaces of glaciers and ice sheets worldwide. Because the collapse of CHs may disperse cryoconite on the ice surface, thereby decreasing theice surface albedo, accurate simulation of the temporal changes in CH depth is essential for understanding ice surface melt. We established a novelmodel that simulates the temporal changes in CH depth using heat budgets calculated independently at the ice surface and CH bottom based onhole-shaped geometry. We evaluated the model with in situ observations of the CH depths on the Qaanaaq ice cap in northwestern Greenland during the2012, 2014, and 2017 melt seasons. The model reproduced the observed depth changes and timing of CH collapse well. Although earlier models haveshown that CH depth tends to be deeper when downward shortwave radiation is intense, our sensitivity tests suggest that deeper CH tends to form whenthe diffuse component of downward shortwave radiation is dominant, whereas CHs tend to be shallower when the direct component is dominant. Inaddition, the total heat flux to the CH bottom is dominated by shortwave radiation transmitted through ice rather than that directly from theCH mouths when the CH is deeper than 0.01 m. Because the shortwave radiation transmitted through ice can reach the CH bottom regardless ofCH diameter, CH depth is unlikely to be correlated with CH diameter. The relationship is consistent with previous observationalstudies. Furthermore, the simulations highlighted that the difference in albedo between ice surface and CH bottom was a key factor for reproducingthe timing of CH collapse. It implies that lower ice surface albedo could induce CH collapse and thus cause further lowering of the albedo. Heatcomponent analysis suggests that CH depth is governed by the balance between the intensity of the diffuse component of downward shortwave radiationand the turbulent heat transfer. Therefore, these meteorological conditions may be important factors contributing to the recent surface darkening ofthe Greenland ice sheet and other glaciers via the redistribution of CHs.

Abstract Glaciers are inhabited by various cryophilic organisms ranging from single celled to multicellular, like Tardigrada (water bears). Owing to their scattered distribution, glaciers represent extremely fragmented habitats, and it remains unclear how their inhabitants survive and disperse among such isolated patches. This study investigates the biogeography of the tardigrade genus Cryoconicus, whose distribution, population stability, and interregional connectivity are examined by screening the collections from ~ 60 glaciers worldwide and by a phylogeographic analysis. We found that two Cryoconicus species occur at low densities on two Arctic glaciers in Svalbard, far from their previously reported Antarctic and Central Asian ranges. Screening of worldwide databases and DNA metabarcoding indicated that these species are absent or rare in the intermediate areas, suggesting large disjunctions in their ranges. In particular, the genetic data and multiyear resampling showed that Cryoconicus kaczmareki established a stable population on the Ebba Glacier (Svalbard), which has been isolated from its Asian core range since before the last glacial maximum. Our findings suggest that glacial invertebrates may possess wide yet largely disjunctive ranges. Interpolar- or intercontinental-scale movements of cryophilic meiofauna may occur, but migration connectivity is not sufficient to mitigate the differentiation of the local population. Revealed biogeographic patterns further demonstrate that inhabitants of extreme environments may establish isolated and highly fragmented populations that persist long term, even if at very low densities.

Introduction Microbial communities are important components of glacier and snowpack ecosystems that influence biogeochemical cycles and snow/ice melt. Recent environmental DNA surveys have revealed that chytrids dominate the fungal communities in polar and alpine snowpacks. These could be parasitic chytrids that infect snow algae as observed microscopically. However, the diversity and phylogenetic position of parasitic chytrids has not been identified due to difficulties in establishing their culture and subsequent DNA sequencing. In this study, we aimed to identify the phylogenetic positions of chytrids infecting the snow algae, Chloromonas spp., bloomed on snowpacks in Japan. Methods By linking a microscopically picked single fungal sporangium on a snow algal cell to a subsequent sequence of ribosomal marker genes, we identified three novel lineages with distinct morphologies. Results All the three lineages belonged to Mesochytriales, located within “Snow Clade 1”, a novel clade consisting of uncultured chytrids from snow-covered environments worldwide. Additionally, putative resting spores of chytrids attached to snow algal cells were observed. Discussion This suggests that chytrids may survive as resting stage in soil after snowmelt. Our study highlights the potential importance of parasitic chytrids that infect snow algal communities.

Glacier algae, which are photosynthetic microbes growing on ice, considerably reduce the surface albedo of glaciers and accelerate their melting rate. Although the growth of glacier algae can be suppressed by parasitic chytrids, the impact of chytrids on algal populations is still largely unknown. In this study, we described the morphology of the chytrid infecting the glacier alga Ancylonema nordenskioeldii and quantified the prevalence of infection in different habitats on a mountain glacier in Alaska, USA. Microscopic observations revealed three different morphological types of chytrids with distinct rhizoid shapes. Variations in the size of the sporangia were probably because of their different growth stages, indicating that they actively propagated on the glacier. The prevalence of infection did not vary among sites with different elevations but was substantially higher in cryoconite holes (20%) than on ice surfaces (4%) at all sites. This indicates that cryoconite holes are hot spots for chytrid infections of glacier algae, and the dynamics of cryoconite holes might affect the host-parasite interactions between chytrids and the glacier algae, which may in turn alter surface albedo and ice melting.

Recent studies of microbial biogeography have revealed the global distribution of cosmopolitans and dispersal of regional endemics, but little is known about how these processes are affected by microbial evolution. Here, we compared DNA sequences from snow/glacier algae found in an 8000-year-old ice from a glacier in central Asia with those from modern snow samples collected at 34 snow samples from globally distributed sites at the poles and mid-latitudes, to determine the evolutionary relationship between cosmopolitan and endemic phylotypes of snow algae. We further applied a coalescent theory-based demographic model to the DNA sequences. We found that the genus Raphidonema (Trebouxiophyceae) was distributed over both poles and mid-latitude regions and was detected in different ice core layers, corresponding to distinct time periods. Our results indicate that the modern cosmopolitan phylotypes belonging to Raphidonema were persistently present long before the last glacial period. Furthermore, endemic phylotypes originated from ancestral cosmopolitan phylotypes, suggesting that modern regional diversity of snow algae in the cryosphere is a product of microevolution. These findings suggest that the cosmopolitans dispersed across the world and then derived new localized endemics, which thus improves our understanding of microbial community formation by microevolution in natural environments.

Snow algae are photosynthetic microbes growing on melting snow surfaces, the blooms of which are visible on alpine snowpacks in Japan during the melting season. We characterized the seasonal and altitudinal variations in algal blooms on Mount Gassan, Japan, to assess the influence of vegetation on the algal bloom. From May to July in 2019, we collected colored snow from lower deciduous forest to the upper alpine areas. In the lower forest area, chlorophyll-a concentration in colored snow samples increased concomitantly with deciduous tree budburst, whereas in the alpine area, chlorophyll-a increased from June to July, although at lower levels than in the forest area. The absorption spectra of algal pigments extracted from samples differed among study sites and seasons, with those obtained from forest sites mainly characterized by absorption of chlorophylls and primary carotenoids, whereas those from alpine sites showed an intense secondary carotenoid peak throughout the study period. Chemical solute analyses revealed a general abundance of phosphate in snow, which was significantly higher in forest than in alpine sites and positively correlated with chlorophyll-a concentration. These findings suggest that nutrient supply from the forest canopy largely controls the appearance of colored snow in this area.

Abstract Snow ecosystems are an important component of polar and mountainous regions, influencing water regime, biogeochemical cycles and supporting snow specific taxa. Although snow is considered to be one of the most unique, and at the same time a disappearing habitat, knowledge of its taxonomic diversity is still limited. It is true especially for micrometazoans appearing in snow algae blooming areas. In this study, we used morphological and molecular approaches to identify two tardigrade species found in green snow patches of Mt. Gassan in Japan. By morphology, light (PCM) and scanning electron microscopy (SEM), and morphometry we described Hypsibiusnivalis sp. nov. which differs from other similar species by granular, polygonal sculpture on the dorsal cuticle and by the presence of cuticular bars next to the internal claws. Additionally, phylogenetic multilocus (COI, 18S rRNA, 28S rRNA) analysis of the second taxon, Hypsibius sp. identified by morphology as convergens-pallidus group, showed its affinity to the Hypsibiidae family and it is placed as a sister clade to all species in the Hypsibiinae subfamily. Our study shows that microinvertebrates associated with snow are poorly known and the assumption that snow might be inhabited by snow-requiring tardigrade taxa cannot be ruled out. Furthermore, our study contributes to the understanding subfamily Hypsibiinae showing that on its own the morphology of specimens belonging to convergens-pallidus group is insufficient in establishing a true systematic position of specimens.

Abstract Background Cryoconite granules are mineral–microbial aggregates found on glacier surfaces worldwide and are hotspots of biogeochemical reactions in glacier ecosystems. However, despite their importance within glacier ecosystems, the geographical diversity of taxonomic assemblages and metabolic potential of cryoconite communities around the globe remain unclear. In particular, the genomic content of cryoconite communities on Asia’s high mountain glaciers, which represent a substantial portion of Earth’s ice masses, has rarely been reported. Therefore, in this study, to elucidate the taxonomic and ecological diversities of cryoconite bacterial consortia on a global scale, we conducted shotgun metagenomic sequencing of cryoconite acquired from a range of geographical areas comprising Polar (Arctic and Antarctic) and Asian alpine regions. Results Our metagenomic data indicate that compositions of both bacterial taxa and functional genes are particularly distinctive for Asian cryoconite. Read abundance of the genes responsible for denitrification was significantly more abundant in Asian cryoconite than the Polar cryoconite, implying that denitrification is more enhanced in Asian glaciers. The taxonomic composition of Cyanobacteria, the key primary producers in cryoconite communities, also differs between the Polar and Asian samples. Analyses on the metagenome-assembled genomes and fluorescence emission spectra reveal that Asian cryoconite is dominated by multiple cyanobacterial lineages possessing phycoerythrin, a green light-harvesting component for photosynthesis. In contrast, Polar cryoconite is dominated by a single cyanobacterial species Phormidesmis priestleyi that does not possess phycoerythrin. These findings suggest that the assemblage of cryoconite bacterial communities respond to regional- or glacier-specific physicochemical conditions, such as the availability of nutrients (e.g., nitrate and dissolved organic carbon) and light (i.e., incident shortwave radiation). Conclusions Our genome-resolved metagenomics provides the first characterization of the taxonomic and metabolic diversities of cryoconite from contrasting geographical areas, highlighted by the distinct light-harvesting approaches of Cyanobacteria and nitrogen utilization between Polar and Asian cryoconite, and implies the existence of environmental controls on the assemblage of cryoconite communities. These findings deepen our understanding of the biodiversity and biogeochemical cycles of glacier ecosystems, which are susceptible to ongoing climate change and glacier decline, on a global scale.

Diverse microbes have been revealed to live in glaciers worldwide, but only a few biological studies were dedicated to glaciers in tropical Africa. These glaciers are shrinking rapidly and are expected to disappear shortly. In this study, we carried out biological and glaciological field observations on Stanley Glacier, the largest remaining glacier in the Rwenzori Mountains, Uganda, Africa. Microbial aggregates ranging from micrometer to centimeter in size were found on the glacier surface and contained moss and various types of Chlorophyta, among which a new endemic species of green alga. Concentrations of total impurities on the glacier surface, including microbial aggregates, varied spatially and decreased as altitude increased. The large microbial aggregates (larger than 4 cm in diameter) were found only at the glacier surface near the terminus and side margins, where the surface was less frequently covered with snow. It is also shown that the total organic matter on the glacier surface is determined by the timing of snow cover, which affects the quantity of solar radiation reaching the glacier ice surface. Furthermore, the total impurity content was negatively correlated with surface reflectivity, revealing their potential role in albedo reduction at the glacier surface through positive feedback between enhanced meltwater and increased biological growth.

Cryoconite is a mixture of mineral and organic material covering glacial ice, playing important roles in biogeochemical cycles and lowering the albedo of a glacier surface. Understanding the differences in structure of cryoconite across the globe can be important in recognizing past and future changes in supraglacial environments and ice-organisms-minerals interactions. Despite the worldwide distribution and over a century of studies, the basic characteristics of cryoconite, including its forms and geochemistry, remain poorly studied. The major purpose of our study is the presentation and description of morphological diversity, chemical and photoautotrophs composition, and organic matter content of cryoconite sampled from 33 polar and mountain glaciers around the globe. Observations revealed that cryoconite is represented by various morphologies including loose and granular forms. Granular cryoconite includes smooth, rounded, or irregularly shaped forms; with some having their surfaces covered by cyanobacteria filaments. The occurrence of granules increased with the organic matter content in cryoconite. Moreover, a major driver of cryoconite colouring was the concentration of organic matter and its interplay with minerals. The structure of cyanobacteria and algae communities in cryoconite differs between glaciers, but representatives of cyanobacteria families Pseudanabaenaceae and Phormidiaceae, and algae families Mesotaeniaceae and Ulotrichaceae were the most common. The most of detected cyanobacterial taxa are known to produce polymeric substances (EPS) that may cement granules. Organic matter content in cryoconite varied between glaciers, ranging from 1% to 38%. The geochemistry of all the investigated samples reflected local sediment sources, except of highly concentrated Pb and Hg in cryoconite collected from European glaciers near industrialized regions, corroborating cryoconite as element-specific collector and potential environmental indicator of anthropogenic activity. Our work supports a notion that cryoconite may be more than just simple sediment and instead exhibits complex structure with relevance for biodiversity and the functioning of glacial ecosystems. (c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

<title>Abstract</title>The results show the morphological analyses and spectroscopic studies of snow and glacier algae and their parasitic fungi in Svalbard (High Arctic). Fixed algal cells of two species, <italic>Sanguina nivaloides</italic> and <italic>Ancylonema nordenskioeldii,</italic> were imaged using light microscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM). Fluorescence microscopy using Calcofluor white stain supported the observations of parasitic fungi on the algal cells. Images in brightfield microscopy showed chytrid-like fungi penetrating the cells of both algal species. Parasites were found to colonize the cells of <italic>A. nordenskioeldii</italic> and hypnozygotes of <italic>S. nivaloides</italic>, while no fungi infected the cyst stages of <italic>S. nivaloides</italic>. The autofluorescence analysis revealed the ability of <italic>S. nivaloides</italic> to glow when excited with different wavelengths, while <italic>A. nordenskioeldii</italic> did not fluoresce. The hypnozygotes of <italic>S. nivaloides</italic> emitted brighter fluorescence than the cysts, and the most intense luminosity was observed in the UV range. The Fourier-transform infrared spectroscopy (FTIR) and energy-dispersive X-ray spectroscopy (EDS) spectroscopic analysis showed differences in the chemical composition between samples collected from three different sites. Samples dominated by cyst cells were characterized by the presence of an abundant polysaccharide envelope.

Previous reconstructions of summer temperatures from ice cores have relied on a statistical relationship between a melt layer and temperature observed at nearby stations. This study presents a novel method for reconstructing summer temperatures from melt layers in ice cores using an energy balance model that incorporates heat conduction and meltwater refreezing in the firn. We use the seasonal patterns in the ERA-Interim reanalysis data set for an ice core site to calculate the amounts of refreezing water within the firn under various summer mean temperature (SMT) and annual precipitation conditions, and prepared calibration tables containing these three variables. We then estimate the SMTs from the refreezing amount and annual accumulation, both of which can be obtained from an ice core. We apply this method to four ice cores that were recovered from sites with different climates: two sites on the Greenland Ice Sheet, one in Alaska, and one in Russian Altai. The reconstructed SMTs show comparable variations with those of observed temperatures at nearby stations. The nonlinear relationship between SMT and melt layer thickness differs between sites, indicating that a single linear approximation cannot be employed to estimate SMT. Sensitivity analyses suggest that the annual temperature range, amount of annual precipitation, and firn albedo (which is a time-invariant value in the model) significantly affect the relationship between SMT and melt layer thickness. This new method provides an alternative to existing approaches and yields an independent estimate of SMT from ice cores that have been affected by melting.

Although studies on snow algae and macroinvertebrates have been frequently conducted on snow patches, only few surveys have been focused on microinvertebrates which reach high biomass and play various trophic roles in other cold habitats. The aims of this study were (1) to search for microinvertebrates in seasonal surface snow patches located on the slope of Mt. Gassan, in northern Japan, and (2) to identify factors determining their distribution associated with snow algal blooms of various colorations (orange, green, and golden-brown) collected from the same sampling site over two seasons (2018, 2019). Microscopic observation revealed presence of two major groups of microinvertebrates: Tardigrada and Rotifera. They were concentrated in green snow colored by blooms of Chloromonas sp. in comparison to orange or golden-brown snow and only a few were found in white snow. Mean body length of tardigrades increased throughout the melt season, their intestine content was green and they laid eggs on colored snow. These results suggest that tardigrades preferentially grew and reproduced on green snow patches. Population densities of tardigrades, rotifers and concentration of chlorophyll a were significantly correlated. Our study indicates that green snow patches in temperate mountainous forests constitute important and unique low-temperature ecosystems for microinvertebrates. Snow covered by algae is an unrecognized novel habitats for tardigrades and rotifers.

Snow algae are photosynthetic microbes that inhabit the melting snow surface in alpine and polar regions. We analyzed the pigment and species composition of colored snow collected on Mt. Tateyama in Japan during the melting seasons of 2015 and 2016. High-performance liquid chromatographic analyses of the pigments extracted from the colored snow showed that their composition varied within the study area and were classified into four types: Type A (astaxanthin-monoester dominant), Type B (medium astaxanthin-monoester content), Type C (abundant primary carotenoids and free-astaxanthin), and Type D (abundant primary carotenoids and astaxanthin diesters). Types A and B were most commonly observed in the study area, whereas Types C and D appeared only at specific sites. Analysis of the 18S ribosomal RNA (18S rRNA) gene revealed six major amplicon sequence variants (ASVs) of snow algae, belonging to the Sanguina, Chloromonas, and Chlainomonas groups. The relative abundance of the algal ASVs showed that Sanguina was dominant (>48%) in both Types A and B, suggesting that the difference in astaxanthin abundance between the two types was caused by the production of pigments in the algal cells. The algal community structures of Types C and D differed from those of Types A and B, indicating that the primary carotenoids and astaxanthin diesters were derived from certain algal species in these types. Therefore, astaxanthin-rich Sanguina algae mostly induced the red snow that appeared widely in this alpine area; however, they were partially dominated by Chloromonas or Chlainomonas algae, causing different pigment compositions.

The worldwide distribution of microinvertebrates on glaciers, the coldest biome, is poorly known. Owing to their tolerance to hostile conditions, small size and dispersal abilities, nematodes, tardigrades and rotifers are considered cosmopolitan and together inhabit various ecosystems. In this study, we investigated their global distribution in cryoconite holes – a type of freshwater reservoir forming directly in the glacial ice that creates biodiversity hotspots on glaciers. We analysed cryoconite samples (using classical microscopic observations and environmental DNA metabarcoding) from 42 glaciers located around the world (the Arctic, Subarctic, Scandinavia, the Alps, the Caucasus, Siberia, Central Asia, Africa, South America and Antarctica), as well as using literature data. Samples from Antarctic, Karakoram and the Alps were analysed using next-generation sequencing (NGS) and classical observations under microscopes, while all other samples were analysed by microscope alone. Three general outcomes were found: (1) tardigrades and rotifers represented the most common invertebrates in cryoconite holes; (2) tardigrades and rotifers often coexisted together, with one or the other dominating, but the dominant taxon varied by region or by glacier; (3) nematodes – the most abundant, hyperdiverse and widespread metazoans on Earth, including in environments surrounding and seeding glacial surfaces – were consistently absent from cryoconite holes. Despite the general similarity of environmental conditions in cryoconite holes, the distribution of tardigrades and rotifers differed among glaciers, but not in any predictable way, suggesting that their distribution mostly depended on the random dispersal, extreme changes of supraglacial zone or competition. Although nematodes have been found in supraglacial habitats, cryoconite hole environments seem not to provide the necessary conditions for their growth and reproduction. Lack of physiological adaptations to permanently low temperatures (~0°C) and competition for different food resources in the cryoconite hole environment may explain the absence of nematodes in cryoconite holes.

Snow algae are photosynthetic microbes growing in thawing snow. They usually show various morphological cell types. The aim of this study was to carry out microscopic and spectroscopic analysis of different forms of cells of snow algae collected on glaciers in Alaska. Four different shapes of algal cells were observed with the use of bright field LM (Light Microscopy), DIC (Differential Interference Contrast), EDF (Extended Depth Focus), fluorescence microscopy, and SEM (Scanning Electron Microscopy). The cells exhibited the strongest autofluorescence after the exposure to 365-nm excitation light, and the intensity differed among the cell types. Zygotes (cysts) showed the most intense fluorescence. Acridine orange staining revealed the acid nature of the algal cells. The use of Congo red and Calcofluor white fluorochromes indicated differences in the structure of polysaccharides in the cell wall in the individual types of algal cells. FTIR (Fourier-Transform Infrared Spectroscopy) analyses showed the presence of polysaccharides not only in the algal cells but also in the fixative solution. The presence of polysaccharides in the extracellular algal fraction was confirmed by X-ray dispersion spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy imaging (SEM). The differences observed in the structure of the cell wall of the different forms of red snow algae prompt further analysis of this structure.

Cryoconite granules are naturally occurring microbial structures on glacier surfaces worldwide. They play a key role in carbon and nitrogen cycling in glacier ecosystems and can accelerate the melting of snow and ice. However, detailed mechanism of nitrogen cycling in cryoconite granules remains unclear. Here, we demonstrate that redox stratification affects the spatial distribution of N cycling processes in cryoconite granules. Based on microsensor measurements for O2, NH4 +, NO2 - and NO3 -, we identified the presence of fine-scale redox stratification within cryoconite granules. Cyanobacteria at the surface layer of the granules created oxic conditions, whereas the inner core of the granules was anoxic. Metatranscriptomic analyses indicated the active occurrences of nitrification in the inner core, whereas denitrification actively occurred both in the inner core and the surface layer of the granules. Cyanobacteria in the inner core of the granules were inactive, and likely dead and being degraded, providing carbon and nitrogen to support nitrifiers and denitrifiers. Quantities of nitrification genes/transcripts were greater in large cryoconite granules than small ones, most likely because nitrogen substrates were more abundantly present in the inner core of large granules due to distinct redox stratification. Our results suggest that the development of a granular structure of cryoconite granules can largely affect carbon and nitrogen cycling on glaciers.

To understand the microbial composition and diversity patterns, cryoconite granules were collected from two geographical areas, i.e., Nepali Himalaya and Greenland, Arctic. 16S rRNA, ITS and the D1/D2 domain sequencing techniques were used for characterization of microbial communities of the four glaciers. The total 13 species of bacteria such as Bacillus aryabhattai, Bacillus simplex, Brevundimonas vesicularis, Cryobacterium luteum, Cryobacterium psychrotolerans, Dermacoccus nishinomiyaensis, Glaciihabitans tibetensis, Leifsonia kafniensis, Paracoccus limosus, Polaromonas glacialis, Sporosarcina globispora, Staphylococcus saprophyticus, Variovorax ginsengisoli, and 4 species of fungi such as Goffeauzyma gilvescens, Mrakia robertii, Dothideomycetes sp., Helotiales sp. were recorded from Nepali Himalaya. Among these, 12 species of bacteria and 4 species of fungi are new contributions to Himalaya. In contrast to this, six species of bacteria such as Bacillus cereus, Cryobacterium psychrotolerans, Dermacoccus nishinomiyaensis, Enhydrobacter aerosaccus, Glaciihabitans tibetensis, Subtercola frigoramans, and nine species of fungi such as Goffeauzyma gilvescens, Mrakia robertii, Naganishia vaughanmartiniae, Piskurozyma fildesensis, Rhodotorula svalbardensis, Alatospora acuminata, Articulospora sp., Phialophora sp., Thelebolus microspores, and Dothideomycetes sp.), were recorded from Qaanaaq, Isunnguata Sermia and Thule glaciers, Greenland. Among these, five species of bacteria and seven species of fungi are new contributions to Greenland cryoconite. Microbial analyses indicate that the Nepali Himalayan cryoconite colonize higher numbers of microbial species compared to the Greenland cryoconite.

Cryoconite, a sediment found on the surface of glaciers, is known for its ability to accumulate radionuclides. New data on cryoconite from the Morteratsch glacier (Switzerland) are presented to shed light on the mechanisms that control the distribution of radioactivity in cryoconite. Among the radionuclides detected in our samples, we have identified Ag, an artificial species which has never been observed in terrestrial environments before. This finding supports that cryoconite has an extraordinary ability to accumulate radioactivity. Our results also show that the radioactivity of cryoconite from a single glacier is far from uniform. Both the absolute amount of radioactivity and the relative contribution of single radionuclides are highly variable in samples from the Morteratsch glacier. To investigate the processes responsible for such variability, we have explored the correlation between radionuclides, organic and inorganic carbon fractions and the morphological features of cryoconite deposits. We have found that the degree of connection between cryoconite and supraglacial hydrology is particularly important, since it strongly influences the accumulation of radionuclides in cryoconite. Cryoconite holes connected with supraglacial channels are rich in cosmogenic Be; in contrast, poorly connected deposits are rich in artificial fallout radionuclides and elemental carbon. The very different half-lives of Be and artificial radionuclides allowed us to discuss our findings in relation to the age and maturity of cryoconite deposits, highlighting the potential use of radionuclides to investigate hydrological supraglacial processes and material cycling at the surface of glaciers. 108m 7 7

Surface albedo of snow and ice is substantially reduced by inorganic impurities, such as aeolian mineral dust (MD) and black carbon (BC), and also by organic impurities, such as microbes that live in the snow. In this paper, we present the temporal changes of surface albedo, snow grain size, MD, BC and snow algal cell concentration observed on a snowpack in northwest Greenland during the ablation season of 2014 and our attempt to reproduce the changes in albedo with a physically based snow albedo model. We also attempt to reproduce the effects of inorganic impurities and the red snow algae (Sanguina nivaloides) on albedo. Concentrations of MD and red snow algae in the surface snow were found to increase in early August, while snow grain size and BC were found to not significantly change throughout the ablation season. Surface albedo was found to have decreased by 0.08 from late July to early August. The albedo simulated by the model agreed with the albedo observed during the study period. However, red snow algae exerted little effect on surface albedo in early August. This is probably owing to the abundance of smaller cells (<span classCombining double low line"inline-formula">4.9×104</span> cells L<span classCombining double low line"inline-formula">-1</span>) when compared with the cell abundance of red snow reported by previous studies in the Arctic region (<span classCombining double low line"inline-formula">1/4108</span> cells L<span classCombining double low line"inline-formula">-1</span>). The simulation of snow albedo until the end of the melting season, with a snow algae model, revealed that the reduction in albedo attributed to red snow algae could equal 0.004, out of a total reduction of 0.102 arising from the three impurities on a snowpack in northwest Greenland. Finally, we conducted scenario simulations using the snow albedo model, coupled with the snow algae model, in order to simulate the possible effects of red snow blooming on snow albedo under warm conditions in northwest Greenland. The result suggests that albedo reduction by red snow algal growth under warm conditions (surface snow temperature of <span classCombining double low line"inline-formula">+</span>1.5 <span classCombining double low line"inline-formula"></span>C) reached 0.04, equivalent to a radiative forcing of 7.5 W m<span classCombining double low line"inline-formula">-2</span> during the ablation season of 2014. This coupled albedo model has the potential to dynamically simulate snow albedo, including the effect of organic and inorganic impurities, leading to proper estimates of the surface albedo of snow cover in Greenland.

Surface albedo is an important driver of surface processes that promote glacier melting and is a key variable influencing glacier surface melt. Despite much focus in the literature on variations in albedo and its influence on snow surfaces, little attention has been paid to dust and its impact on bare-ice albedo with respect to glacier melting surfaces. In this paper, spatial changes in glacier albedo were investigated using three Landsat images taken during the ablation season in 2006; temporal variations in albedo were measured by an automatic weather station (AWS) in the ablation zone between 26 June and 1 August 2007 at Urumqi Glacier No. 1 in Tien Shan. Ice and snow samples and reflection spectra at 325–1050 nm were collected in August, 2007 at Urumqi Glacier No. 1. The data suggested that spatial changes in glacier albedo are not prominent after snowfall; however, once ice becomes exposed, glacier albedo varies remarkably and generally increases with elevation, especially around the snow line. Temporal variations are characterized by a large range and high frequency, and most are induced by snowfall, changes in cloud conditions, and surface dust; snowfall and cloud increase glacier albedo. Furthermore, the response of snow albedo is more sensitive to cloud compared with the response of ice albedo. Over a bare ice surface, the albedo generally decreases as the concentration of surface dust increases. Organic matter is a primary factor in reducing the albedo over ice.

The physical and geochemical characteristics of shallow ice cores drilled in the accumulation area (Western Cwm) of Khumbu Glacier in the Nepal Himalayas in 1980 were analyzed. The two ice cores from different elevations (cores 1 and 2 from 6,100 and 6,400 m a.s.l., respectively) showed distinct stratigraphy: Core 1 consisted of stratified firn layers with a few ice and dust layers, whereas core 2 consisted of unstratified refrozen ice with abundant debris. The mean oxygen-stable isotope was significantly lower in core 2 than in core 1, and the composition of major soluble ions also differed between the ice cores. The mineralogical characteristics of debris in core 2 were in accordance with the geology of the south face of Mt. Everest, indicating that it was supplied from above 7,000 m a.s.l. by avalanches. The distinct stratigraphy and geochemistry suggest that the snow accumulation process differed between the two drilling sites; snow was supplied by precipitation in core 1, whereas in core 2 it was supplied by avalanche with rock debris from the higher site of the south face of Mt. Everest. The two distinct snow accumulation processes are likely to characterize the mass balance and formation of debris covers of the lower ablation area of Khumbu Glacier.

Abstract. Surface albedo of snow and ice is substantially reduced by inorganic impurities, such as aeolian mineral dust (MD) and black carbon (BC), and also by organic impurities, such as microbes that live in the snow. In this paper, we present the temporal changes of surface albedo, snow grain size, MD, BC, and snow algal cell concentration observed on a snowpack in northwest Greenland during the ablation season of 2014 and our attempt to reproduce the changes in albedo with a physically based snow albedo model. We also attempt to reproduce the effects of inorganic impurities and the red snow algae (Chlamydomonas nivalis) on albedo. Concentrations of MD and red snow algae in the surface snow were found to increase in early August, while snow grain size and BC were found to not significantly change throughout the ablation season. Surface albedo was found to have decreased by 0.08 from late July to early August. The albedo simulated by the model agreed with the albedo observed during the study period. However, red snow algae exerted little effect on surface albedo in early August. This is probably owing to the abundance of smaller cells (4.9 × 104 cells L^-1) when compared with the cell abundance of typical red algal snow (~ 108 cells L−1). The simulation of snow albedo until the end of the melting season, with an algal growth model, revealed that the reduction in albedo attribute to red algae could equal 0.004, out of a total reduction of 0.102 arising from the three impurities on a snowpack in northwest Greenland. Finally, we conducted scenario simulations using the snow albedo model, coupled with the algal growth model, in order to simulate the possible effects of red algal blooming on snow albedo under warm conditions in northwest Greenland. The result suggests that albedo reduction by red snow algal growth under warm conditions (surface snow temperature of +1.5 °C) reached 0.04, equivalent to a radiative forcing of 7.5 W m−2 during the ablation season of 2014. This coupled albedo model has the potential to dynamically simulate snow albedo, including the effect of organic and inorganic impurities, leading to proper estimates of the surface albedo of snow cover in Greenland.

Cryoconite granules are dark-colored biological aggregates on glaciers. Bacterial community varies with granule size, however, community change in space and their susceptibility to environmental factors has not been described yet. Therefore, we focused on bacterial community from four different granule sizes (30-249 μm, 250-750 μm, 750-1599 μm, more than 1600 μm diameter) in 10 glaciers in northwestern Greenland and their susceptibility to exogenous nutrients in cryoconite hole. A filamentous cyanobacterium Phormidesmis priestleyi, which has been frequently reported from glaciers in Arctic was abundant (10%-26%) across any size of granules on most of glaciers. Bacterial community across glaciers became similar with size increase, and whence smallest size fractions contain more unique genera in each glacier. Multivariate analysis revealed that effect of nutrients to beta diversity is larger in smaller granules (30-249 μm and 250-750 μm diameter), suggesting that bacterial susceptibility to nutrients changes with growth of granule (i.e. P. priestleyi was affected by nitrate in early growth stage).

Cryoconite granules, which are dark-colored biological aggregates on glaciers, effectively accelerate the melting of glacier ice. Bacterial community varies with granule size, however, community change in space and their susceptibility to environmental factors has not been described yet. Therefore, we focused on bacterial community from 4 different granule sizes (30-249 μm, 250-750 μm, 750-1599 μm, more than 1600 μm diameter) in 10 glaciers in northwestern Greenland and their susceptibility for exogenous nutrients in cryoconite hole. A filamentous cyanobacterium Phormidesmis priestleyi, which has been frequently reported from glaciers in Arctic was abundant (10-26%) across any size of granules on most of glaciers. Bacterial community across glaciers became similar with size increase, and whence smallest size fractions contain more unique genera in each glacier. Multivariate analysis suggests that phosphate, which is significantly higher in one glacier (Scarlet Heart Glacier), is primary associated with bacterial beta diversity. Correlation coefficients between abundance of major genera and nutrients largely changed with granule size, suggesting that nutrients susceptibility to genera changes with growth process of granule (e.g. P. priestleyi was affected by nitrate in early growth stage).

Pollen grains are commonly found in ice cores, particularly those from mountain glaciers at low to middle latitudes. Because the release of pollen from flowers has a seasonality and varies among the plant species, pollen concentrations in ice cores are useful for distinguishing annual or seasonal layers and inferring past vegetation near glaciers. We analyzed major pollen grains in an 87-m-deep ice core drilled at the top of the Grigoriev Ice Cap (4563 m.a.s.l.) in the Tien Shan Mountains, Kyrgyz Republic. Microscopy showed that mainly five pollen taxa were contained in the ice core. Their abundance fluctuated with the depth of the core, indicating their seasonal deposition on the ice cap, while the seasonality of the stable isotopes was not particularly clear because of melting and refreezing signatures. Based on the pollen profiles, annual layers were determined back in time to 1780 AD at a depth of 63.6 m; at greater depth, they could not be distinguished due to ice layer thinning in the glacier. The pollen assemblages have gradually changed during the last 220 years and were particularly distinctive in the deeper part, suggesting the drastic change of vegetation in this region during the Holocene.

Red-snow algae are red-pigmented unicellular algae that appear seasonally on the surface of thawing snow worldwide. Here, we analyse the distribution patterns of snow algae sampled from glaciers and snow patches in the Arctic and Antarctica based on nuclear ITS2 sequences, which evolve rapidly. The number of phylotypes is limited in both polar regions, and most are specific to either the Arctic or Antarctica. However, the bipolar phylotypes account for the largest share (37.3%) of all sequences, suggesting that red-algal blooms in polar regions may comprise mainly cosmopolitan phylotypes but also include endemic organisms, which are distributed either in the Arctic or Antarctica.

The glacier stonefly Andiperla willinki is the largest metazoan inhabiting the Patagonian glaciers. In this study, we analysed the gut microbiome of the aquatic nymphs by 16S rRNA gene amplicon and metagenomic sequencing. The bacterial gut community was consistently dominated by taxa typical of animal digestive tracts, such as Dysgonomonadaceae and Lachnospiraceae, as well as those generally indigenous to glacier environments, such as Polaromonas. Interestingly, the dominant Polaromonas phylotypes detected in the stonefly gut were almost never detected in the glacier surface habitat. Fluorescence in situ hybridization analysis revealed that the bacterial lineages typical of animal guts colonized the gut wall in a co-aggregated form, while Polaromonas cells were not included in the aggregates. Draft genomes of several dominant bacterial lineages were reconstructed from metagenomic datasets and indicated that the predominant Dysgonomonadaceae bacterium is capable of degrading various polysaccharides derived from host-ingested food, such as algae, and that other dominant bacterial lineages ferment saccharides liberated by the polysaccharide degradation. Our results suggest that the gut bacteria–host association in the glacier stonefly contributes to host nutrition as well as material cycles in the glacier environment.

To understand the geological origins of minerals in cryoconite and the nutrients sources for microbes on glaciers, we analyzed the Sr-Nd isotopic ratios of the four mineral fractions in cryoconites including saline, carbonate, phosphate, silicate and the organic fraction obtained from Gulkana Glacier in Alaska. The isotopes in the silicate mineral fraction exhibited spatial variation within the glacier ( Sr/ Sr: 0.704533-0.709563, ÎNd (0): -16.0 to 0.5), which can be explained by the different mixing ratios of the two distinct sources: one of the sources is lateral and terminal moraines or soil, and the other is the medial moraine of the glacier. The minerals in the cryoconite at the lower sites in the glacier are likely derived from the former source, whereas those at the upper sites are from latter sources. The mineralogical and elemental compositions also support mixing of the silicate minerals from the two local sources. The Sr isotopic ratios of the organic fraction also showed spatial variation on the glacier in the middle sites - a trend similar to those of the phosphate fraction. The results suggest that the organic matter is mostly the byproducts of microbes using the phosphate minerals as a nutrient source. 87 86

Cryoconite holes are water-filled cylindrical holes formed on ablation ice surfaces and commonly observed on glaciers worldwide. Temporal changes of cryoconite holes characteristically <5 cm in diameter were monitored with a time-lapse interval camera over 15 d during the melting season on Qaanaaq Glacier in northwest Greenland. The holes drastically changed their dimensions and synchronously collapsed twice during the study period. When the holes collapsed, the coverage of cryoconite on the ice surface increased from 1.0 to 3.5% in the field of view of the camera, and then decreased again to 0.4% after the holes reformed. Comparison with meteorological data showed that the collapses occurred in cloudy and rainy or windy weather conditions, corresponding to low shortwave solar radiation (68-126 W m , 40-55% of the incoming flux). In contrast, holes developed in sunny conditions correspond to high solar radiation (186-278 W m , 63-88%). Results suggest that the dimensions of holes drastically changed depending on the weather conditions and that frequent cloudy, warm and windy conditions would cause a decay of holes and weathering crust, inducing an increase in the cryoconite coverage on the ice, consequently darkening the glacier surface. -2 -2

Abstract. Snow algal bloom is a common phenomenon on melting snowpacks in polar and alpine regions and can substantially increase snow melt rates due to the effect of albedo reduction on the snow surface. In order to reproduce algal growth on the snow surface using a numerical model, temporal changes in snow algal abundance were investigated on the Qaanaaq Glacier in north-western Greenland from June to August 2014. Snow algae first appeared at the study sites in late June, which was approximately 94 h after air temperatures exceeded the melting point. Algal abundance increased exponentially after this appearance, but the increasing rate became slow after late July, and finally reached 3.5 × 107 cells m−2 in early August. We applied a logistic model to the algal growth curve and found that the algae could be reproduced with an initial cell concentration of 6.9 × 102 cells m−2, a growth rate of 0.42 d−1, and a carrying capacity of 3.5 × 107 cells m−2 on this glacier. This model has the potential to simulate algal blooms from meteorological data sets and to evaluate their impact on the melting of seasonal snowpacks and glaciers.

Despite the crucial role of cyanobacteria in various ecosystems, little is known about their evolutionary histories, especially microevolutionary dynamics, because of the lack of knowledge regarding their mutation rates. Here we directly estimated cyanobacterial mutation rates based on ancient DNA analyses of ice core samples collected from Kyrgyz Republic that dates back to ~12,500 cal years before present. We successfully sequenced the 16S rRNA and 16S-23S internal transcribed spacer (ITS) region. Two cyanobacterial operational taxonomic units (OTUs) were detected from the ancient ice core samples, and these OTUs are shared with those from the modern glacier surface. The mutation rate of ITS region was estimated by comparing ancient and modern populations, and were at the magnitude of 10 substitutions/sites/year. By using a model selection framework, we also demonstrated that the ancient sequences from the ice sample were not contaminated from modern samples. Bayesian demographic analysis based on coalescent theory revealed that cyanobacterial population sizes increased over Asia regions during the Holocene. Thus, our results enhance our understanding of the enigmatic timescale of cyanobacterial microevolution, which has the potential to elucidate the environmental responses of cyanobacteria to the drastic climatic change events of the Quaternary. -7

Intensive groundwater development in the urban area of the Nagaoka Plain, Japan, has induced changes in the pH and saturation index of calcite in groundwater. To account for these chemical changes, it is important to determine seasonal variations of recharge and the groundwater flow system in the aquifer. This study identified the sources and flow system of groundwater in this urban area by a comprehensive method using stable isotope data and a numerical groundwater model of the Nagaoka Plain. Stable isotope evidence shows that the groundwater is recharged by meteoric water originating from low-elevation areas rather than the mountains surrounding the plain. The water table in the study area is drawn down during the winter and recovers in the other seasons. Numerical modeling shows that discharge occurs primarily along the Shinano River during the recovery period, whereas discharge is centered in urbanized areas during the drawdown period, when a conical depression of the water table stimulates recharge from the immediate area. These results are indications of a local groundwater flow system, with its recharge area between the Shinano River and the urban areas, which is governed by intensive seasonal groundwater extraction.