Potential sources of bacteria colonizing the cryoconite of an Alpine glacier.

We investigated the potential contribution of ice-marginal environments to the microbial communities of cryoconite holes, small depressions filled with meltwater that form on the surface of Forni Glacier (Italian Alps). Cryoconite holes are considered the most biologically active environments on glaciers. Bacteria can colonize these environments by short-range transport from ice-marginal environments or by long-range transport from distant areas. We used high throughput DNA sequencing to identify Operational Taxonomic Units (OTUs) present in cryoconite holes and three ice-marginal environments, the moraines, the glacier forefield, and a large (> 3 m high) ice-cored dirt cone occurring on the glacier surface. Bacterial communities of cryoconite holes were different from those of ice-marginal environments and hosted fewer OTUs. However, a network analysis revealed that the cryoconite holes shared more OTUs with the moraines and the dirt cone than with the glacier forefield. Ice-marginal environments may therefore act as sources of bacteria for cryoconite holes, but differences in environmental conditions limit the number of bacterial strains that may survive in them. At the same time, cryoconite holes host a few OTUs that were not found in any ice-marginal environment we sampled, thus suggesting that some bacterial populations are positively selected by the specific environmental conditions of the cryoconite holes.

Temporal variability of bacterial communities in cryoconite on an alpine glacier.

Cryoconite holes, that is, small ponds that form on glacier surface, are considered the most biologically active environments on glaciers. Bacterial communities in these environments have been extensively studied, but often through snapshot studies based on the assumption of a general stability of community structure. In this study, the temporal variation of bacterial communities in cryoconite holes on the Forni Glacier (Italian Alps) was investigated by high throughput DNA sequencing. A temporal change of bacterial communities was observed with autotrophic Cyanobacteria populations dominating communities after snowmelt, and heterotrophic Sphingobacteriales populations increasing in abundance later in the season. Bacterial communities also varied according to hole depth and area, amount of organic matter in the cryoconite and oxygen concentration. However, variation in environmental features explained a lower fraction of the variation in bacterial communities than temporal variation. Temporal change along ablation season seems therefore more important than local environmental conditions in shaping bacterial communities of cryoconite of the Forni Glacier. These findings challenge the assumption that bacterial communities of cryoconite holes are stable.

Diversity and Assembling Processes of Bacterial Communities in Cryoconite Holes of a Karakoram Glacier.

Cryoconite holes are small ponds that form on the surface of glaciers that contain a dark debris, the cryoconite, at the bottom and host active ecological communities. Differences in the structure of bacterial communities have been documented among Arctic and mountain glaciers, and among glaciers in different areas of the world. In this study, we investigated the structure of bacterial communities of cryoconite holes of Baltoro Glacier, a large (62 km in length and 524 km2 of surface) glacier of the Karakoram, by high-throughput sequencing of the V5-V6 hypervariable regions of the 16S rRNA gene. We found that Betaproteobacteria dominated bacterial communities, with large abundance of genera Polaromonas, probably thanks to its highly versatile metabolism, and Limnohabitans, which may have been favoured by the presence of supraglacial lakes in the area where cryoconite holes were sampled. Variation in bacterial communities among different sampling areas of the glacier could be explained by divergent selective processes driven by variation in environmental conditions, particularly pH, which was the only environmental variable that significantly affected the structure of bacterial communities. This variability may be due to both temporal and spatial patterns of variation in environmental conditions.

Future hydrological regimes and glacier cover in the Everest region: The case study of the upper Dudh Koshi basin.

Assessment of future water resources under climate change is required in the Himalayas, where hydrological cycle is poorly studied and little understood. This study focuses on the upper Dudh Koshi river of Nepal (151km(2), 4200-8848ma.s.l.) at the toe of Mt. Everest, nesting the debris covered Khumbu, and Khangri Nup glaciers (62km(2)). New data gathered during three years of field campaigns (2012-2014) were used to set up a glacio-hydrological model describing stream flows, snow and ice melt, ice cover thickness and glaciers' flow dynamics. The model was validated, and used to assess changes of the hydrological cycle until 2100. Climate projections are used from three Global Climate Models used in the recent IPCC AR5 under RCP2.6, RCP4.5 and RCP8.5. Flow statistics are estimated for two reference decades 2045-2054, and 2090-2099, and compared against control run CR, 2012-2014. During CR we found a contribution of ice melt to stream flows of 55% yearly, with snow melt contributing for 19%. Future flows are predicted to increase in monsoon season, but to decrease yearly (-4% vs CR on average) at 2045-2054. At the end of century large reduction would occur in all seasons, i.e. -26% vs CR on average at 2090-2099. At half century yearly contribution of ice melt would be on average 45%, and snow melt 28%. At the end of century ice melt would be 31%, and snow contribution 39%. Glaciers in the area are projected to thin largely up to 6500ma.s.l. until 2100, reducing their volume by -50% or more, and their ice covered area by -30% or more. According to our results, in the future water resources in the upper Dudh Koshi would decrease, and depend largely upon snow melt and rainfall, so that adaptation measures to modified water availability will be required.

Light-dependent microbial metabolisms drive carbon fluxes on glacier surfaces.

Biological processes on glacier surfaces affect glacier reflectance, influence surface energy budget and glacier response to climate warming, and determine glacier carbon exchange with the atmosphere. Currently, carbon balance of supraglacial environment is assessed as the balance between the activity of oxygenic phototrophs and the respiration rate of heterotrophic organisms. Here we present a metagenomic analysis of tiny wind-blown supraglacial sediment (cryoconite) from Baltoro (Pakistani Karakoram) and Forni (Italian Alps) glaciers, providing evidence for the occurrence in these environments of different and previously neglected metabolic pathways. Indeed, we observed high abundance of heterotrophic anoxygenic phototrophs, suggesting that light might directly supplement the energy demand of some bacterial strains allowing them to use as carbon source organic molecules, which otherwise would be respired. Furthermore, data suggest that CO2 could be produced also by microbiologically mediated oxidation of CO, which may be produced by photodegradation of organic matter.

Influence of abiotic variables on culturable yeast diversity in two distinct Alpine glaciers.

The influence of some abiotic variables (pH, dry weight, organic carbon, nitrogen and phosphorous) on culturable yeast diversity in two distinct, but adjacent Alpine glaciers (Glacier du Géant, France, and Miage Glacier, Italy) was investigated. In all, 682 yeast strains were isolated and identified by D1/D2 and ITS sequencing as belonging to species of the genera Aureobasidium, Candida, Bulleromyces, Cryptococcus, Cystofilobasidium, Dioszegia, Guehomyces, Holtermanniella, Leucosporidiella, Mrakia, Mrakiella, Rhodotorula, Sporidiobolus, Sporobolomyces and Udenyomyces. Overall, the most represented genera were Cryptococcus (55% of isolates), Rhodotorula (17%) and Mrakia (10%). About 10% of strains, presumably belonging to new species (yet to be described), were preliminarily identified at the genus level. Principal component analysis (PCA) revealed that organic carbon, nitrogen and phosphorous are apparently mostly related to culturable yeast abundance and diversity. In this context, the hypothesis that the frequency of isolation of certain species may be correlated with some organic nutrients (with special emphasis for phosphorous) is discussed.

Bacterial community structure on two alpine debris-covered glaciers and biogeography of Polaromonas phylotypes.

High-elevation cold environments are considered ideal places to test hypotheses about mechanisms of bacterial colonization and succession, and about bacterial biogeography. Debris-covered glaciers (glaciers whose ablation area is mainly covered by a continuous layer of rock debris fallen from the surrounding mountains) have never been investigated in this respect so far. We used the Illumina technology to analyse the V5 and V6 hypervariable regions of the bacterial 16S rRNA gene amplified from 38 samples collected in July and September 2009 at different distances from the terminus on two debris-covered glaciers (Miage and Belvedere--Italian Alps). Heterotrophic taxa-dominated communities and bacterial community structure changed according to ice ablation rate, organic carbon content of the debris and distance from the glacier terminus. Bacterial communities therefore change during downwards debris transport, and organic carbon of these recently exposed substrates is probably provided more by allochthonous deposition of organic matter than by primary production by autotrophic organisms. We also investigated whether phylotypes of the genus Polaromonas, which is ubiquitous in cold environments, do present a biogeographical distribution by analysing the sequences retrieved in this study together with others available in the literature. We found that the genetic distance among phylotypes increased with geographic distance; however, more focused analyses using discrete distance classes revealed that both sequences collected at sites <100 km and at sites 9400-13,500 km to each other were more similar than those collected at other distance classes. Evidences of biogeographic distribution of Polaromonas phylotypes were therefore contrasting.

Assessment and selection of geomorphosites and trails in the Miage Glacier area (Western Italian alps).

Glacial environments are considered geomorphosites because they exhibit all of the features that characterise sites of geomorphological interest. The Miage Glacier, in particular, is the most important debris-covered glacier of the Italian Alps, and it has been extensively studied since the 18th century because of its scientific features. In this area, the geomorphological and geological attributes are evaluated at 11 sites that have been individuated along the three main touristic trails, which allow an exploration of the surroundings of the glacial tongue and its two main lobes. Using a methodology previously tested in a fluvial environment, single sites and trails are quantitatively assessed to determine the most suitable trail for educational purposes. Hazards that could potentially affect the trails are considered in terms of both risk education and final selection of the most suitable trails for the various possible user groups. The richness of scientific data in this area should increase its importance as a geomorphosite by increasing the educational value of the Miage Glacier and the Veny Valley.

Evaluating tourist perception of environmental changes as a contribution to managing natural resources in glacierized areas: a case study of the Forni glacier (Stelvio National Park, Italian Alps).

Climate change effects are noticeably evident above the timberline where glacier and permafrost processes and mass movements drive the surface evolution. In particular, the cryosphere shrinkage is deeply changing the features and characteristics of several glacierized mountain areas of the world, and these modifications can also affect the landscape perception of tourists and mountaineers. On the one hand glacier retreat is increasing the interest of tourists and visitors in areas witnessing clear climate change impacts; on the other hand cryosphere shrinkage can impact the touristic appeal of mountain territories which, diminishing their ice and snow coverage, are also losing part of their aesthetic value. Then, to promote glacierized areas in a changing climate and to prepare exhaustive and actual proposals for sustainable tourism, it is important to deepen our knowledge about landscape perception of tourists and mountaineers and their awareness of the ongoing environmental modifications. Here we present the results from a pilot study we performed in summer 2009 on a representative glacierized area of the Alps, the Forni Valley (Stelvio National Park, Lombardy, Italy), a valley shaped by Forni, the largest Italian valley glacier. During the 2009 summer season we asked tourists visiting the Forni Valley to complete a questionnaire. This study was aimed at both describing the features and characteristics of tourists and mountaineers visiting this Alpine zone in summer and evaluating their landscape perception and their ability to recognize climate change impacts and evidence. Our results suggest that the dissemination strategies in a natural protected area have to take into account not only the main landscape features but also the sites where the information will be given. In particular considering the peculiarities of the huts located in the area, such as their different accessibility and the fact that they are included or not in a mountaineering network like that of the Italian Alpine Club. Both these factors can influence the kind of visitors to the area, thus requiring different dissemination strategies. Moreover, differences in the viewpoints from where visitors could watch and understand landscape also have to be considered. Next, in a protected area where climate change effects are evident, the dissemination strategies should be developed in close cooperation with scientists who are analyzing the area and with the support of periodic interviews which could be very useful to evaluate the effectiveness of the applied dissemination methods. Last but not least, the questionnaire should be standardized and distributed in several protected areas, thus permitting useful comparisons and the identification of common solutions for sharing in a friendly way scientific knowledge about climate change and its effects on the environment and the landscape.

Yeast and yeast-like diversity in the southernmost glacier of Europe (Calderone Glacier, Apennines, Italy).

The present study reports the characterization of psychrophilic yeast and yeast-like diversity in cold habitats (superficial and deep sediments, ice cores and meltwaters) of the Calderone Glacier (Italy), which is the southernmost glacier in Europe. After incubation at 4 and 20 degrees C, sediments contained about 10(2)-10(3) CFU of yeasts g(-1). The number of viable yeast cells in ice and meltwaters was several orders of magnitude lower. The concomitant presence of viable bacteria and filamentous fungi has also been observed. In all, 257 yeast strains were isolated and identified by 26S rRNA gene D1/D2 and internal transcribed spacers (1 and 2) sequencing as belonging to 28 ascomycetous and basidiomycetous species of 11 genera (Candida, Cystofilobasidium, Cryptococcus, Dioszegia, Erythrobasidium, Guehomyces, Mastigobasidium, Mrakia, Mrakiella, Rhodotorula and Sporobolomyces). Among them, the species Cryptococcus gastricus accounted for almost 40% of the total isolates. In addition, 12 strains were identified as belonging to the yeast-like species Aureobasidium pullulans and Exophiala dermatitidis, whereas 15 strains, presumably belonging to new species, yet to be described, were also isolated. Results herein reported indicate that the Calderone Glacier, although currently considered a vanishing ice body due to the ongoing global-warming phenomenon, still harbors viable psychrophilic yeast populations. Differences of yeast and yeast-like diversity between the glacier under study and other worldwide cold habitats are also discussed.

Accelerating climate change impacts on alpine glacier forefield ecosystems in the European Alps.

In the European Alps the increase in air temperature was more than twice the increase in global mean temperature over the last 50 years. The abiotic (glacial) and the biotic components (plants and vegetation) of the mountain environment are showing ample evidence of climate change impacts. In the Alps most small glaciers (80% of total glacial coverage and an important contribution to water resources) could disappear in the next decades. Recently climate change was demonstrated to affect higher levels of ecological systems, with vegetation exhibiting surface area changes, indicating that alpine and nival vegetation may be able to respond in a fast and flexible way in response to 1-2 degrees C warming. We analyzed the glacier evolution (terminus fluctuations, mass balances, surface area variations), local climate, and vegetation succession on the forefield of Sforzellina Glacier (Upper Valtellina, central Italian Alps) over the past three decades. We aimed to quantify the impacts of climate change on coupled biotic and abiotic components of high alpine ecosystems, to verify if an acceleration was occurring on them during the last decade (i.e., 1996-2006) and to assess whether new specific strategies were adopted for plant colonization and development. All the glaciological data indicate that a glacial retreat and shrinkage occurred and was much stronger after 2002 than during the last 35 years. Vegetation started to colonize surfaces deglaciated for only one year, with a rate at least four times greater than that reported in the literature for the establishment of scattered individuals and about two times greater for the well-established discontinuous early-successional community. The colonization strategy changed: the first colonizers are early-successional, scree slopes, and perennial clonal species with high phenotypic plasticity rather than pioneer and snowbed species. This impressive acceleration coincided with only slight local summer warming (approximately -0.5 degree C) and a poorly documented local decrease in the snow cover depth and duration. Are we facing accelerated ecological responses to climatic changes and/or did we go beyond a threshold over which major ecosystem changes may occur in response to even minor climatic variations?

Psychrophilic yeasts in glacial environments of Alpine glaciers.

The presence of psychrophilic yeasts in supra- and subglacial sediments, ice and meltwater collected from two glaciers of the Italian Alps (Forni and Sforzellina-Ortles-Cevedale group) was investigated. After incubation at 4 degrees C, subglacial sediments contained from 1.3 x 10(3) to 9.6 x 10(3) CFU of yeasts g(-1). The number of yeast cells in supraglacial sediments was c. 10-100-fold lower. A significant proportion of isolated yeasts exhibited one or more extracellular enzymatic activities (starch-degrading, lipolytic, esterolytic, proteolytic and pectinolytic activity) at 4 degrees C. Selected isolates were able to grow at 2 degrees C under laboratory-simulated in situ conditions. In all, 106 isolated yeasts were identified by MSP-PCR fingerprinting and 26S rRNA gene sequencing of the D1/D2 region as belonging to 10 species: Aureobasidium pullulans, Cryptococcus gilvescens (over 50% of the total), Cryptococcus terricolus, Mrakia gelida, Naganishia globosa, Rhodotorula glacialis, Rhodotorula psychrophenolica, Rhodotorula bacarum, Rhodotorula creatinivora and Rhodotorula laryngis. Four strains, all belonging to a new yeast species, yet to be described, were also isolated.