Northern side of the Alps “INSIDE THE GLACIERS“ Project 2021

Alessio Romeo & Francesco Sauro

Revision A This document is neither contractual nor exhaustive, it may be modified and supplemented 30 April 2021 according to the evolution of the project and the integration of new participants.

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INSIDE THE GLACIERS

Northern side of the Alps

Switzerland () August-November 2021

CONTENTS :

1. INTRODUCTION : ...... 2 2. PROJECT HISTORY :...... 3 3. ORGANIZATION CHART : ...... 4 4. PLACES AND DATES : ...... 6 5. PROJECT OBJECTIVES : ...... 8 6. PROJECT PARTICIPANTS : ...... 9 7. OPERATIONS PROGRESS:...... 12 8. MICROBIAL ECOLOGY : ...... 13 9. NANOBIOCHEMISTRY : ...... 16 10. GLACIOLOGY : ...... 20 11. INTRA-GLACIAL EXPLORATION : ...... 21 12. DOCUMENTARY FILM : ...... 22 13. PARTNERS : ...... 23

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Northern side of the Alps

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1. INTRODUCTION :

The impact of climate change is particularly noticeable in the Alps due to its effects on glaciers and permafrost. The last few years have seen an increase in the number of landslides, debris flows and other collapses directly linked to the consequences of the melting of ice masses and normally frozen rocky areas. These geophysical phenomena can and will, at least in part, call into question certain human activities in the high alpine valleys as well as local urbanization. Beyond the ecological problems which are already dramatic in themselves, the consequences caused by these events are also economic and political. It is therefore vital to closely monitor the state of these glaciers and their evolution. Due to the large amount of glacial expanses present in its massifs, Valais, located in the heart of the Alps, is a territory particularly suitable for the study of alpine glaciers. It is for this reason that the organizers of the Inside The Glaciers project decided this year to carry out their investigations on various Valais glaciers. The study of ice caves in the temperate glaciers of the Planet began in the late ’80s with the union of mountaineering, caving and cave diving techniques. The exploration of this unknown world has opened new ways to scientific researches and it continues today including such aimed at biological and paleo-climatological studies. The cryo-karstic phenomenon (study of the evolution of glacier-caves) is directly connected with hydrogeology and physics of glaciers and this knowledge is still binding researchers in all glacial areas of Planet. Started in October 2014 with the first International Glacial Caving Camp on the Gorner Glacier (Zermatt – CH), the main objectives of the « INSIDE THE GLACIERS Project » Project are to collaborate with researchers and scientists to organize and manage exploration projects into the heart of glaciers, guaranteeing professional photographic and video documentation essential for subsequent media dissemination.

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2. PROJECT HISTORY :

The project INSIDE THE GLACIERS since its creation in 2014 is at the origin of many expeditions to different places on the Planet.

2014: • – Valais (Switzerland) • First International Glacier-Caving – Valais (Switzerland)

2015: • Abisso del Cenote – Dolomites (Italy) • 30th year of glacier-caving on Gorner Glacier – Valais (Switzerland)

2016: • Patagonia 2016 – Patagonia (Chile) • Grotta del Gelo al Mt. Etna - Sicily (Italy) • Cenote Abyss – Dolomites (Italy) • Gorner Glacier – Valais (Switzerland)

2017: • MaGPat – Patagonia (Argentina - Chile) • Gorner Glacier – Valais (Switzerland) • Greenland Ice cap – Kangerlussuaq (Groenland)

2018: • Inslandsis Groenlandais – Kangerlussuaq (Greenland) • Cenote Abyss – Dolomites (Italy)

2019: • Inylchek Glacier – Tian Shan (Kirghizstan)

2020: • Cenote Abyss – Dolomites (Italy) • Gorner Glacier – Valais (Switzerland) • Aletsch Glacier – Valais (Switzerland)

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3. ORGANIZATION CHART :

Organizers and communication managers :

Alessio Romeo Italian freelance photographer based in Tuscany. Explorer and caver from more than 30 years he is specialized in glacier-caves. Master degree in Geology with a thesis on the Gorner glacier (CH) with a study of morphological evolution of several glacier caves. Alessio Romeo has more than fifteen years of professional experience in adventure, cave and travel photography, an experience formed during more than 50 expeditions all over the World. Since 2013, he has been part of the team CAVES with different functions : Technical Assistant at the beginning, Cave-Photography teacher since 2016 and official photographer in 2019. In 2017 he start to collaborate as photographer also in the Planetary Geology training for astronauts PANGAEA (ESA). https://romeoalessiophotography.com

Francesco Sauro Italian explorer and speleologist, recognized by Time Magazine as one of the ten millennial that are having an impact in the future of science and planetary exploration. He is contracted Professor of Planetary Geology at the University of Bologna, with a peculiar interest in caves and subsurface exploration. With over twenty year of caving activity, Francesco Sauro has explored tens of kilometers of caves in any different lithology on Earth. In 2014 he won the Rolex Award for Enterprise with an exploration project in the Guyana highlands of South America. Since 2015 he has been selected as technical director of the course CAVES and PANGAEA organized by the European Space Agency for training astronauts (ESA, NASA, JAXA, RUSCOSMOS and CSA) for operational and for the study of human behavior purposes in harsh environments. http://labisso.blogspot.com

Logistics manager :

Didier Cassany Born and raised in the Alps (Haute-Savoie) near Valais, Didier Cassany works as mountain leader (UIMLA) and expedition leader (Alps, Himalaya). He is the author of several books for the prestigious Swiss Alpine Club and reports for French and Swiss mountain magazines. When he's not hiking, skiing or cycling in the Alps, he is in charge of the communication (social media strategy) for the British company Evergreencycling. https://www.evergreencycling.com

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French scientific research manager :

Roberta Brayner Chemical Engineer. Master of Science in satellite propulsion at the National Institute for Space Research (Brazil). Doctorate in Materials Chemistry at Paris Diderot University. Currently Associate Professor at the Interfaces, Processing, Organization and Systems Dynamics laboratory (ITODYS), University of Paris. Specialist in synthesis, characterization, application of nanoparticles, biosynthesis of nanoparticles using photosynthetic microorganisms, ecotoxicity of nanoparticles, detection by UV- Visible and Raman spectroscopy and electrochemistry of organic pollutants and heavy metals. Roberta Brayner participated in the Graal IV Expedition (2013) in Greenland for the identification of new species of microalgae and cyanobacteria resistant to cold and UV irradiation. Specialist in the characterization of cryoconite.

Italian scientific research manager :

Roberto Ambrosini Associate professor of ecology at the department of Environmental Science and Policy of the University of Milano. Roberto Ambrosini received the Msc in Natural Science and the PhD in Natural and Environmental Sciences from the University of Milano, Italy. He also received the BSc in statistics from the University of Milano Bicocca in 2007. His main scientific interests are the ecology of bird migration, the study of bird microbiomes, the ecology of glacier environments, and their contamination, particularly by microplastics. He has published more than 120 scientific papers on these topics in peer- reviewed journals.

Members of the French scientific team :

Nguyet Thanh Ha Duong Associate Professor at the laboratory ITODYS, University of Paris. Benoît Piro Professor at the laboratory ITODYS, University of Paris. Yann Sivry Associate Professor at the Institut de Physique du Globe de Paris (IPGP), Université de Paris. Nicolas Menguy Professor at the Institut de Minéralogie, de Physique des Matériaux et de Cosmochimie, Sorbonne University.

Members of the Italian scientific team :

Andrea Franzetti Associate Professor at the Department of Environmental and Earth Sciences of the University of Milan - Bicocca. Davide Fugazza Associate Professor at the Department of Environmental Science and Policy of the University of Milan. INSIDE THE GLACIERS insidetheglaciers.wordpress.com Alessio.Romeo: [email protected] Francesco Sauro: [email protected] 5 INSIDE THE GLACIERS

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4. PLACES AND DATES :

This project will take place over a period from August to November 2021 in the , more precisely in the canton of Valais, according to a traveling route that will allow you to explore and study from inside and outside the following glaciers: Allalin Glacier Corbassière Glacier

(Saastal, Saas Almagell) (Val de Bagnes, Bagnes) Ferpècle Glacier Lang Glacier

(Val d’Hérens, Evolène) (Lötschental, ) Moiry Glacier Plaine Morte Glacier

(Val de Moiry, Grimentz) (, Crans-Montana)

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Zinal Glacier Aletsch Glacier

(Val d’Anniviers, ) (Fieschertal, Brig)

Geographical distribution of the different objective glaciers of this project :

Répartition des glaciers retenus (tous situés dans le Valais) :

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5. PROJECT OBJECTIVES :

The objectives of the Northern side of the Alps project are as follows :

• Carry out glaciological surveys on glaciers and in contact caves where the simultaneous presence of water and air has a significant impact on the process of ice melting and collapse. This work will be carried out by Doctor Davide Fugazza of the University of Milan in collaboration with the organizers (Alessio Romeo and Francesco Sauro) and the caving technicians, through field investigations, surveys with high resolution drones and satellite images. • Perform samples of bacteria present on glaciers as well as in cavities in order to establish connections between these different groups. This study will be carried out by Professors Andrea Franzetti of the University of Milan-Bicocca and Roberto Ambrosini of the University of Milan. • Take samples in order to study mineral interactions / extremophilic microorganisms, verify the presence of pollutants (organic elements, heavy metals, nanoparticles and microplastics) and study the resistance of these extremophilic microorganisms in the presence of these pollutants. These investigations will be led by Doctor Roberta Brayner and Professors Nguyêt- Than Ha Duong, Benoît Piro and Yann Sivry from the University of Paris as well as by Professor Nicolas Menguy from Sorbonne University. • Explore endoglacial cavities in order to better understand endoglacial hydrology. • Shoot the sequences of a film recounting the work and the explorations carried out during the project.

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6. PROJECT PARTICIPANTS :

The Northern side of the Alps project led by explorer and photographer Alessio Romeo and Professor Francesco Sauro will rely on the active participation of universities, research centers, companies (for 3d digitization) and the following associations :

Universities :

Università degli studi di Milano (Italy) www.unimi.it

Università degli studi di Milano Bicocca (Italy) www.unimib.it

Université de Paris (France) u-paris.fr

Sorbonne Université (France) www.sorbonne-universite.fr

Research centers :

Centre National de la Recherche Scientifique (France) www.cnrs.fr

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DISAT (Italy) www.disatmicrobiology.it

Institut de Minéralogie, de Physique des Matériaux et de Cosmochimie (France) www.umpmc.upmc.fr

Institut de Physique du Globe de Paris (France) www.ipgp.fr

Interface Traitements Organisation et Dynamique des Systemes (France) www.itodys.univ-paris-diderot.fr

Monitoring and Management of Microbial Resources (Italie) www.m3r.it

Companies:

Leica Geosystems (France) https://leica-geosystems.com

Virtual Geographic Agency (Italy) www.vigea.it

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Associations :

Associazione Speleologia Progetto Supramonte (Italy) www.universirius.com/a-s-pro-s

LA VENTA Esplorazioni geografiche (Italy) www.laventa.it

Mille et un Pas sous la Glace (France)

Progetto Speleologia Glaciale (Italy) progettospleologiaglaciale.org

Regards sur l’Aventure (France) www.rsla.fr

SPELE'ICE Explorations (France) www.speleice.fr

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7. OPERATIONS PROGRESS:

The schedule for the reconnaissance operation (August) and the effective expeditions (between September and November 2021) has not yet been defined since it will be the weather forecast and the availability of the various groups and associations that will determine it.

Pre-expedition (August 2021)

In August, the listed glaciers will be visited by several teams of speleologists and researchers in order to determine the logistics, their morphological configuration and the presence of points of interest for the purposes of the project. The summer period is probably the most suitable, in particular for biological and microbiological samples which involve the presence of liquid water both on the surface and inside the glaciers. In agreement with researchers from the University of Bicocca we will take advantage of the hottest period both to start sampling and to train technicians / speleologists on sampling methods, which especially inside the glacier, must be carried out by experienced and competent persons in underground environment. It is expected that, for each glacier, the work will not take more than 3 / 4 days and that each group (made up of both caving technicians and scientists) tries to move as lightly as possible. If necessary, the use of helicopters is envisaged, in particular on the most difficult to access glaciers and where the work requires heavier logistics or cumbersome technical / scientific equipment.

Expedition (September-November 2021)

In the period between September and October, we find the ideal conditions for the exploration of glacial cavities and contact caves, where speleologists can carry out topographies of internal spaces, photographic documentation and video and, of course, on the instruction of scientists, also sampling of various types.As already specified, one of the objectives of the project is the mapping and documentation of cavities in contact between ice and rock. Laser scans for 3D reconstruction of the environments could provide interesting seasonal and annual information on the variation of these environments affected by the melting of the ice due to both the passage of water and the passage of air currents caused by the presence of multiple entrances. This phenomenon is, at this historical moment, one of the processes which accelerates the most the disappearance of the masses of ice and creates situations of instability of them which becomes a potential danger for the downstream valleys.

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8. MICROBIAL ECOLOGY :

Glaciers are not lifeless environments but harbor complex communities of microorganisms composed mostly of bacteria. Their growth and metabolism can have a major impact on melting ice as well as on maintaining essential ecological functions for downstream ecosystems.

Cold and isolated environments present similar environmental conditions and are linked by long-distance transport mechanisms. These characteristics make them ideal for the study of microbial biogeographic processes and other basic ecological processes that determine the structure and functions of microbial communities.

Cryoconite holes are small ponds full of meltwater and with a sediment on the bottom present on the surface of most glaciers. Although they are characterized by extreme conditions such as low temperatures and high solar irradiance, they host bacterial communities with high taxonomic and functional biodiversity. It has been supposed that such high diversity could be due to the high versatility of some of the most abundant populations. In a previous investigation, we showed that these supraglacial communities exhibit high functional biodiversity since they exploit organic matter both as energy and carbon source, and use both oxygenic and anoxygenic photosynthesis with pure autotrophic and mixotrophic lifestyles. We also demonstrated the importance of light in driving carbon fluxed on the glacier surface.

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Figure 1 (a–f) Light micrographs of algae found at the glacier. (a–c) Filaments of Ancylonema nordenskioeldii. (d,e) unicellular Mesotaenium berggrenii. (f) red cyst of putative Sanguina nivaloides (g) Aerial view of the Morteratsch glacier tongue acquired in September 2016 from an Unmanned Aerial Vehicle (UAV.

However, due to the lack of expression studies on these environments, it is currently unknown whether these metabolic functions are all concomitantly active, or there is a temporal pattern of expression, which, in turn, may contribute to the whole functional biodiversity. To investigate this hypothesis, we propose to regularly collect samples from i) cryoconite holes and from ii) endoglacial sediment along typical summer consecutive days on the Greenland glacier and to use shotgun metatransctriptomics sequencing to investigate the expression of the main metabolic functions. Preliminary data actually suggested that, on the surface of the glacier (cryoconite holes), carbon and energy metabolisms had different patterns of expression along the day. Particularly, the taxa expressing CO2 fixation are extremely variable along the day and span for phototrophic to chemosynthetic organisms.

Figure 2 Bacteria colonies from 100 µL of snow grown at 4°C in light conditions. Snow was collected on the Iver glacier (Chile, ~ 4200 m a.s.l.). INSIDE THE GLACIERS insidetheglaciers.wordpress.com Alessio.Romeo: [email protected] Francesco Sauro: [email protected] 14 INSIDE THE GLACIERS

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Figure 3 Oxygenated and anoxygenic photosynthesis process.

If confirmed in the proposed project, these results indicates functional redundancy likely enhances that microbial ecosystems the ecosystem stability in such extreme environments. Furthermore, despite the fact that supraglacial environments are considered as a whole as oxic, there is recent evidence that, at the microscopic scale, anoxic conditions are established in supraglacial sediments which allow the growth of anaerobic microorganisms, thus increasing the taxonomic and functional biodiversity of these environments. For this reason we propose to study the vertical profiles of oxygen and hydrogen at the micrometric scale in cryoconite using microsensors.

This project would also aim to expand knowledge of the endoglacial environment in order to determine whether time-dependent expression profiles are also present due to microbial circadian rhythms.

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9. NANOBIOCHEMISTRY :

This is a scientific study at the interface between chemistry, physics and microbiology with multiple objectives: (i) it is primarily to study mineral interactions / extremophilic microorganisms (cryoconite), (ii) verify the presence of organic pollutants and heavy metals, (iii) verify the presence of nanoparticles and microplastics, and finally (iv) study the resistance of these extremophilic microorganisms in the presence of these pollutants.

Anthropogenic vs geogenic nanoparticles :

While climatic and geological phenomena were for a long time the main drivers of the transformation of land surfaces, we are studying how Man, at the time of the Anthropocene, is at the origin of the majority of the transformations that affect the area. critical. In this context, the study of the biogeochemical cycles of metallic elements and more specifically of their evolution under the action of anthropogenic forcings is focusing the growing attention of scientists.

Among anthropogenic NPs, we can distinguish those manufactured from those released unintentionally into the environment: from vehicle exhaust fumes, industrial activities (steelworks and foundries, construction) or domestic activities. Emissions from traffic and the combustion of fossil fuels generally produce carbon-based NPs (soot or carbon black, but also carbon nanotubes, fullerenes and carbonaceous iron oxides). Metal mining and refineries produce NPs of metals and metal oxides such as NPs of Ag, Pb, Se, Sb and Zn. Large-scale demolitions of old buildings (often constructed with toxic materials) have been proven to produce NPs, such as breathable asbestos fibers or lead molecules, which can also be transported great distances.

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Figure 4 (a) Eruption of Eyjafjallajökull volcano, Iceland, April 2010 (image courtesy of Carolyn Kaster); (b) SEM image showing aggregates of NPs from volcanic ash from Eyjafjallajökull volcano (image courtesy from Gislason et al. 2011); (c) and (d) manufactured gold nanoparticles; (e) TEM images of Aquaspirillum magnetotacticum bacterium showing magnetosomes (iron oxide granules); (f) SEM images of SEM silica frustules (cell walls with silicate NPs). Images courtesy of Dr. Dennis Kunkel (www.visualsunlimited.com).

At the same time, the problems concerning the preservation of drinking water reserves, natural environments and biodiversity have led governments to adopt drastic measures in terms of environmental policy.

Analysis by absorption and emission spectroscopy of the metabolism of microorganisms present in the ice cap and search for new fluorescent proteins for biotechnology :

UV and visible absorption and emission spectroscopies are powerful analytical techniques. These methods are fast, sensitive and easy to implement. In addition, technical innovations provide portable devices necessary for scientific expeditions. For example, UV-visible absorption spectroscopy has made it possible to study the complexation of heavy metals such as cadmium or lead by glutathione or phytochelatins, natural molecules present in plants. The detection of other metals such as cesium or lead can be easily achieved by fluorescence in the presence of synthetic (calixarene) or natural (norbadione) molecules. The metabolism of extremophilic microorganisms can thus be easily studied and analyzed to understand the adaptation mechanisms of these organisms under extreme conditions, or in the presence of organic pollutants or heavy metals. The spectral footprints of these organisms will be different depending on their living environment. The absorption and emission spectroscopies will be complementary to the EGOFET technique, developed by the BiOSS team in the laboratory.

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Finally, extremophilic microorganisms have proteins which are particularly appreciated for biotechnology. These extremozymes are stable under acidic or basic pH conditions, high pressure and / or high ionic strength, which allows the survival of these organisms. Cold-adapted proteins find use in detergent development or food processing, and could be exploited in industry as anti-freeze products. Certain proteins are fluorescent with important luminescence properties (stability, quantum yield, wide range of emission in the visible), they can then be used in imaging or as sensors.

Magnetotactic bacteria are a morphologically, phylogenetically and physiologically diverse group of aquatic prokaryotes that are found in many parts of the world. They are found in chemically stratified sediments or water columns. These bacteria live in extremely varied environments in terms of salinity (saline, brackish or freshwater), temperature (low to high and in extreme conditions, great depth or even high pH). These microorganisms have the particularity of having one or more intracellular chains of magnetic crystals made up of magnetite (Fe3O4) or greigite (Fe3S4). These chains of nanoparticles give these bacteria the ability to move along the field lines, thus facilitating their migration to areas of low oxygen concentration (microaerobic) where their growth is optimal. This passive alignment of bacteria associated with chemotaxis is called aero-magnetotaxis. Two publications mention the existence of psychrophilic magnetotactic bacteria, ie making their entire life cycle in cold biotopes. A Brazilian team has reported bacteria belonging to the class of alpha-proteobacteria living in waters with a temperature below 1 ° C (King Georges Island on the Antarctic continent 62.05 S, 58.30 W). Two researchers from BIAM / CEA-CNRS-AMU, associated with speleologists from Clermont-Ferrand have identified magnetotactic bacteria living in fresh water at 4 ° C in Creux de Soucy. This work indicates that the biomineralization of bacterial magnetic nanoparticles can take place in cold ecosystems. However, the processes of their formation in such environments are unknown. To date, there is no research regarding the presence of such bacteria in the northern regions. The observation of such microorganisms in polar environments with a magnetic field whose inclination and intensity are very particular given the proximity of the Earth's magnetic South Pole could contribute to a better knowledge of their biodiversity and a better understanding of the aero-magnetotaxis.

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Recent optimizations of techniques for collecting, concentrating and observing magnetotactic microorganisms applied to aquatic environments with unique physicochemical properties have made it possible to describe new taxa, nanoparticles, physiologies and biological processes. The development of these technological innovations, easily deployable in the field, in the aquatic environments of Greenland will certainly lead to exciting discoveries.

Figure 5 Electron microscopy images showing two magnetotactic bacteria with the ability to biomineralize magnetic iron nanoparticles and phosphorus-rich granules.

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10. GLACIOLOGY :

Glaciers are sensitive barometers of climate change, constantly and quickly expanding and contracting in response to changes in air temperature and snowfall. The ongoing climate warming dramatically accelerated at the end of the 20th Century. Indeed, many mountain ranges of the Planet have lost a significant portion of their glacierized areas over the past 150 years and a rapid acceleration of glacier retreat has been recorded over the past two decades. Besides the large areal variation, the surface of glaciers across the planet has undergone important physical changes: the areal expansion of supraglacial debris cover, deposition of mineral dust and “black carbon” and the growth of glacier algae have been reported in major mountain ranges. The combination of these phenomena, defined as “glacier darkening”, has a fundamental impact on the energy balance of glaciers themselves, increasing the absorption of solar radiation and enhancing glacier melting o the ice. The Greenland ice sheet is one of the areas of the Earth most affected by this darkening process and although its origin, effects and characteristics have recently been investigated, many questions remain open. But this process also takes place on our doorstep in the Alps .

This project is aimed at using high-resolution UAV surveys with multispectral sensors to disentangle the different contributions of biotic and abiotic components to the darkening of the Greenland Ice Sheet. UAV data will also be coupled with measurements from the Sentinel-2 satellite to understand mechanisms of dust deposition and the role of algal blooms in enhancing surface melting.

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11. INTRA-GLACIAL EXPLORATION :

The complexity of glacial and subglacial explorations requires techniques combining caving and mountaineering. Knowledge of these very hostile environments is essential to succeed in such missions.The major objective of subglacial exploration is to try to elucidate the mode of circulation of meltwater at great depth. How far do these, grouped together in "bédières" (surface rivers) and captured by "moulins" (glacial chasms), sink into the glacier ? Does the water go to the bottom of the crevasse or is there a depth threshold ? Are there any galleries in the shape of a collector ? How are the caves evolving ? Does the type of ice, bullous or not, intervene in the morphology of the cryo-karstic cavity ? There are many more questions for ice scientist-explorers. The interest of this research lies in the applications which, for some, have already been undertaken (catchments, extraction of fresh water in glacial and oceanic environments). If the Greenlandic ice sheet, by its strong potential, constitutes the zone of predilection for the exploration of the mills, it remains particularly interesting to explore other glacial zones in order to be able to establish comparisons on the development of these cryo- karst formations. Beyond the exploration itself and the topography of the glacial chasms, an essential task of the explorers will be to put their mastery of speleological and alpine techniques at the service of the scientists in order to accompany and supervise them inside the cavities or, where appropriate, take samples in their place.

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12. DOCUMENTARY FILM :

Associating expeditions with quality video and photographic documentation is a constant requirement. The precision and the high level of quality of the documentation produced will guarantee a strong media potential which can be exploited through scientific publications and popularization. Photographic and video coverage will be provided by several professionals.

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13. PARTNERS :

[Partner logos]

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