Nicolas Walter

AstRoMap European Astrobiology Roadmap

Abstract The European AstRoMap project (supported by the European Commission Seventh Framework Programme) surveyed the state of the art of astrobiology in Europe and beyond and produced the first European roadmap for astrobiology research. In the context of this roadmap, astrobiology is understood as the study of the origin, evolution, and distribution of life in the context of cosmic evolution; this includes habitability in the Solar System and beyond. The AstRoMap Roadmap identifies five research topics, specifies several key scientific objectives for each topic, and suggests ways to achieve all the objectives. The five AstRoMap Research Topics are • Research Topic 1: Origin and Evolution of Planetary Systems • Research Topic 2: Origins of Organic Compounds in Space • Research Topic 3: Rock-Water-Carbon Interactions, Organic Synthesis on Earth, and Steps to Life • Research Topic 4: Life and Habitability • Research Topic 5: Biosignatures as Facilitating Life Detection It is strongly recommended that steps be taken towards the definition and implementation of a European Astrobiology Platform (or Institute) to streamline and optimize the scientific return by using a coordinated infrastructure and funding system. Key Words: Astrobiology roadmap—Europe—Origin and evolution of life—Habitability—Life detection—Life in extreme environments. Astrobiology 16, 201–243.

The responses of an anaerobic microorganism, Yersinia intermedia MASE-LG-1 to individual and combined simulated Martian stresses

The limits of life of aerobic microorganisms are well understood, but the responses of anaerobic microorganisms to individual and combined extreme stressors are less well known. Motivated by an interest in understanding the survivability of anaerobic microorganisms under Martian conditions, we investigated the responses of a new isolate, Yersinia intermedia MASE-LG-1 to individual and combined stresses associated with the Martian surface. This organism belongs to an adaptable and persistent genus of anaerobic microorganisms found in many environments worldwide. The effects of desiccation, low pressure, ionizing radiation, varying temperature, osmotic pressure, and oxidizing chemical compounds were investigated. The strain showed a high tolerance to desiccation, with a decline of survivability by four orders of magnitude during a storage time of 85 days. Exposure to X-rays resulted in dose-dependent inactivation for exposure up to 600 Gy while applied doses above 750 Gy led to complete inactivation. The effects of the combination of desiccation and irradiation were additive and the survivability was influenced by the order in which they were imposed. Ionizing irradiation and subsequent desiccation was more deleterious than vice versa. By contrast, the presence of perchlorates was not found to significantly affect the survival of the Yersinia strain after ionizing radiation. These data show that the organism has the capacity to survive and grow in physical and chemical stresses, imposed individually or in combination that are associated with Martian environment. Eventually it lost its viability showing that many of the most adaptable anaerobic organisms on Earth would be killed on Mars today.

Anaerobic microorganisms in astrobiological analogue environments: from field site to culture collection

Astrobiology seeks to understand the limits of life and to determine the physiology of organisms in order to better assess the habitability of other worlds. To successfully achieve these goals we require microorganisms from environments on Earth that approximate to extraterrestrial environments in terms of physical and/or chemical conditions. The most challenging of these environments with respect to sample collection, isolation and cultivation of microorganisms are anoxic environments. In this paper, an approach to this challenge was implemented within the European Unions MASE (Mars Analogues for Space Exploration) project. In this review paper, we aim to provide a set of methods for future field work and sampling campaigns. A number of anoxic environment based on characteristics that make them analogous to past and present locations on Mars were selected. They included anoxic sulphur-rich springs (Germany), the salt-rich Boulby Mine (UK), a lake in a basaltic context (Iceland), acidic sediments in the Rio Tinto (Spain), glacier samples (Austria) and permafrost samples (Russia and Canada). Samples were collected under strict anoxic conditions to be used for cultivation and genomic community analysis. Using the samples, a culturing approach was implemented to enrich anaerobic organisms using a defined medium that would allow for organisms to be grown under identical conditions in future physiological comparisons. Anaerobic microorganisms were isolated and deposited with the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) culture collection to make them available to other scientists. In MASE, the selected organisms are studied with respect to survival and growth under Mars relevant stresses. They are artificially fossilized and the resulting biosignatures studied and used to investigate the efficacy of life detection instrumentation for planetary missions. Some of the organisms belong to genera with medical and environmental importance such as Yersinia spp., illustrating how astrobiology field research can be used to increase the availability of microbial isolates for applied terrestrial purposes.

THESEUS: The European research priorities for human exploration of space

Today, the exploration of space remains one of the most stimulating and exciting areas of scientific research and technological development. One of the objectives for the next 10 years is to create, and then implement, a long-term plan for the robotic and human exploration of the solar system, with Mars and the Moon as first targets. To undertake such a future mission requires major efforts of global and interdisciplinary cooperation between scientific, industrial, and legislative parties. This was recently highlighted in the report by the Committee on Human Spaceflight of the National Academy of Sciences of the United States of America. Past space missions in Earth orbit have demonstrated that humans can survive and work in space for durations of up to several months and return to Earth with relatively limited health consequences. However, there are pending technological, medical, and psychological issues to be solved before adventuring in longer and more distant space missions, such as those envisioned in a space exploration program. For example, protection against ionizing radiation, problems with lunar and Martian dust, a reliable closed life-support system in transit and on the surface, psychological issues such as those affecting cognition, behavior, and performance during and after long-duration space travel, general metabolic disturbances such as prevention of bone loss and muscle atrophy, and the potential irreversibility of these changes, as well as balance and coordination as the main limiting factors for a manned mission to Mars. Similarly, the known enhanced infectious disease risks as humans travel and live within these stressful, confined environments, require special attention. Furthermore, technological breakthroughs, primarily in lifesupport systems and recycling technologies are required to reduce the costs of these expeditions to more acceptable levels. Solving such issues will need creative scientific and technological approaches relevant to clinical and industrial applications here on Earth. In the United States, NASA Human Research Program implements a constantly evolving Human Research Roadmap that represents a “risk reduction strategy for human space exploration.” In Europe, European Space Agency (ESA) supported several preliminary studies enabling the definition of priorities to prepare for interplanetarymanned exploration missions, such as the HUMEX (study on the Survivability and Adaptation of Humans to long-duration Exploratory Missions) or FIPES (Facility for Integrated Planetary Exploration Simulation). ESA has also developed a comprehensive and targeted ground program to validate effective countermeasures through the bed rest studies, including the testing of the use of artificial gravity applied through a short-arm centrifuge. However, Europe had not developed its own human exploration roadmap. On the basis of the current knowledge, an action plan approved by the European scientific and industrial communities, and relying on ESA programs, integrating the expertise of non-ESA member states as well as of the European Eastern countries is needed. In 2012, the European Union funded the project THESEUS (Towards Human Exploration of Space: A European Perspective, supported by the EU Seventh Framework Program for Research and Technology Development). The goals of THESEUS were to develop an integrated life sciences research roadmap enabling European human space exploration, in synergy with the ESA strategy, taking advantage of the expertise available in Europe, and identifying the potential of non-space applications and development. As any human exploration initiative can only be conceived of as a well-coordinated series of programs at the international level, the issue of international cooperation and coordination with other nations, agencies, and programs has been central to the establishment of such a roadmap. Space agencies are currently jointly discussing their plans for exploring the Moon and Mars. Any research roadmap enabling human exploration of these targets should, both, incorporate these plans in its making, and influence such plans to ensure that basic and applied research priorities as defined and expressed by the relevant communities are addressed in a consistent manner by these agencies. To enable this, the concerned space agencies were represented as observers in THESEUS and international experts were invited to participate in the expert groups. THESEUS had three objectives: (i) identify disciplinary research priorities, (ii) focus on fields with high-terrestrial application potential, and (iii) build a European network as the core of this strategy. These three objectives were tackled through five tightly interrelated clusters involving 137 international scientists:

Beyond Chloride Brines: Variable Metabolomic Responses in the Anaerobic Organism Yersinia intermedia MASE-LG-1 to NaCl and MgSO4 at Identical Water Activity

Growth in sodium chloride (NaCl) is known to induce stress in non-halophilic microorganisms leading to effects on the microbial metabolism and cell structure. Microorganisms have evolved a number of adaptations, both structural and metabolic, to counteract osmotic stress. These strategies are well-understood for organisms in NaCl-rich brines such as the accumulation of certain organic solutes (known as either compatible solutes or osmolytes). Less well studied are responses to ionic environments such as sulfate-rich brines which are prevalent on Earth but can also be found on Mars. In this paper, we investigated the global metabolic response of the anaerobic bacterium Yersinia intermedia MASE-LG-1 to osmotic salt stress induced by either magnesium sulfate (MgSO4) or NaCl at the same water activity (0.975). Using a non-targeted mass spectrometry approach, the intensity of hundreds of metabolites was measured. The compatible solutes L-asparagine and sucrose were found to be increased in both MgSO4 and NaCl compared to the control sample, suggesting a similar osmotic response to different ionic environments. We were able to demonstrate that Yersinia intermedia MASE-LG-1 accumulated a range of other compatible solutes. However, we also found the global metabolic responses, especially with regard to amino acid metabolism and carbohydrate metabolism, to be salt-specific, thus, suggesting ion-specific regulation of specific metabolic pathways.

Mineralization and Preservation of an extremotolerant Bacterium Isolated from an Early Mars Analog Environment

The artificial mineralization of a polyresistant bacterial strain isolated from an acidic, oligotrophic lake was carried out to better understand microbial (i) early mineralization and (ii) potential for further fossilisation. Mineralization was conducted in mineral matrixes commonly found on Mars and Early-Earth, silica and gypsum, for 6 months. Samples were analyzed using microbiological (survival rates), morphological (electron microscopy), biochemical (GC-MS, Microarray immunoassay, Rock-Eval) and spectroscopic (EDX, FTIR, RAMAN spectroscopy) methods. We also investigated the impact of physiological status on mineralization and long-term fossilisation by exposing cells or not to Mars-related stresses (desiccation and radiation). Bacterial populations remained viable after 6 months although the kinetics of mineralization and cell-mineral interactions depended on the nature of minerals. Detection of biosignatures strongly depended on analytical methods, successful with FTIR and EDX but not with RAMAN and immunoassays. Neither influence of stress exposure, nor qualitative and quantitative changes of detected molecules were observed as a function of mineralization time and matrix. Rock-Eval analysis suggests that potential for preservation on geological times may be possible only with moderate diagenetic and metamorphic conditions. The implications of our results for microfossil preservation in the geological record of Earth as well as on Mars are discussed.

Lack of correlation of desiccation and radiation tolerance in microorganisms from diverse extreme environments tested under anoxic conditions

Abstract Four facultative anaerobic and two obligate anaerobic bacteria were isolated from extreme environments (deep subsurface halite mine, sulfidic anoxic spring, mineral-rich river) in the frame MASE (Mars Analogues for Space Exploration) project. The isolates were investigated under anoxic conditions for their survivability after desiccation up to 6 months and their tolerance to ionizing radiation up to 3000 Gy. The results indicated that tolerances to both stresses are strain-specific features. Yersinia intermedia MASE-LG-1 showed a high desiccation tolerance but its radiation tolerance was very low. The most radiation-tolerant strains were Buttiauxella sp. MASE-IM-9 and Halanaerobium sp. MASE-BB-1. In both cases, cultivable cells were detectable after an exposure to 3 kGy of ionizing radiation, but cells only survived desiccation for 90 and 30 days, respectively. Although a correlation between desiccation and ionizing radiation resistance has been hypothesized for some aerobic microorganisms, our data showed that there was no correlation between tolerance to desiccation and ionizing radiation, suggesting that the physiological basis of both forms of tolerances is not necessarily linked. In addition, these results indicated that facultative and obligate anaerobic organisms living in extreme environments possess varied species-specific tolerances to extremes.