SPECTROModule: A modular in-situ spectroscopy platform for exobiology and space sciences

The evolution of the solar system and the origin of life remain some of the most intriguing questions for humankind. Addressing these questions experimentally is challenging due to the difficulty of mimicking environmental conditions representative for Early Earth and/or space conditions in general in ground-based laboratories. Performing experiments directly in space offers the great chance to overcome some of these obstacles and to possibly find answers to these questions. Exposure platforms in Low Earth Orbit (LEO) with the possibility for long-duration solar exposure are ideal for investigating the effects of solar and cosmic radiation on various biological and non-biological samples. Up to now, the Exobiology and space science research community has successfully made use of the International Space Station (ISS) via the EXPOSE facility to expose samples to the space environment with subsequent analyses after return to Earth. The emerging small and nanosatellite market represents another opportunity for astrobiology research as proven by the robotic O/OREOS mission, where samples were monitored in-situ, i.e. in Earth orbit. In this framework, the European Space Agency is developing a novel Exobiology facility outside the ISS. The new platform, which can host up to four different experiments, will combine the advantages of the ISS (long-term exposure, sample return capability) with near-real-time in-situ monitoring of the chemical/biological evolution in space. In particular, ultraviolet–visible (UV–Vis) and infrared (IR) spectroscopy were considered as key non-invasive methods to analyse the samples in situ. Changes in the absorption spectra of the samples developing over time will reveal the chemical consequences of exposure to solar radiation. Simultaneously, spectroscopy provides information on the growth rate or metabolic activities of biological cultures. The first quartet of experiments to be performed on-board consists of IceCold, OREOcube and Exocube (dual payload consisting of ExocubeChem and ExocubeBio). To prepare for the development of the Exobiology facility, ground units of the UV–Vis and IR spectrometers were studied, manufactured and tested as precursors of the flight units. The activity led to a modular in-situ spectroscopy platform able to perform different measurements (e.g. absorbance, optical density, fluorescence measurements) at the same time on different samples. We describe here the main features of the ground model platform, the verification steps, results and approach followed in the customization of commercial–off-the-shelf (COTS) modules to make them suitable for the space environment. The environmental tests included random and shock vibration, thermal vacuum cycles in the range −20 °C to +40 °C and irradiation of the components with a total dose of 1800 rad (18 Gy). The results of the test campaign consolidated the selection of the optical devices for the Exobiology Facility. The spectroscopic performance of the optical layout was tested and benchmarked in comparison with state-of-the-art laboratory equipment and calibration standards showing good correlation. This includes spectra of samples sets relevant for the flight experiments and a performance comparison between the SPECTROModule ground model and state-of-the-art laboratory spectrometers. Considering the large number of samples and different types of optical measurements planned on-board the ISS, the main outcome was the implementation of an LED-photodiode layout for the optical density and fluorescence measurements of IceCold (42 samples) and ExocubeBio (111 samples); while the UV–Vis spectrometer will be mainly focused on the change of the absorption spectra of the 48 samples of OREOcube.The ExocubeChem samples (in total 48) will be analysed by infrared spectroscopy. The ground platform supports the establishment of analogue research capabilities able to address the long-term objectives beyond the current application

Metallosphaera sedula on a Mission – mimicking Mars in frames of the Tanpopo 4 mission

With future long-term space exploration and life detection missions on Mars, understanding the microbial survival beyond Earth as well as the identification of past life traces on other planetary bodies becomes increasingly important. The series of the Tanpopo space mission experiments target a long-term exposure (one to three years) of microorganisms on the KIBO Module of the International Space Station (ISS) in the low Earth orbit (LEO) (Kawaguchi et al., 2020; Ott et al., 2020). In the search for possible past and/or present microbial life on Mars, metallophilic archaeal species are of a special interest due to their inherent extraordinary characteristics. Chemolithotrophic archaea (e.g., from the order Sulfolobales) employ a number of ancient metabolic pathways to extract energy from diverse inorganic electron donors and acceptors. Metallosphaera sedula, an iron- and sulfur-oxidizing chemolithotrophic archaeon, which flourishes under hot and acidic conditions (optimal growth at 74°C and pH 2.0), was cultivated on genuine extraterrestrial minerals (Milojevic et al., 2019; Milojevic et al., 2021) as well as synthetic Martian materials (Kölbl et al., 2017). In all cases, M. sedula cells were able to utilize given mineral materials as the sole energy source for cellular growth and proliferation. During the growth of M. sedula cells on these materials, a natural mineral impregnation and encrustation of microbial cells was achieved, followed by their preservation under the conditions of desiccation (Kölbl et al. 2020). Our studies indicate that this archaeon, when impregnated and encrusted with minerals, withstand long-term desiccation and can be even recovered from the dried samples to the liquid cultures (Kölbl et al., 2020). The achieved preservation of desiccated M. sedula cells facilitated our further survivability studies with this desiccated microorganism under simulated Mars-like environmental conditions and during the Tanpopo-4 space exposure experiment. [...

Molecular response of Deinococcus radiodurans exposed to vacuum conditions of Low Earth Orbit

The polyextremophile, gram-positive bacterium Deinococcus radiodurans is able to withstand harsh conditions of real and simulated outer space environment, e.g., extreme temperature fluctuations, desiccation, UV radiation, and ionizing radiation. A long-term space exposure of D. radiodurans has been performed in exposure experiments at low Earth orbit in frames of the Tanpopo orbital mission aiming to investigate the possibility of interplanetary transfer of life. Although it is important to analyse the impact of space environmental factors simultaneously, it is also crucial to investigate these factors separately under controlled conditions in order to decipher fundamental response mechanisms involved

Proteomic and Metabolomic Profiling of Deinococcus radiodurans Recovering After Exposure to Simulated Low Earth Orbit Vacuum Conditions

The polyextremophile, gram-positive bacterium Deinococcus radiodurans can withstand harsh conditions of real and simulated outer space environment, e.g., UV and ionizing radiation. A long-term space exposure of D. radiodurans has been performed in Low Earth Orbit (LEO) in frames of the Tanpopo orbital mission aiming to investigate the possibility of interplanetary life transfer. Space vacuum (10-4–10-7 Pa) is a harmful factor, which induces dehydration and affects microbial integrity, severely damaging cellular components: lipids, carbohydrates, proteins, and nucleic acids. However, the molecular strategies by which microorganisms protect their integrity on molecular and cellular levels against vacuum damage are not yet understood. In a simulation experiment, we exposed dried D. radiodurans cells to vacuum (10-4–10-7 Pa), which resembles vacuum pressure present outside the International Space Station in LEO. After 90 days of high vacuum exposure, survival of D. radiodurans cells was 2.5-fold lower compared to control cells. To trigger molecular repair mechanisms, vacuum exposed cells of D. radiodurans were recovered in complex medium for 3 and 6 h. The combined approach of analyzing primary metabolites and proteins revealed important molecular activities during early recovery after vacuum exposure. In total, 1939 proteins covering 63% of D. radiodurans annotated protein sequences were detected. Proteases, tRNA ligases, reactive oxygen species (ROS) scavenging proteins, nucleic acid repair proteins, TCA cycle proteins, and S-layer proteins are highly abundant after vacuum exposure. The overall abundance of amino acids and TCA cycle intermediates is reduced during the recovery phase of D. radiodurans as they are needed as carbon source. Furthermore, vacuum exposure induces an upregulation of Type III histidine kinases, which trigger the expression of S-layer related proteins. Along with the highly abundant transcriptional regulator of FNR/CRP family, specific histidine kinases might be involved in the regulation of vacuum stress response. After repair processes are finished, D. radiodurans switches off the connected repair machinery and focuses on proliferation. Combined comparative analysis of alterations in the proteome and metabolome helps to identify molecular key players in the stress response of D. radiodurans, thus elucidating the mechanisms behind its extraordinary regenerative abilities and enabling this microorganism to withstand vacuum stress

К определению поверхностного натяжения, объема и площади криволинейной поверхности по форме сидячих пузырьков или висячих капель

The multi-user facility EXPOSE-E was designed by the European Space Agency to enable astrobiology research in space (low-Earth orbit). On 7 February 2008, EXPOSE-E was carried to the International Space Station (ISS) on the European Technology Exposure Facility (EuTEF) platform in the cargo bay of Space Shuttle STS-122 Atlantis. The facility was installed at the starboard cone of the Columbus module by extravehicular activity, where it remained in space for 1.5 years. EXPOSE-E was returned to Earth with STS-128 Discovery on 12 September 2009 for subsequent sample analysis. EXPOSE-E provided accommodation in three exposure trays for a variety of astrobiological test samples that were exposed to selected space conditions: either to space vacuum, solar electromagnetic radiation at > 110nm and cosmic radiation (trays 1 and 3) or to simulated martian surface conditions (tray 2). Data on UV radiation, cosmic radiation, and temperature were measured every 10 s and downlinked by telemetry. A parallel mission ground reference (MGR) experiment was performed on ground with a parallel set of hardware and samples under simulated space conditions. EXPOSE-E performed a successful 1.5-year mission in space

Metabolomic Profile of the Fungus Cryomyces antarctiucus Under Simulated Martian and Space Conditions as Support for Life-Detecion Missions on Mars

The identification of traces of life beyond Earth (e.g., Mars, icy moons) is a challenging task because terrestrial chemical-based molecules may be destroyed by the harsh conditions experienced on extraterrestrial planetary surfaces. For this reason, studying the effects on biomolecules of extremophilic microorganisms through astrobiological ground-based space simulation experiments is significant to support the interpretation of the data that will be gained and collected during the ongoing and future space exploration missions. Here, the stability of the biomolecules of the ryptoendolithic black fungus Cryomyces antarcticus, grown on two Martian regolith analogues and on Antarctic sandstone, were analysed through a metabolomic approach, after its exposure to Science Verification Tests (SVTs) performed in the frame of the European Space Agency (ESA) Biology and Mars Experiment (BIOMEX) project. These tests are building a set of ground-based experiments performed before the space exposure aboard the International Space Station (ISS). The analysis aimed to investigate the effects of different mineral mixtures on fungal colonies and the stability of the biomolecules synthetised by the fungus under simulated Martian and space conditions. The identification of a specific group of molecules showing good stability after the treatments allow the creation of a molecular database that should support the analysis of future data sets that will be collected in the ongoing and next space exploration missions

Investigation of fungal biomolecules after Low Earth Orbit exposure: a testbed for the next Moon missions

The Moon is characterized by extremely harsh conditions due to ultraviolet irradiation, wide temperature extremes, vacuum resulting from the absence of an atmosphere and high ionizing radiation. Therefore, its surface may provide a unique platform to investigate the effects of such conditions. For lunar exploration with the Lunar Gateway platform, exposure experiments in Low Earth Orbit are useful testbeds to prepare for lunar space experiments and to understand how and if potential biomarkers are influenced by extra-terrestrial conditions. During the BIOMEX (BIOlogy and Mars EXperiment) project, dried colonies of the fungus Cryomyces antarcticus grown on Lunar Regolith Analogue (LRA) were exposed to space conditions for 16 months aboard the EXPOSE-R2 payload outside the International Space Station. In this study, we investigated the stability/degradation of fungal biomarkers in LRA after exposure to (i) simulated space and (ii) real space conditions, using Raman spectroscopy, gas chromatography–mass spectrometry and DNA amplification. The results demonstrated that fungal biomarkers were detectable after 16 months of real space exposure. This work will contribute to the interpretation of data from future biological experiments in the Cislunar orbit with the Lunar Gateway platform and/or on the lunar surface, in preparation for the next step of human exploration

Biological Contamination Prevention for Outer Solar System Moons of Astrobiological Interest: What Do We Need to Know?

To ensure that scientific investments in space exploration are not compromised by terrestrial contamination of celestial bodies, special care needs to be taken to preserve planetary conditions for future astrobiological exploration. Significant effort has been made and is being taken to address planetary protection in the context of inner Solar System exploration. In particular for missions to Mars, detailed internationally accepted guidelines have been established. For missions to the icy moons in the outer Solar System, Europa and Enceladus, the planetary protection requirements are so far based on a probabilistic approach and a conservative estimate of poorly known parameters. One objective of the European Commission-funded project, Planetary Protection of Outer Solar System, was to assess the existing planetary protection approach, to identify inherent knowledge gaps, and to recommend scientific investigations necessary to update the requirements for missions to the icy moons

Experimental and Simulation Efforts in the Astrobiological Exploration of Exooceans

The icy satellites of Jupiter and Saturn are perhaps the most promising places in the Solar System regarding habitability. However, the potential habitable environments are hidden underneath km-thick ice shells. The discovery of Enceladus’ plume by the Cassini mission has provided vital clues in our understanding of the processes occurring within the interior of exooceans. To interpret these data and to help configure instruments for future missions, controlled laboratory experiments and simulations are needed. This review aims to bring together studies and experimental designs from various scientific fields currently investigating the icy moons, including planetary sciences, chemistry, (micro-)biology, geology, glaciology, etc. This chapter provides an overview of successful in situ, in silico, and in vitro experiments, which explore different regions of interest on icy moons, i.e. a potential plume, surface, icy shell, water and brines, hydrothermal vents, and the rocky core