Natural microbial populations in a water-based biowaste management system for space life support

AbstractThe reutilization of wastewater is a key issue with regard to long-term space missions and planetary habitation. This study reports the design, test runs and microbiological analyses of a fixed bed biofiltration system which applies pumice grain (16–25 mm grain size, 90 m2/m3 active surface) as matrix and calcium carbonate as buffer. For activation, the pumice was inoculated with garden soil known to contain a diverse community of microorganisms, thus enabling the filtration system to potentially degrade all kinds of organic matter. Current experiments over 194 days with diluted synthetic urine (7% and 20%) showed that the 7% filter units produced nitrate slowly but steadily (max. 2191 mg NO3–N/day). In the 20% units nitrate production was slower and less stable (max. 1411 mg NO3–N/day). 84% and 76% of the contained nitrogen was converted into nitrate. The low conversion rate is assumed to be due to the high flow rate, which keeps the biofilm on the pumice thin. At the same time the thin biofilm seems to prevent the activity of denitrifiers implicating the existence of a trade off between rate and the amount of nitrogen loss. Microbiological analyses identified a comparatively low number of species (26 in the filter material, 12 in the filtrate) indicating that urine serves as a strongly selective medium and filter units for the degradation of mixed feedstock have to be pre-conditioned on the intended substrates from the beginning

Limits of life and the habitability of Mars: The ESA space experiment BIOMEX on the ISS

BIOMEX (BIOlogy and Mars EXperiment) is an ESA/Roscosmos space exposure experiment housed within the exposure facility EXPOSE-R2 outside the Zvezda module on the International Space Station (ISS). The design of the multiuser facility supports—among others—the BIOMEX investigations into the stability and level of degradation of space-exposed biosignatures such as pigments, secondary metabolites, and cell surfaces in contact with a terrestrial and Mars analog mineral environment. In parallel, analysis on the viability of the investigated organisms has provided relevant data for evaluation of the habitability of Mars, for the limits of life, and for the likelihood of an interplanetary transfer of life (theory of lithopanspermia). In this project, lichens, archaea, bacteria, cyanobacteria, snow/permafrost algae, meristematic black fungi, and bryophytes from alpine and polar habitats were embedded, grown, and cultured on a mixture of martian and lunar regolith analogs or other terrestrial minerals. The organisms and regolith analogs and terrestrial mineral mixtures were then exposed to space and to simulated Mars-like conditions by way of the EXPOSE-R2 facility. In this special issue, we present the first set of data obtained in reference to our investigation into the habitability of Mars and limits of life. This project was initiated and implemented by the BIOMEX group, an international and interdisciplinary consortium of 30 institutes in 12 countries on 3 continents. Preflight tests for sample selection, results from ground-based simulation experiments, and the space experiments themselves are presented and include a complete overview of the scientific processes required for this space experiment and postflight analysis. The presented BIOMEX concept could be scaled up to future exposure experiments on the Moon and will serve as a pretest in low Earth orbit

MINERALIZATION AND POTENTIAL FOR FOSSILIZATION OF AN EXTREMOTOLERANT BACTERIUM ISOLATED FROM A PAST MARS ANALOG ENVIRONMENT

Introduction: Several decades dedicated to the study of Mars has enabled scientists to understand that, during its history, environmental conditions on early Mars strongly contrasted with the present-day conditions, hostile for life. Indeed, previous (Mars Express, Viking…) and more recent (MSL) missions confirmed that liquid water, heat (volcan-ism, hydrothermalism), organic matter, and redox conditions probably occurred on the planet, thus enabling scientists to seriously consider early Mars as being habitable ans suitable for the emergence of Martian life [1]. However, the detection of past life on Mars, if it existed, also requires that biomarkers (i) be preserved over geological time scales and that (ii) they remained detectable. Therefore, as terrestrial analogues for Mars, astrobiologists are addressing questions related to microbial adaptation, lifestyles and survival in extraterrestrial environments [2]. In this context, the European MASE project (Mars Ana-logues for Space Exploration) aims at better understand-ing habitability, microbial lifestyles and biomarker preservation in such environmental analogues. To do this, one of the goals of MASE is to better characterize the evolution and preservation of diverse biomarkers during the microbial fossilization process [3]

Detecting biochemical evidence for life with the signs of life detector (solid) in an anaerobic microorganism under fossilization conditions

The definitive detection of biosignatures in the context of astrobiological missions to Mars is not without difficulty. Could it be possible to detect biomarkers from an extinct form of life in a very ancient material? The traces of some microorganisms can be well preserved thanks to rapid mineralization of the organisms and cementation of the sediments in which they occur [1]. Thus biosignatures could be indicators of either extant or extinct life, the search for which is one of the main objectives of Mars exploration [1]. The central motivation of the MASE project (Mars Analogues for Space Exploration) is to gain knowledge about the habitability of Mars by the study of the adaptation of anaerobic life forms to extreme environments, their environmental context, and the methods used to detect their biosignatures. Within this background a fundamental target of MASE project is to improve and optimize methods for biosignature detection in samples with low biomass from certain Mars analogue sites. In this context we applied antibody multiarray competitive immunoassay to follow the evolution of specific biochemical signatures from a culture under fossilization conditions. An antibody multiarray competitive immunoassay for the simultaneous detection of compounds of a wide range of molecular sizes or whole spores and cells [2] [3] has revealed as suitable option to achieve this MASE purpose. It consists in a rapid strategy to detect a huge set of different epitopes in extracted samples by a sandwich multiarray immunoassay in a slide or LDChip (Life Detector Chip) where huge range of different antibodies are coated. In this report, we present the results from an experiment in which we followed the biochemical signatures from a growing culture of an isolate of Yersinia sp. in fresh media and in a culture growing under fossilization conditions in silica and gypsum. A decrease in the signal of relative fluorescence of antibody-antigen binding (biomarkers detected) is observed when comparing an untreated Yersinia sp. culture and those induced to mineralization at different time points

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

DNA DOUBLE STRAND BREAK REPAIR DURING HEAD-DOWN-TILT BEDREST: AGBRESA MEETS RADIATION

BACKGROUND Radiation and reduced gravity impose a major burden on health and performance during human spaceflight. While radiation increases cancer risk and limits tissue regeneration, reduced gravity predisposes to musculoskeletal and cardiovascular deconditioning. Deconditioning could conceivably limit the recovery from radiation damage. Our aim was to develop a terrestrial ex vivo model that could be utilized to study the effect of simulated reduced gravity using head-down-tilt bed-rest on repair of ionizing-radiation-induced DNA damage

The astrobiological mission EXPOSE-R on board of the International Space Station

EXPOSE-R flew as the second of the European Space Agency (ESA) EXPOSE multi-user facilities on the International Space Station. During the mission on the external URM-D platform of the Zvezda service module, samples of eight international astrobiology experiments selected by ESA and one Russian guest experiment were exposed to low Earth orbit space parameters from March 10th, 2009 to January 21st, 2011. EXPOSE-R accommodated a total of 1220 samples for exposure to selected space conditions and combinations, including space vacuum, temperature cycles through 273K, cosmic radiation, solar electromagnetic radiation at >110, >170 or >200nm at various fluences up to GJm−2. Samples ranged from chemical compounds via unicellular organisms and multicellular mosquito larvae and seeds to passive radiation dosimeters. Additionally, one active radiation measurement instrument was accommodated on EXPOSE-R and commanded from ground in accordance with the facility itself. Data on ultraviolet radiation, cosmic radiation and temperature were measured every 10s and downlinked by telemetry and data carrier every few months. The EXPOSE-R trays and samples returned to Earth on March 9th, 2011 with Shuttle flight, Space Transportation System (STS)-133/ULF 5, Discovery, after successful total mission duration of 27 months in space. The samples were analysed in the individual investigators laboratories. A parallel Mission Ground Reference experiment was performed on ground with a parallel set of hardware and samples under simulated space conditions following to the data transmitted from the flight missio

Taxonomic and functional analyses of intact microbial communities thriving in extreme, astrobiology-relevant, anoxic sites

Background: Extreme terrestrial, analogue environments are widely used models to study the limits of life and to infer habitability of extraterrestrial settings. In contrast to Earth’s ecosystems, potential extraterrestrial biotopes are usually characterized by a lack of oxygen. Methods: In the MASE project (Mars Analogues for Space Exploration), we selected representative anoxic analogue environments (permafrost, salt-mine, acidic lake and river, sulfur springs) for the comprehensive analysis of their microbial communities. We assessed the microbiome profile of intact cells by propidium monoazide-based amplicon and shotgun metagenome sequencing, supplemented with an extensive cultivation effort. Results: The information retrieved from microbiome analyses on the intact microbial community thriving in the MASE sites, together with the isolation of 31 model microorganisms and successful binning of 15 high-quality genomes allowed us to observe principle pathways, which pinpoint specific microbial functions in the MASE sites compared to moderate environments. The microorganisms were characterized by an impressive machinery to withstand physical and chemical pressures. All levels of our analyses revealed the strong and omnipresent dependency of the microbial communities on complex organic matter. Moreover, we identified an extremotolerant cosmopolitan group of 34 poly-extremophiles thriving in all sites. Conclusions: Our results reveal the presence of a core microbiome and microbial taxonomic similarities between saline and acidic anoxic environments. Our work further emphasizes the importance of the environmental, terrestrial parameters for the functionality of a microbial community, but also reveals a high proportion of living microorganisms in extreme environments with a high adaptation potential within habitability borders

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

TOLERANCE OF ARCHAEA AND BACTERIA AGAINST PERCHLORATE AND HYDROGEN PEROXIDE

Due to the ability of (hyper-) thermophilic Bacteria and Archaea to live in extreme habitats on Earth (e.g. boiling acidic springs, black smoker chimneys, hyper-salinic brines) one could suggest, that these organisms can also outlast other harsh conditions, e.g. prevailing in space or on Mars. On Mars the occurrence of different utilizable nutrition components is limited. The Phoenix lander detected significant amounts (0.4 - 0.6 %) of perchlorate ions in Martian soil. Therefore, we examined the ability of the perchlorate metabolizing Archaeon Archaeoglobus fulgidus as well as phylogenetically deep-branching Bacterium Hydrogenothermus marinus to survive and grow in the presence of perchlorate (NaClO4) and hydrogen peroxide (H₂O₂). The investigated microorganisms were able to tolerate high concentrations of NaClO4 without any changes in their growth pattern. After the addition of 280 mM perchlorate H. marinus showed significant changes in cell morphology. This organism is normally growing as single motile short rods; treated with high concentrations of perchlorate long chains were built. A. fulgidus can tolerate concentrations up to 300 mM. On the contrary, both microorganisms were negatively affected in their survival after a treatment with low concentrations (<50 mM) of H₂O₂. In summary (hyper-) thermophiles have so far unknown high tolerances against cell damaging treatments and may serve as model organisms for future space experiments