Life Detection Missions Research Articles

Influence of mineralogy on the preservation of amino acids under simulated Mars conditions

The detection of organic molecules associated with life on Mars is one of the main goals of future life-searching missions such as the ESA-Roscosmos ExoMars and NASA 2020 mission. In this work we studied the preservation of 25 amino acids that were spiked onto the Mars-relevant minerals augite, enstatite, goethite, gypsum, hematite, jarosite, labradorite, montmorillonite, nontronite, olivine and saponite, and on basaltic lava under simulated Mars conditions. Simulations were performed using the Open University Mars Chamber, which mimicked the main aspects of the martian environment, such as temperature, UV radiation and atmospheric pressure. Quantification and enantiomeric separation of the amino acids were performed using gas-chromatography-mass spectrometry (GC–MS). Results show that no amino acids could be detected on the mineral samples spiked with 1 µM amino acid solution (0.1 µmol of amino acid per gram of mineral) subjected to simulation, possibly due to complete degradation of the amino acids and/or low extractability of the amino acids from the minerals. For higher amino acid concentrations, nontronite had the highest preservation rate in the experiments in which 50 µM spiking solution was used (5 µmol/g), while jarosite and gypsum had a higher preservation rate in the experiments in which 25 and 10 µM spiking solutions were used (2.5 and 1 µmol/g), respectively. Overall, the 3 smectite minerals (montmorillonite, saponite, nontronite) and the two sulfates (gypsum, jarosite) preserved the highest amino acid proportions. Our data suggest that clay minerals preserve amino acids due to their high surface areas and small pore sizes, whereas sulfates protect amino acids likely due to their opacity to UV radiation or by partial dissolution and crystallization and trapping of the amino acids. Minerals containing ferrous iron (such as augite, enstatite and basaltic lava) preserved the lowest amount of amino acids, which is explained by iron (II) catalyzed reactions with reactive oxygen species generated under Mars-like conditions. Olivine (forsterite) preserved more amino acids than the other non-clay silicates due to low or absent ferrous iron. Our results show that D- and L-amino acids are degraded at equal rates, and that there is a certain correlation between preservation/degradation of amino acids and their molecular structure: alkyl substitution in the α-carbon seem to contribute towards amino acid stability under UV radiation. These results contribute towards a better selection of sampling sites for the search of biomarkers on future life detection missions on the surface of Mars.

The chances of detecting life on Mars

Missions to Mars progressively reveal the past and present habitability of the red planet. The current priority for Mars science is the recognition of definitive biosignatures related to past or present life. Success of life detection missions requires choices of the best mission design, location on Mars and particular sample to be analyzed. It is essential therefore to incorporate as much information as possible into the mission planning stages to maximize the precious opportunities provided by robotic operation on Mars. Bayesian statistics allow us to accommodate the many unknowns associated with a mission that has yet to take place. We have used Bayesian statistics to reveal that although in situ missions are less complex the overall probabilities of a successful mission to detect biosignatures on Mars are higher for sample return. If a mission has been designed with safe landing and operation as a priority, recognizing and avoiding those samples that do not contain the target biosignature is the most important characteristic, while for a mission where the best possible samples have been targeted the probability that the sample contains the target biosignature and that it can be correctly detected is the most dominant issue. Usefully, Bayesian statistics can be used to evaluate the chances of detecting past or present life for missions to different landing sites on Mars. A comparative assessment of Eberswelde Crater and Gale Crater indicates a higher probability of success for the latter and the probabilities of success are consistently higher for the sample return mission variant. Bayesian statistics, therefore, can inform future Mars mission planning steps to help maximize the possibility of success.

Purification, biochemical characterization, and implications of an alkali-tolerant catalase from the spacecraft-associated and oxidation-resistant Acinetobacter gyllenbergii 2P01AA.

Herein, we report on the purification, characterization, and sequencing of catalase from Acinetobacter gyllenbergii 2P01AA, an extremely oxidation-resistant bacterium that was isolated from the Mars Phoenix spacecraft assembly facility. The Acinetobacter are dominant members of the microbial communities that inhabit spacecraft assembly facilities and consequently may serve as forward contaminants that could impact the integrity of future life-detection missions. Catalase was purified by using a 3-step chromatographic procedure, where mass spectrometry provided respective subunit and intact masses of 57.8 and 234.6 kDa, which were consistent with a small-subunit tetrameric catalase. Kinetics revealed an extreme pH stability with no loss in activity between pH 5 and 11.5 and provided respective kcat/Km and kcat values of ∼10(7) s(-1) M(-1) and 10(6) s(-1), which are among the highest reported for bacterial catalases. The amino acid sequence was deduced by in-depth peptide mapping, and structural homology suggested that the catalases from differing strains of A. gyllenbergii differ only at residues near the subunit interfaces, which may impact catalytic stability. Together, the kinetic, alkali-tolerant, and halotolerant properties of the catalase from A. gyllenbergii 2P01AA are significant, as they are consistent with molecular adaptations toward the alkaline, low-humidity, and potentially oxidizing conditions of spacecraft assembly facilities. Therefore, these results support the hypothesis that the selective pressures of the assembly facilities impact the microbial communities at the molecular level, which may have broad implications for future life-detection missions.

Biota and biomolecules in extreme environments on Earth: implications for life detection on Mars.

The three main requirements for life as we know it are the presence of organic compounds, liquid water, and free energy. Several groups of organic compounds (e.g., amino acids, nucleobases, lipids) occur in all life forms on Earth and are used as diagnostic molecules, i.e., biomarkers, for the characterization of extant or extinct life. Due to their indispensability for life on Earth, these biomarkers are also prime targets in the search for life on Mars. Biomarkers degrade over time; in situ environmental conditions influence the preservation of those molecules. Nonetheless, upon shielding (e.g., by mineral surfaces), particular biomarkers can persist for billions of years, making them of vital importance in answering questions about the origins and limits of life on early Earth and Mars. The search for organic material and biosignatures on Mars is particularly challenging due to the hostile environment and its effect on organic compounds near the surface. In support of life detection on Mars, it is crucial to investigate analogue environments on Earth that resemble best past and present Mars conditions. Terrestrial extreme environments offer a rich source of information allowing us to determine how extreme conditions affect life and molecules associated with it. Extremophilic organisms have adapted to the most stunning conditions on Earth in environments with often unique geological and chemical features. One challenge in detecting biomarkers is to optimize extraction, since organic molecules can be low in abundance and can strongly adsorb to mineral surfaces. Methods and analytical tools in the field of life science are continuously improving. Amplification methods are very useful for the detection of low concentrations of genomic material but most other organic molecules are not prone to amplification methods. Therefore, a great deal depends on the extraction efficiency. The questions “what to look for”, “where to look”, and “how to look for it” require more of our attention to ensure the success of future life detection missions on Mars.

Characterization of hydrogen peroxide-resistant Acinetobacter species isolated during the Mars Phoenix spacecraft assembly.

The microbiological inventory of spacecraft and the associated assembly facility surfaces represent the primary pool of forward contaminants that may impact the integrity of life-detection missions. Herein, we report on the characterization of several strains of hydrogen peroxide-resistant Acinetobacter, which were isolated during the Mars Phoenix lander assembly. All Phoenix-associated Acinetobacter strains possessed very high catalase specific activities, and the specific strain, A. gyllenbergii 2P01AA, displayed a survival against hydrogen peroxide (no loss in 100 mM H2O2 for 1 h) that is perhaps the highest known among Gram-negative and non-spore-forming bacteria. Proteomic characterizations reveal a survival mechanism inclusive of proteins coupled to peroxide degradation (catalase and alkyl hydroperoxide reductase), energy/redox management (dihydrolipoamide dehydrogenase), protein synthesis/folding (EF-G, EF-Ts, peptidyl-tRNA hydrolase, DnaK), membrane functions (OmpA-like protein and ABC transporter-related protein), and nucleotide metabolism (HIT family hydrolase). Together, these survivability and biochemical parameters support the hypothesis that oxidative tolerance and the related biochemical features are the measurable phenotypes or outcomes for microbial survival in the spacecraft assembly facilities, where the low-humidity (desiccation) and clean (low-nutrient) conditions may serve as selective pressures. Hence, the spacecraft-associated Acinetobacter, due to the conferred oxidative tolerances, may ultimately hinder efforts to reduce spacecraft bioburden when using chemical sterilants, thus suggesting that non-spore-forming bacteria may need to be included in the bioburden accounting for future life-detection missions.

Molecular and Phenetic Characterization of the Bacterial Assemblage of Hot Lake, WA, an Environment with High Concentrations of Magnesium Sulfate, and Its Relevance to Mars.

Hot Lake (Oroville, WA) is an athalassohaline epsomite lake that can have precipitating concentrations of MgSO4 salts, mainly epsomite. Little biotic study has been done on epsomite lakes and it was unclear whether microbes isolated from epsomite lakes and their margins would fall within recognized halotolerant genera, common soil genera, or novel phyla. Our initial study cultivated and characterized epsotolerant bacteria from the lake and its margins. Approximately 100 aerobic heterotrophic microbial isolates were obtained by repetitive streak-plating in high-salt media including either 10% NaCl or 2 M MgSO4. The collected isolates were all bacteria, nearly evenly divided between Gram-positive and Gram-negative clades, the most abundant genera being Halomonas, Idiomarina, Marinobacter, Marinococcus, Nesterenkonia, Nocardiopsis, and Planococcus. Bacillus, Corynebacterium, Exiguobacterium, Kocuria, and Staphylococcus also were cultured. This initial study included culture-independent community analysis of direct DNA extracts of lake margin soil using PCR-based clone libraries and 16S rRNA gene phylogeny. Clones assigned Gram-positive bacterial clades (70% of total clones) were dominated by sequences related to uncultured actinobacteria. There were abundant Deltaproteobacteria clones related to bacterial sulfur metabolisms and clones of Legionella and Coxiella. These epsomite lake microbial communities seem to be divided between bacteria primarily associated with hyperhaline environments rich in NaCl and salinotolerant relatives of common soil organisms. Archaea appear to be in low abundance and none were isolated, despite near-saturated salinities. Growth of microbes at very high concentrations of magnesium and other sulfates has relevance to planetary protection and life-detection missions to Mars, where scant liquid water may form as deliquescent brines and appear as eutectic liquids.

Field-Based Raman Spectroscopic Analyses of an Ordovician Stromatolite

Raman spectrometers are being miniaturized for future life-detection missions on Mars. Field-portable Raman spectrometers, which have similar spectral parameters to the instruments being developed for Mars rovers, have been used to examine extant biosignatures, but they have not yet been used to examine ancient biosignatures. Here, a portable Raman spectrometer was used to analyze an Ordovician stromatolite at the outcrop, revealing both its mineralogy and the presence of sp² carbonaceous material. As stromatolites are often used as proof of the presence of life in Archean rocks and are searched for on Mars, the ability to analyze them in the field with no sample preparation has important ramifications for future Mars missions. However, these results also reveal that a 785 nm excitation source, rather than the 532 nm excitation source planned for future missions, might be a better choice in the search for fossil biosignatures.

Insights into the Extremotolerance ofAcinetobacter radioresistens50v1, a Gram-Negative Bacterium Isolated from the Mars Odyssey Spacecraft

The microbiology of the spacecraft assembly process is of paramount importance to planetary exploration, as the biological contamination that can result from remote-enabled spacecraft carries the potential to impact both life-detection experiments and extraterrestrial evolution. Accordingly, insights into the mechanisms and range of extremotolerance of Acinetobacter radioresistens 50v1, a Gram-negative bacterium isolated from the surface of the preflight Mars Odyssey orbiter, were gained by using a combination of microbiological, enzymatic, and proteomic methods. In summary, A. radioresistens 50v1 displayed a remarkable range of survival against hydrogen peroxide and the sequential exposures of desiccation, vapor and plasma phase hydrogen peroxide, and ultraviolet irradiation. The survival is among the highest reported for non-spore-forming and Gram-negative bacteria and is based upon contributions from the enzyme-based degradation of H(2)O(2) (catalase and alkyl hydroperoxide reductase), energy management (ATP synthase and alcohol dehydrogenase), and modulation of the membrane composition. Together, the biochemical and survival features of A. radioresistens 50v1 support a potential persistence on Mars (given an unintended or planned surface landing of the Mars Odyssey orbiter), which in turn may compromise the scientific integrity of future life-detection missions.

Aqueous extracts of a Mars analogue regolith that mimics the Phoenix landing site do not inhibit spore germination or growth of model spacecraft contaminants Bacillus subtilis 168 and Bacillus pumilus SAFR-032

Because Mars is a primary target for life detection and habitability assessment missions, its exploration is also by necessity a Planetary Protection issue. The recent finding of significant levels of perchlorate (ClO4-) in regolith sampled from the Phoenix landing site raises the question of its potential biotoxicity to putative indigenous martian life, microbial forward contaminants from Earth, or future human visitors. To address this issue, an analogue regolith was constructed based on regolith chemistry data from the Phoenix landing site. A Mars Aqueous Regolith Extract (MARE) was prepared from the Phoenix analogue regolith and analyzed by ion chromatography. The MARE contained (mg/L) the cations Na+ (1411±181), Mg2+ (1051±160), Ca2+ (832±125), and K+ (261±29), and the anions SO42-(5911±993), ClO4-(5316±1767), Cl-(171±25) and F− (2.0±0.4). Nitrogen-containing species NO3-(773±113) and NO2-(6.9±2.3) were also present as a result of regolith preparation procedures, but their relevance to Mars is at present unknown. The MARE was tested for potential toxic effects on two model spacecraft contaminants, the spore-forming bacteria Bacillus subtilis strain 168 and Bacillus pumilus strain SAFR-032. In B. subtilis, spore germination and initial vegetative growth (up to ∼5h) was not inhibited in a rich complex medium prepared with the MARE, but growth after 5h was significantly suppressed in medium prepared using the MARE. Both B. subtilis and B. pumilus exhibited significantly higher rates of spore germination and growth in the MARE vs. DW with no additions (likely due to endogenous spore nutrients), but germination and growth was further stimulated by addition of glucose and a combination of buffered inorganic salts (K2HPO4, KH2PO4, (NH4)2SO4, and MgSO4). The data indicate that the aqueous environment in the regolith from the Phoenix landing site containing high levels of perchlorate does not pose a significant barrier to growth of putative forward contaminants such as B. subtilis and B. pumilus under Earth laboratory conditions.

Survival ofBacillus pumilusSpores for a Prolonged Period of Time in Real Space Conditions

To prevent forward contamination and maintain the scientific integrity of future life-detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated surfaces, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UV radiation and peroxide treatment. Subsequently, B. pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions via the European Technology Exposure Facility (EuTEF). After 18 months of exposure in the EXPOSE facility of the European Space Agency (ESA) on EuTEF under dark space conditions, SAFR-032 spores showed 10-40% survivability, whereas a survival rate of 85-100% was observed when these spores were kept aboard the ISS under dark simulated martian atmospheric conditions. In contrast, when UV (>110 nm) was applied on SAFR-032 spores for the same time period and under the same conditions used in EXPOSE, a ∼7-log reduction in viability was observed. A parallel experiment was conducted on Earth with identical samples under simulated space conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (∼3-log reduction in viability for "UV-Mars," and ∼4-log reduction in viability for "UV-Space"). A comparative proteomics analysis indicated that proteins conferring resistant traits (superoxide dismutase) were present in higher concentration in space-exposed spores when compared to controls. Also, the first-generation cells and spores derived from space-exposed samples exhibited elevated UVC resistance when compared with their ground control counterparts. The data generated are important for calculating the probability and mechanisms of microbial survival in space conditions and assessing microbial contaminants as risks for forward contamination and in situ life detection.

Bacterial growth at the high concentrations of magnesium sulfate found in martian soils.

The martian surface environment exhibits extremes of salinity, temperature, desiccation, and radiation that would make it difficult for terrestrial microbes to survive. Recent evidence suggests that martian soils contain high concentrations of MgSO₄ minerals. Through warming of the soils, meltwater derived from subterranean ice-rich regolith may exist for an extended period of time and thus allow the propagation of terrestrial microbes and create significant bioburden at the near surface of Mars. The current report demonstrates that halotolerant bacteria from the Great Salt Plains (GSP) of Oklahoma are capable of growing at high concentrations of MgSO₄ in the form of 2 M solutions of epsomite. The epsotolerance of isolates in the GSP bacterial collection was determined, with 35% growing at 2 M MgSO₄. There was a complex physiological response to mixtures of MgSO₄ and NaCl coupled with other environmental stressors. Growth also was measured at 1 M concentrations of other magnesium and sulfate salts. The complex responses may be partially explained by the pattern of chaotropicity observed for high-salt solutions as measured by agar gelation temperature. Select isolates could grow at the high salt concentrations and low temperatures found on Mars. Survival during repetitive freeze-thaw or drying-rewetting cycles was used as other measures of potential success on the martian surface. Our results indicate that terrestrial microbes might survive under the high-salt, low-temperature, anaerobic conditions on Mars and present significant potential for forward contamination. Stringent planetary protection requirements are needed for future life-detection missions to Mars.

Spectrofluorometric analysis of amino acid mixtures: Implications for future space missions

Efficient detection of organic molecules is fundamental for the success of future life detection missions. Spectrofluorometric analysis is one of the many techniques that may be used to detect organic molecules in extraterrestrial settings. A particularly important class of organic molecules to target is the amino acids on which all terrestrial life depends. This study aims to identify the optimum fluorescence excitation and emission wavelengths for 17 amino acid standards to examine the effects of amino acid concentrations, mixtures and fluorescence quenching. The results and interpretations can guide the design and operation of life detection protocols on future space missions.

Fluorescence spectroscopy for the detection of life in the Salten Skov Mars regolith analogue

We have performed fluorescence spectroscopic analyses of the amino acid content of the Mars soil analog Salten Skov and of an amino acid standard mixture with the same amino acid abundance and distribution as the soil. This is the first study that investigates the total amino acid abundance of a Mars soil analog with a spectrofluorometer and correlates it with the results obtained from other analytical techniques. We discuss the variations between the fluorescence spectra of the Salten Skov soil sample and the amino acid standard mixture, and the differences between the Salten Skov hydrolyzed and non-hydrolyzed sample fractions. We also discuss the advantages and limitations of fluorescence spectroscopy in the context of future life detection missions.

Extraction of amino acids from soils close to the Mars Desert Research Station (MDRS), Utah

AbstractFuture space missions that aim to detect life should search for molecules that are vital to all living organisms. Although the Viking landers did not find any signs of organic molecules on Mars, signatures of past and/or present life may still exist in the Martian regolith. In this paper, we describe amino acid analyses performed in several Martian analogue soil samples collected close to the Mars Desert Research Station (MDRS), Utah, during the International Lunar Exploration Working Group (ILEWG) EuroGeoMars campaign in February 2009. The Utah desert around Hanksville is characterized as shale desert and is cold and arid with an average annual temperature of 12°C. It is subjected to wind erosion and was shaped by fluvial erosion. The data show large differences in the total amino acid abundances between all the collected soil samples, with values ranging from non-detectable to 100 000 parts per billion (ppb). These results are explained in the context of mineralogical differences (namely different clay content) among the soil samples. The data have implications for future life-detection missions and the target mineralogy that may host biological signatures.

New Priorities in the Robotic Exploration of Mars: The Case for In Situ Search for Extant Life

The search for life on Mars is a priority for NASA’s Science Mission Directorate, a pivotal question of the Astrobiology Program, and the ultimate goal of the Mars Exploration Program (MEPAG, 2008). Also, assessment of the presence or absence of life on Mars is a prerequisite for human exploration in that it will allow mitigation of potential threats to planetary protection. Nevertheless, the Viking missions remain the first, and only, attempt to search for life on the planet (or anywhere else beyond the confines of Earth). Since Viking, and specially in the last decade, robotic missions to Mars have focused on characterizing the physical, chemical, and geological environment as a necessary step to an efficient and systematic search for life. These missions have provided an unprecedented amount of data, and our knowledge of Mars has grown vastly, with respect to its current conditions and geological evolution. In the wake of 15 years of extraordinary efforts, and at a time when the next Decadal Survey is being drafted with recommendations and plans for the exploration of the Solar System, it would seem that the search for life on Mars would be a high priority. However, based on a preliminary disclosure of the main points included in the Decadal Survey during the Astrobiology Science Conference (AbSciCon, April 26–29, 2010, League City, Texas), in situ life-detection missions to Mars will not be recommended. Instead, a Mars Sample Return mission (2018–2022) will play a preeminent role as the only mission in the next 10 years, excluding the Mars Science Laboratory (2011), with the specific task of searching for evidence of past life on Mars. This programmatic strategy is in line with the general consensus reached after the Viking missions, which was that conditions on, or near, the surface of Mars are prohibitive for microbial activity. This assumption has been reinforced with each follow-up mission and is largely based on the fact that Viking, and more recently Phoenix, failed to detect any organic compounds in the surface soils. The constantly low temperatures and hyper-arid conditions on the surface, together with high doses of radiation and the formation of reactive oxidants in the atmosphere, conspire against an extant martian biota in surface soils. Given these observations, the search for extant life has received little support and has not been considered a programmatic priority, and it is on the verge of being removed altogether from the next Decadal Survey. We argue that the strategy for Mars exploration should center on the search for extant life. By extant life, we mean life that is active today or was active during the recent geological past and is now dormant. As we discuss below, the immediate strategy for Mars exploration cannot focus only on past life based on the results of the Viking missions, particularly given that recent analyses call for a re-evaluation of some of these results. It also cannot be based on the assumption that the surface of Mars is uniformly prohibitive for extant life, since research conducted in the past 30 years in extreme environments on Earth has shown that life is possible under extremes of cold and dryness. For these reasons, we recommend a new strategy for the next decade of robotic investigations on Mars, one in which the in situ search for extant life is the first priority. Given the most updated knowledge we have about Mars’ environmental evolution, we call for a long-term architecture of the Mars Exploration Program that is organized around three main goals in the following order of priority: (1) the search for extant life; (2) the search for past life; and (3) sample return. We argue that this is the most efficient approach by which to address, with a high degree of certainty, the question as to whether life exists on Mars.

Validation of a Nylon-Flocked-Swab Protocol for Efficient Recovery of Bacterial Spores from Smooth and Rough Surfaces

In order to meet planetary-protection requirements, culturable bacterial spore loads are measured representatively for the total microbial contamination of spacecraft. However, the National Aeronautics and Space Administration's (NASA's) cotton swab protocols for spore load determination have not changed for decades. To determine whether a more efficient alternative was available, a novel swab was evaluated for recovery of different Bacillus atrophaeus spore concentrations on stainless steel and other surfaces. Two protocols for the nylon-flocked swab (NFS) were validated and compared to the present NASA standard protocol. The results indicate that the novel swab protocols recover 3- to 4-fold more (45.4% and 49.0% recovery efficiency) B. atrophaeus spores than the NASA standard method (13.2%). Moreover, the nylon-flocked-swab protocols were superior in recovery efficiency for spores of seven different Bacillus species, including Bacillus anthracis Sterne (recovery efficiency, 20%). The recovery efficiencies for B. atrophaeus spores from different surfaces showed a variation from 5.9 to 62.0%, depending on the roughness of the surface analyzed. Direct inoculation of the swab resulted in a recovery rate of about 80%, consistent with the results of scanning electron micrographs that allowed detailed comparisons of the two swab types. The results of this investigation will significantly contribute to the cleanliness control of future life detection missions and will provide significant improvement in detection of B. anthracis contamination for law enforcement and security efforts.

Migrating microbes and planetary protection

Since the dawn of the space age, humans have wrestled with concerns about planetary cross-contamination. Spacecraft launched from Earth can transport hitchhiker microbes to new worlds on both robotic and crewed vehicles. As plans proceed for future life detection and human exploration missions, planetary protection considerations are again uppermost on the agendas of mission planners, and microbiologists have an important role to play.

Comment on “Stereo-Specific Glucose Consumption May Be Used to Distinguish Between Chemical and Biological Reactivity on Mars: A Preliminary Test on Earth”

Sun, H.J., Saccomanno, V, Hedlund, B., and McKay, C.P. (‘‘Stereo-Specific Glucose Consumption May Be Used to Distinguish between Chemical and Biological Reactivity on Mars: A Preliminary Test on Earth’’) is an important and timely paper addressing the recently revived interest in the possibility of extant life on Mars. The paper specifically and succinctly proposes to resolve the 33-year aura of doubt surrounding the Viking Labeled Release (LR) life-detection experiment (Levin and Straat, 1976). Reporting on experiments performed on all three domains of life, archaea, bacteria and eukaryotes, Sun et al. convincingly show that chirality preference distinguishes metabolic consumption of substrates from purely physico-chemical reactions in that the latter have no chirality preference. The Sun group proposes their method to determine whether the Viking LR experiment’s positive response was from living microorganisms (Levin, 1997) or from a physico-chemical agent in the martian soil (Klein, 1999). Sun et al. used wet chemistry to determine microbial uptake of glucose from a racemic nutrient solution to demonstrate biological chiral preference, and a respirometer followed by a gas analyzer for CO2 to demonstrate that the strong oxidant permanganate attacks both the Land Disomers of glucose equally. These complex analytical systems would be difficult to instrument for robotic astrobiological use. They are also orders of magnitude less sensitive than the LR radioisotope technique, which raises the question why the researchers did not use that legacy method. The Viking LR addressed the chirality distinction between living and abiotic reactions, but spacecraft constraints prevented the use of separate instruments for the different chiral isomers. However, those isomeric compounds in the nutrient were included in racemic mixtures to avoid missing an alien life with a chirality different from ours. The Chiral LR miniature instrument was subsequently proposed (Levin, 1987) to identify each specific isomer in the nutrient that obtained a response from the soil. Since the Viking LR produced the response Sun et al. propose to investigate, it would seem important to include those LR chiral substrates, Land Dlactate and Land D-alanine, in any return to Mars for the stated purpose of resolving the origin of the Viking LR signals. Thus, it would have been desirable for the Sun group to have included those isomers. There is a more fundamental problem. The method proposed would fail to detect CH4 from methanogens, should they, as increasingly speculated (Krasnopolsky et al., 2004; Mumma et al., 2009), exist on Mars. On the other hand, the LR experiment would readily detect CH4 emitted by the many methanogens that metabolize organic compounds (Escalante-Semerena and Wolf, 1984), including formate, one of the Viking LR nutrients. Strict lithotrophs could also be detected by using H2O as solvent for the LR nutrients instead of H2O and monitoring for C H4 after it passes through a vapor-impermeable membrane or barrier (to exclude H2O(g) from the detector). In this manner, labeled gas evolved from any labeled substrate applied to the test soil can be detected by the LR instrument, as, for another example, H2S from S-labeled cysteine. No special wet chemistry analytical techniques and their respective instrumentations need be applied. Another difficulty is the selection of glucose as the substrate of choice. Prevailing COSPAR requirements (COSPAR, 2002, 2005) prevent the use of glucose in life-detection missions because it cannot withstand the heat of sterilization prescribed. This position was recently supported for soil samples to be taken beneath the UV depth (the Viking LR samples were taken to a depth of 4 cm) by one of the paper’s authors (McKay, 2009). It was for this reason that glucose, despite its strong metabolic attributes, was not included in the Viking LR. In the final analysis, the paper makes a good case for chiral preference as a means to get a widely acceptable answer to the tantalizing question of what the Viking LR found in the surface soil of Mars and for carrying out future astrobiological investigations of detection and comparative biology between terrestrial life and any alien forms.

New evidence for a magmatic influence on the origin of Valles Marineris, Mars

In this paper, we show that the complex geological evolution of Valles Marineris, Mars, has been highly influenced by the manifestation of magmatism (e.g., possible plume activity). This is based on a diversity of evidence, reported here, for the central part, Melas Chasma, and nearby regions, including uplift, loss of huge volumes of material, flexure, volcanism, and possible hydrothermal and endogenic-induced outflow channel activity. Observations include: (1) the identification of a new > 50 km-diameter caldera/vent-like feature on the southwest flank of Melas, which is spatially associated with a previously identified center of tectonic activity using Viking data; (2) a prominent topographic rise at the central part of Valles Marineris, which includes Melas Chasma, interpreted to mark an uplift, consistent with faults that are radial and concentric about it; (3) HiRISE-identified landforms along the floor of the southeast part of Melas Chasma that are interpreted to reveal a volcanic field; (4) CRISM identification of sulfate-rich outcrops, which could be indicative of hydrothermal deposits; (5) GRS K/Th signature interpreted as water–magma interactions and/or variations in rock composition; and (6) geophysical evidence that may indicate partial compensation of the canyon and/or higher density intrusives beneath it. Long-term magma, tectonic, and water interactions (Late Noachian into the Amazonian), albeit intermittent, point to an elevated life potential, and thus Valles Marineris is considered a prime target for future life detection missions.

Archaeal diversity analysis of spacecraft assembly clean rooms

One of the main tasks of NASA's planetary protection program is to prevent the forward contamination of extraterrestrial environments with Earth life, and in turn preserve other planets and the integrity of future life detection missions. Despite information regarding bacterial diversity in NASA's clean rooms, little is known about the presence of Archaea. Archaeal community analysis of spacecraft-associated surfaces is important, as they are considered by some to represent terrestrial life most capable of surviving on Mars. The first insights into the archaeal diversity of clean rooms where spacecraft assembled are attempted. Nucleic acid sequences clustering with uncultivable Archaea within the Eury- and Crenarchaeota were retrieved from 8 of 26 samples collected from several spacecraft assembly clean rooms. Due to their potential capability to survive and proliferate in Martian conditions, screening for Archaea on spacecraft surfaces and instruments that are associated with future life detection missions may be necessary.