Future Life Detection Missions Research Articles

Radiolytic Effects on Biological and Abiotic Amino Acids in Shallow Subsurface Ices on Europa and Enceladus.

Europa and Enceladus are key targets to search for evidence of life in our solar system. However, the surface and shallow subsurface of both airless icy moons are constantly bombarded by ionizing radiation that could degrade chemical biosignatures. Therefore, sampling of icy surfaces in future life detection missions to Europa and Enceladus requires a clear understanding of the necessary ice depth where unaltered organic biomolecules might be present. We conducted radiolysis experiments by exposing individual amino acids in ices and amino acids from dead microorganisms in ices to gamma radiation to simulate conditions on these icy worlds. In the pure amino acid samples, glycine did not show a detectable decrease in abundance, whereas the abundance of isovaline decreased by 40% after 4 MGy of exposure. Amino acids in dead Escherichia coli (E. coli) organic matter exhibited a gradual decline in abundances with the increase of exposure dosage, although at much slower rates than individual amino acids. The majority of amino acids in dead A. woodii samples demonstrated a step function decline as opposed to a gradual decline. After the initial drop in abundance with 1 MGy of exposure, those amino acids did not display further decreases in abundance after exposure up to 4 MGy. New radiolysis constants for isolated amino acids and amino acids in dead E. coli material for Europa/Enceladus-like conditions have been derived. Slow rates of amino acid destruction in biological samples under Europa and Enceladus-like surface conditions bolster the case for future life detection measurements by Europa and Enceladus lander missions. Based on our measurements, the "safe" sampling depth on Europa is ∼20 cm at high latitudes of the trailing hemisphere in the area of little impact gardening. Subsurface sampling is not required for the detection of amino acids on Enceladus-these molecules will survive radiolysis at any location on the Enceladus surface. If the stability of amino acids observed in A. woodii organic materials is confirmed in other microorganisms, then the survival of amino acids from a potential biosphere in Europa ice would be significantly increased.

Detectability of Surface Biosignatures for Directly Imaged Rocky Exoplanets.

Modeling the detection of life has never been more opportune. With next-generation space telescopes, such as the currently developing Habitable Worlds Observatory (HWO) concept, we will begin to characterize rocky exoplanets potentially similar to Earth. However, few realistic planetary spectra containing surface biosignatures have been paired with direct imaging telescope instrument models. Therefore, we use a HWO instrument noise model to assess the detection of surface biosignatures affiliated with oxygenic, anoxygenic, and nonphotosynthetic extremophiles. We pair the HWO telescope model to a one-dimensional radiative transfer model to estimate the required exposure times necessary for detecting each biosignature on planets with global microbial coverage and varying atmospheric water vapor concentrations. For modeled planets with 0-50% cloud coverage, we determine pigments and the red edge could be detected within 1000 hr (100 hr) at distances within 15 pc (11 pc). However, tighter telescope inner working angles (2.5 λ/D) would allow surface biosignature detection at further distances. Anoxygenic photosynthetic biosignatures could also be more easily detectable than nonphotosynthetic pigments and the photosynthetic red edge when compared against a false positive iron oxide slope. Future life detection missions should evaluate the influence of false positives on the detection of multiple surface biosignatures.

A Review on Hypothesized Metabolic Pathways on Europa and Enceladus: Space-Flight Detection Considerations.

Enceladus and Europa, icy moons of Saturn and Jupiter, respectively, are believed to be habitable with liquid water oceans and therefore are of interest for future life detection missions and mission concepts. With the limited data from missions to these moons, many studies have sought to better constrain these conditions. With these constraints, researchers have, based on modeling and experimental studies, hypothesized a number of possible metabolisms that could exist on Europa and Enceladus if these worlds host life. The most often hypothesized metabolisms are methanogenesis for Enceladus and methane oxidation/sulfate reduction on Europa. Here, we outline, review, and compare the best estimated conditions of each moon's ocean. We then discuss the hypothetical metabolisms that have been suggested to be present on these moons, based on laboratory studies and Earth analogs. We also detail different detection methods that could be used to detect these hypothetical metabolic reactions and make recommendations for future research and considerations for future missions.

Habitability and Biosignature Formation in Simulated Martian Aqueous Environments.

Water present on early Mars is often assumed to have been habitable. In this study, experiments were performed to investigate the habitability of well-defined putative martian fluids and to identify the accompanying potential formation of biosignatures. Simulated martian environments were developed by combining martian fluid and regolith simulants based on the chemistry of the Rocknest sand shadow at Gale Crater. The simulated chemical environment was inoculated with terrestrial anoxic sediment from the Pyefleet mudflats (United Kingdom). These enrichments were cultured for 28 days and subsequently subcultured seven times to ensure that the microbial community was solely grown on the defined, simulated chemistry. The impact of the simulated chemistries on the microbial community was assessed by cell counts and sequencing of 16S rRNA gene profiles. Associated changes to the fluid and precipitate chemistries were established by using ICP-OES, IC, FTIR, and NIR. The fluids were confirmed as habitable, with the enriched microbial community showing a reduction in abundance and diversity over multiple subcultures relating to the selection of specific metabolic groups. The final community comprised sulfate-reducing, acetogenic, and other anaerobic and fermentative bacteria. Geochemical characterization and modeling of the simulant and fluid chemistries identified clear differences between the biotic and abiotic experiments. These differences included the elimination of sulfur owing to the presence of sulfate-reducing bacteria and more general changes in pH associated with actively respiring cells that impacted the mineral assemblages formed. This study confirmed that a system simulating the fluid chemistry of Gale Crater could support a microbial community and that variation in chemistries under biotic and abiotic conditions can be used to inform future life-detection missions.

Mars-like UV Flux and Ionizing Radiation Differently Affect Biomarker Detectability in the Desert Cyanobacterium Chroococcidiopsis as Revealed by the Life Detector Chip Antibody Microarray.

The effect of a Mars-like UV flux and γ-radiation on the detectability of biomarkers in dried cells of Chroococcidiopsis sp. CCMEE 029 was investigated using a fluorescence sandwich microarray immunoassay. The production of anti-Chroococcidiopsis antibodies allowed the immunoidentification of a reduced, though still detectable, signal in dried cells mixed with phyllosilicatic and sulfatic Mars regolith simulants after exposure to 6.8 × 105 kJ/m2 of a Mars-like UV flux. No signal was detected in dried cells that were not mixed with minerals after 1.4 × 105 kJ/m2. For γ-radiation (60Co), no detectable variations of the fluorescence signal occurred in dried cells exposed to 113 kGy compared to non-irradiated dried cells. Our results suggest that immunoassay-based techniques could be used to detect life tracers eventually present in the martian subsurface in freshly excavated materials only if shielded from solar UV. The high structural integrity of biomarkers irradiated with γ-radiation that mimics a dose accumulated in 13 Myr at 2 m depth from the martian surface has implications for the potential detectability of similar organic molecules/compounds by future life-detection missions such as the ExoMars Rosalind Franklin rover.

Directional Aspects of Vegetation Linear and Circular Polarization Biosignatures.

Homochirality is a generic and unique property of all biochemical life and it is considered a universal and agnostic biosignature. Upon interaction with unpolarized light, homochirality induces fractional circular polarization in the light that is scattered from it, which can be sensed remotely. As such, it can be a prime candidate biosignature in the context of future life detection missions and observatories. The linear polarizance of vegetation is also sometimes envisaged as a biosignature, although it does not share the same molecular origin as circular polarization. It is known that linear polarization of surfaces is strongly dependent on the phase angle. The relationship between the phase angle and circular polarization stemming from macromolecular assemblies such as in vegetation, however, remained unclear. In this study, using the average of 27 different species, we demonstrate that the circular polarization-phase angle dependency of vegetation induces relatively small changes in spectral shape and mostly affects the signal magnitude. With these results, we underline the use of circular spectropolarimetry as a promising agnostic biosignature complementary to the use of linear spectropolarimetry and scalar reflectance.

Contamination analysis of Arctic ice samples as planetary field analogs and implications for future life-detection missions to Europa and Enceladus

Missions to detect extraterrestrial life are being designed to visit Europa and Enceladus in the next decades. The contact between the mission payload and the habitable subsurface of these satellites involves significant risk of forward contamination. The standardization of protocols to decontaminate ice cores from planetary field analogs of icy moons, and monitor the contamination in downstream analysis, has a direct application for developing clean approaches crucial to life detection missions in these satellites. Here we developed a comprehensive protocol that can be used to monitor and minimize the contamination of Arctic ice cores in processing and downstream analysis. We physically removed the exterior layers of ice cores to minimize bioburden from sampling. To monitor contamination, we constructed artificial controls and applied culture-dependent and culture-independent techniques such as 16S rRNA amplicon sequencing. We identified 13 bacterial contaminants, including a radioresistant species. This protocol decreases the contamination risk, provides quantitative and qualitative information about contamination agents, and allows validation of the results obtained. This study highlights the importance of decreasing and evaluating prokaryotic contamination in the processing of polar ice cores, including in their use as analogs of Europa and Enceladus.

From Microorganisms to Biosignatures: Subcritical Water Extraction as a Sample Preparation Technique for Future Life Detection Missions

AbstractAmino acids are important targets in the search for life beyond Earth. The types of amino acids, their relative abundances, and enantiomeric excess all serve as biosignatures. To detect such biosignatures, identification and quantification of individual amino acids are required. Regardless of the analytical workflow used, it is critical to release amino acids from the sample without altering their molecular distributions. Subcritical water extraction (SCWE) is a promising technique for the release of organics from a variety of samples that has been under development for flight implementation for over 20 years. However, there is a risk of racemization and/or degradation of amino acids during extraction that could alter their distributions. This work demonstrates for the first time that SCWE (200°C for 30 min) can release amino acids from bacterial cells and spores, while maintaining their native distributions and enantiomeric excess.

A Proposed Geobiology-Driven Nomenclature for Astrobiological In Situ Observations and Sample Analyses.

As the exploration of Mars and other worlds for signs of life has increased, the need for a common nomenclature and consensus has become significantly important for proper identification of nonterrestrial/non-Earth biology, biogenic structures, and chemical processes generated from biological processes. The fact that Earth is our single data point for all life, diversity, and evolution means that there is an inherent bias toward life as we know it through our own planet's history. The search for life “as we don't know it” then brings this bias forward to decision-making regarding mission instruments and payloads. Understandably, this leads to several top-level scientific, theoretical, and philosophical questions regarding the definition of life and what it means for future life detection missions. How can we decide on how and where to detect known and unknown signs of life with a single biased data point? What features could act as universal biosignatures that support Darwinian evolution in the geological context of nonterrestrial time lines? The purpose of this article is to generate an improved nomenclature for terrestrial features that have mineral/microbial interactions within structures and to confirm which features can only exist from life (biotic), features that are modified by biological processes (biogenic), features that life does not affect (abiotic), and properties that can exist or not regardless of the presence of biology (abiogenic). These four categories are critical in understanding and deciphering future returned samples from Mars, signs of potential extinct/ancient and extant life on Mars, and in situ analyses from ocean worlds to distinguish and separate what physical structures and chemical patterns are due to life and which are not. Moreover, we discuss hypothetical detection and preservation environments for extant and extinct life, respectively. These proposed environments will take into account independent active and ancient in situ detection prospects by using previous planetary exploration studies and discuss the geobiological implications within an astrobiological context.

Compact Color Biofinder (CoCoBi): Fast, Standoff, Sensitive Detection of Biomolecules and Polyaromatic Hydrocarbons for the Detection of Life.

We have developed a compact instrument called the "COmpact COlor BIofinder", or CoCoBi, for the standoff detection of biological materials and organics with polyaromatic hydrocarbons (PAHs) using a nondestructive approach in a wide area. The CoCoBi system uses a compact solid state, conductively cooled neodymium-doped yttrium aluminum garnet (Nd:YAG) nanosecond pulsed laser capable of simultaneously providing two excitation wavelengths, 355 and 532 nm, and a compact, sensitive-gated color complementary metal-oxide-semiconductor camera detector. The system is compact, portable, and determines the location of biological materials and organics with PAHs in an area 1590 cm2 wide, from a target distance of 3 m through live video using fast fluorescence signals. The CoCoBi system is highly sensitive and capable of detecting a PAH concentration below 1 part per billion from a distance of 1 m. The color images provide the simultaneous detection of various objects in the target area using shades of color and morphological features. We demonstrate that this unique feature successfully detected the biological remains present in a 150-million-year-old fossil buried in a fluorescent clay matrix. The CoCoBi was also successfully field-tested in Hawaiian ocean water during daylight hours for the detection of natural biological materials present in the ocean. The wide-area and video-speed imaging capabilities of CoCoBi for biodetection may be highly useful in future NASA rover-lander life detection missions.

Geomicrobiological Heterogeneity of Lithic Habitats in the Extreme Environment of Antarctic Nunataks: A Potential Early Mars Analog.

Nunataks are permanent ice-free rocky peaks that project above ice caps in polar regions, thus being exposed to extreme climatic conditions throughout the year. They undergo extremely low temperatures and scarcity of liquid water in winter, while receiving high incident and reflected (albedo) UVA-B radiation in summer. Here, we investigate the geomicrobiology of the permanently exposed lithic substrates of nunataks from Livingston Island (South Shetlands, Antarctic Peninsula), with focus on prokaryotic community structure and their main metabolic traits. Contrarily to first hypothesis, an extensive sampling based on different gradients and multianalytical approaches demonstrated significant differences for most geomicrobiological parameters between the bedrock, soil, and loose rock substrates, which overlapped any other regional variation. Brevibacillus genus dominated on bedrock and soil substrates, while loose rocks contained a diverse microbial community, including Actinobacteria, Alphaproteobacteria and abundant Cyanobacteria inhabiting the milder and diverse microhabitats within. Archaea, a domain never described before in similar Antarctic environments, were also consistently found in the three substrates, but being more abundant and potentially more active in soils. Stable isotopic ratios of total carbon (δ 13C) and nitrogen (δ 15N), soluble anions concentrations, and the detection of proteins involved in key metabolisms via the Life Detector Chip (LDChip), suggest that microbial primary production has a pivotal role in nutrient cycling at these exposed areas with limited deposition of nutrients. Detection of stress-resistance proteins, such as molecular chaperons, suggests microbial molecular adaptation mechanisms to cope with these harsh conditions. Since early Mars may have encompassed analogous environmental conditions as the ones found in these Antarctic nunataks, our study also contributes to the understanding of the metabolic features and biomarker profiles of a potential Martian microbiota, as well as the use of LDChip in future life detection missions.

Microbial diversity and activity in Southern California salterns and bitterns: analogues for remnant ocean worlds.

Concurrent osmotic and chaotropic stress make MgCl2 -rich brines extremely inhospitable environments. Understanding the limits of life in these brines is essential to the search for extraterrestrial life on contemporary and relict ocean worlds, like Mars, which could host similar environments. We sequenced environmental 16S rRNA genes and quantified microbial activity across a broad range of salinity and chaotropicity at a Mars-analogue salt harvesting facility in Southern California, where seawater is evaporated in a series of ponds ranging from kosmotropic NaCl brines to highly chaotropic MgCl2 brines. Within NaCl brines, we observed a proliferation of specialized halophilic Euryarchaeota, which corresponded closely with the dominant taxa found in salterns around the world. These communities were characterized by very slow growth rates and high biomass accumulation. As salinity and chaotropicity increased, we found that the MgCl2 -rich brines eventually exceeded the limits of microbial activity. We found evidence that exogenous genetic material is preserved in these chaotropic brines, producing an unexpected increase in diversity in the presumably sterile MgCl2 -saturated brines. Because of their high potential for biomarker preservation, chaotropic brines could therefore serve as repositories of genetic biomarkers from nearby environments (both on Earth and beyond) making them prime targets for future life-detection missions.

Microbial community distribution in variously altered basalts: Insights into astrobiology sample site selection

Terrestrial basalts represent ecological niches with the potential for microbial habitability. However, little is known about how gradients of physical and/or chemical alteration may impact the distribution of microbial biomass. If higher abundances of microbial biomass are associated with identifiable alteration features, i.e. visible from orbit or rover, such features would become an important tool in the selection of astrobiology targets for future life-detection missions. To examine the associations between biomass abundance and basalt alteration processes, phospholipid fatty acid (PLFA) biomarker analysis was used to assess microbial biomass in basalts representative of distinct alteration categories collected from two Mars analog environments: Craters of the Moon National Monument and Preserve (COTM), Idaho and Hawai’i Volcanoes National Park (HVNP), Hawai’i. PLFA concentration was generally highest in field categorized high-temperature alteration (syn-emplacement) samples within COTM, particularly in those that may have also undergone some degree of secondary weathering. Basalts from HVNP showed more variable biomass abundances, with no clear trend between altered and unaltered basalts. HVNP fumarolic deposits (both active and relict fumarole-associated samples) generally had consistently detectable biomass although temperature appeared to play a role in abundance with the highest temperate fumarole exhibiting the lowest PLFA concentration. PLFA profiles generally had high proportions of brPLFA (including biomarkers of iron- and sulfur-reducing bacteria) and indicated that basalts were dominated by heterotrophic organisms, however no clear trend between community biomarker composition and alteration type or extent was identified. Heterogeneity of microbial biomass within terrestrial basalts suggests that identification of distinct alteration features clearly linked to high biomass (biomarkers) is challenging but that altered materials (syn-emplacement and secondary weathering) and fumarolic deposits are likely promising astrobiology targets.

0.25 Ga Salt Deposits Preserve Signatures of Habitable Conditions and Ancient Lipids.

Polygonal features in a ∼250 million-year-old Permian evaporitic deposit were investigated for their geological and organic content to test the hypothesis that they could preserve the signature of ancient habitable conditions and biological activity. Investigations on evaporitic rock were carried out as part of the MIne Analog Research (MINAR) project at Boulby Mine, the United Kingdom. The edges of the polygons have a higher clay content and contain higher abundances of minerals such as quartz and microcline, and clays such as illite and chlorite, compared with the interior of polygons, suggesting that the edges were preferred locations for the accumulation of weathering products during their formation. The mineral content and its strontium isotope ratio suggest that the material is from continental weathering at the borders of the Permian Zechstein Sea. The edges of the polygons contain material with mean δ13C and δ15N values of -20.8 and 5.3, respectively. Lipids, including alkanes and hopanes, were extracted from the interior and edges of the polygons, which are inferred to represent organic material entrained in the evaporites when they were formed. The presence of long-chain alkanes (C20-C35) that lack a carbon preference, low abundances of C23-C29 hopanes, and lack of marine, evaporitic, or thermal maturity indicators show that lipid biomarkers were, at least in part, potentially derived from a continental source and have not undergone significant thermal maturation since deposition. Lipid extractions using weak acids revealed significantly more lipids than those without acid, potentially indicating that encapsulation was not the only type of preservation mechanism occurring in Boulby salts. These data demonstrate the potential for ancient evaporites and their polygons to preserve information on local geological conditions, ancient habitability, and evidence of life. The data show that analogous martian evaporitic deposits are good targets for future life detection missions and the investigation of ancient martian habitability.

The Search for Chiral Asymmetry as a Potential Biosignature in our Solar System.

The search for evidence of extraterrestrial life in our Solar System is currently guided by our understanding of terrestrial biology and its associated biosignatures. The observed homochirality in all life on Earth, that is, the predominance of "left-handed" or l-amino acids and "right-handed" or d-sugars, is a unique property of life that is crucial for molecular recognition, enzymatic function, information storage and structure and is thought to be a prerequisite for the origin or early evolution of life. Therefore, the detection of l- or d-enantiomeric excesses (ee) of chiral amino acids and sugars could be a powerful indicator for extant or extinct life on another world. However, studies of primitive meteorites have revealed they contain extraterrestrial amino acids and sugar acids (aldonic acids) with large enantiomeric excesses of the same chirality as terrestrial biology resulting from nonbiological processes, complicating the use of chiral asymmetry by itself as a definitive biosignature. Here we review our current knowledge of the distributions and enantiomeric and isotopic compositions of amino acids and polyols found in meteorites compared to terrestrial biology and propose a set of criteria for future life detection missions that can be used to help establish the origin of chiral asymmetry.

Alteration of Proteomes in First-Generation Cultures of Bacillus pumilus Spores Exposed to Outer Space.

Bacillus pumilus SAFR-032 was originally isolated from the Jet Propulsion Lab Spacecraft Assembly Facility and thoroughly characterized for its enhanced resistance to UV irradiation and oxidative stress. This unusual resistance of SAFR-032 is of particular concern in the context of planetary protection and calls for development of novel disinfection techniques to prevent extraterrestrial contamination. Previously, spores of SAFR-032 were exposed for 18 months to a variety of space conditions on board the International Space Station to investigate their resistance to Mars-like conditions and space travel. Here, proteomic characterization of vegetative SAFR-032 cells from space-surviving spores is presented in comparison to a ground control. Vegetative cells of the first passage were processed and subjected to quantitative proteomics using tandem mass tags. Approximately 60% of all proteins encoded by SAFR-032 were identified, and 301 proteins were differentially expressed among the strains. We found that proteins predicted to be involved in carbohydrate transport/metabolism and energy production/conversion had lower abundance than those of the ground control. For three proteins, we showed that the expected metabolic activities were decreased, as expected with direct enzymatic assays. This was consistent with a decrease of ATP production in the space-surviving strains. The same space-surviving strains showed increased abundance of proteins related to survival, growth advantage, and stress response. Such alterations in the proteomes provide insights into possible molecular mechanisms of B. pumilus SAFR-032 to adapt to and resist extreme extraterrestrial environments.IMPORTANCE Spore-forming bacteria are well known for their resistance to harsh environments and are of concern for spreading contamination to extraterrestrial bodies during future life detection missions. Bacillus pumilus has been regularly isolated from spacecraft-associated surfaces and exhibited unusual resistance to ultraviolet light and other sterilization techniques. A better understanding of the mechanisms of microbial survival and enhanced resistance is essential for developing novel disinfection protocols for the purpose of planetary protection. While genomic analyses did not reveal the unique characteristics that explain elevated UV resistance of space-exposed B. pumilus, the proteomics study presented here provided intriguing insight on key metabolic changes. The observed proteomics aberrations reveal a complex biological phenomenon that plays a role in bacterial survival and adaptation under long-term exposure to outer space. This adaptive ability of microorganisms needs to be considered by those tasked with eliminating forward contamination.

Protection of cyanobacterial carotenoids' Raman signatures by Martian mineral analogues after high‐dose gamma irradiation

AbstractThe future robotic exploration missions to Mars—European Space Agency/Roscosmos's ExoMars2020 and National Aeronautics and Space Administration's Mars2020 rovers—will search for signs of extant or extinct life using, among other instruments, Raman spectrometers for the first time. The question remains whether organic biosignatures—such as pigments and cellular components—may be detected by this method. Evaluating their detection limit under simulated extraterrestrial conditions is therefore crucial for the success of future life detection missions. Ionizing radiation can be considered as the most deleterious factor for the long‐term preservation of potential biomarkers on Mars. Here, we report on the preservation potential of Raman signatures in the Antarctic strain CCCryo 231‐06 of the cyanobacterium Nostoc sp. after high doses of gamma irradiation. The carotenoids' signals, a well‐established biosignature model, dominate the Raman spectra at 532‐nm excitation wavelength due to resonance effects. But comparing their distribution and quantifying their preservation are still problematic in natural samples. To standardize the analyses, we successfully applied Raman mapping and signal‐to‐noise ratio masks to quantify the effects of irradiation. The typical in vivo Raman signatures of carotenoids could be detected after exposure to up to 56 kGy with significant deterioration in terms of signal coverage and signal‐to‐noise ratio. But they remained stable even after the highest dose of γ rays (117 kGy) tested in this study for colonies embedded in two different Martian mineral analogues. Data gathered during these ground‐based irradiation experiments contribute to interpret results from space experiments and will guide our search for life on Mars and other bodies of interest.

Electrochemical Detection of the Molecules of Life

Electrochemical sensor devices are of interest in planetary exploration missions where detection of life is a goal. All forms of life on Earth contain cellular machinery that can transform and regulate chemical energy through metabolic pathways. These processes are oxidation-reduction reactions that are performed by four key classes of molecules: flavins, nicotinamides, porphyrins, and quinones (Crawford, 2001). Electrochemical redox may prove a valuable technique for identifying and differentiating these biomolecules and become a useful tool in future life-detecting missions. In this study, we used cyclic voltammetry (CV) and differential pulse voltammetry (DPV) to measure these molecules in a 3D printed flow cell. Electrodes were fabricated from metal wires and interfaced with the flow cell through compression fittings. Glassy carbon (GC) and boron-doped diamond (BDD) electrodes were chosen as working electrodes and used to detect flavin adenine dinucleotide (FAD), riboflavin, plumbagin, anthraquinone, protoporphyrin IX and nicotinamide adenine dinucleotide (NAD). All procedures were performed in a synthetic sea water solution with a Ag/AgCl reference electrode and a platinum wire counter electrode. Limits of detection down to 10 nM were achieved by the GCE. The wide potential window of BDD proved successful in the detection all four classes of redox-active molecules in complex mixtures. Introduction of electrodes into the 3D-printed flow-cell resulted in reliable CV and DPV data, and present this electrode-integrated system as a robust device suitable for future life-detection missions. Crawford, R. (2001). In Search of the Molecules of Life. Icarus, 154(2), 531–539. https://doi.org/10.1006/icar.2001.6714

Biological Characterization of Microenvironments in a Hypersaline Cold Spring Mars Analog.

While many habitable niches on Earth are characterized by permanently cold conditions, little is known about the spatial structure of seasonal communities and the importance of substrate-cell associations in terrestrial cyroenvironments. Here we use the 16S rRNA gene as a marker for genetic diversity to compare two visually distinct but spatially integrated surface microbial mats on Axel Heiberg Island, Canadian high arctic, proximal to a perennial saline spring. This is the first study to describe the bacterial diversity in microbial mats on Axel Heiberg Island. The hypersaline springs on Axel Heiberg represent a unique analog to putative subsurface aquifers on Mars. The Martian subsurface represents the longest-lived potentially habitable environment on Mars and a better understanding of the microbial communities on Earth that thrive in analog conditions will help direct future life detection missions. The microbial mats sampled on Axel Heiberg are only visible during the summer months in seasonal flood plains formed by melt water and run-off from the proximal spring. Targeted-amplicon sequencing revealed that not only does the bacterial composition of the two mat communities differ substantially from the sediment community of the proximal cold spring, but that the mat communities are distinct from any other microbial community in proximity to the Arctic springs studied to date. All samples are dominated by Gammaproteobacteria: Thiotichales is dominant within the spring samples while Alteromonadales comprises a significant component of the mat communities. The two mat samples differ in their Thiotichales:Alteromonadales ratio and contribution of Bacteroidetes to overall diversity. The red mats have a greater proportion of Alteromonadales and Bacteroidetes reads. The distinct bacterial composition of the mat bacterial communities suggests that the spring communities are not sourced from the surface, and that seasonal melt events create ephemerally habitable niches with distinct microbial communities in the Canadian high arctic. The finding that these surficial complex microbial communities exist in close proximity to perennial springs demonstrates the existence of a transiently habitable niche in an important Mars analog site.

Determination of Geochemical Bio-Signatures in Mars-Like Basaltic Environments.

Bio-signatures play a central role in determining whether life existed on early Mars. Using a terrestrial basalt as a compositional analog for the martian surface, we applied a combination of experimental microbiology and thermochemical modeling techniques to identify potential geochemical bio-signatures for life on early Mars. Laboratory experiments were used to determine the short-term effects of biota on the dissolution of terrestrial basalt, and the formation of secondary alteration minerals. The chemoorganoheterotrophic bacterium, Burkholderia sp. strain B_33, was grown in a minimal growth medium with and without terrestrial basalt as the sole nutrient source. No growth was detected in the absence of the basalt. In the presence of basalt, during exponential growth, the pH decreased rapidly from pH 7.0 to 3.6 and then gradually increased to a steady-state of equilibrium of between 6.8 and 7.1. Microbial growth coincided with an increase in key elements in the growth medium (Si, K, Ca, Mg, and Fe). Experimental results were compared with theoretical thermochemical modeling to predict growth of secondary alteration minerals, which can be used as bio-signatures, over a geological timescale. We thermochemically modeled the dissolution of the basalt (in the absence of biota) in very dilute brine at 25°C, 1 bar; the pH was buffered by the mineral dissolution and precipitation reactions. Preliminary results suggested that at the water to rock ratio of 1 × 107, zeolite, hematite, chlorite, kaolinite, and apatite formed abiotically. The biotic weathering processes were modeled by varying the pH conditions within the model to adjust for biologic influence. The results suggested that, for a basaltic system, the microbially-mediated dissolution of basalt would result in “simpler” secondary alteration, consisting of Fe-hydroxide and kaolinite, under conditions where the abiotic system would also form chlorite. The results from this study demonstrate that, by using laboratory-based experiments and thermochemical modeling, it is possible to identify secondary alteration minerals that could potentially be used to distinguish between abiotic and biotic weathering processes on early Mars. This work will contribute to the interpretation of data from past, present, and future life detection missions to Mars.