An astrophysical view of Earth-based metabolic biosignature gases

Microbial life on Earth uses a wide range of chemical and energetic resources from diverse habitats. An outcome of this microbial diversity is an extensive and varied list of metabolic byproducts. We review key points of Earth-based microbial metabolism that are useful to the astrophysical search for biosignature gases on exoplanets, including a list of primary and secondary metabolism gas byproducts. Beyond the canonical, unique-to-life biosignature gases on Earth (O(2), O(3), and N(2)O), the list of metabolic byproducts includes gases that might be associated with biosignature gases in appropriate exoplanetary environments. This review aims to serve as a starting point for future astrophysical biosignature gas research.     

Microbial stowaways: inimitable survivors or hopeless pioneers?

Abstract The resiliency of prokaryotic life has provided colonization across the globe and in the recesses of Earth's most extreme environments. Horizontal gene transfer provides access to a global bank of genetic resources that creates diversity and allows real-time adaptive potential to the clonal prokaryotic world. We assess the likelihood that this Earth-based strategy could provide survival and adaptive potential, in the case of microbial stowaways off Earth. Key Words: Bacillus-Horizontal gene transfer-Bacteria-Earth Mars-Evolution. Astrobiology 12, 710-715.     

On biogenicity criteria for endolithic microborings on early Earth and beyond

Micron-sized cavities created by the actions of rock-etching microorganisms known as euendoliths are explored as a biosignature for life on early Earth and perhaps Mars. Rock-dwelling organisms can tolerate extreme environmental stresses and are excellent candidates for the colonization of early Earth and planetary surfaces. Here, we give a brief overview of the fossil record of euendoliths in both sedimentary and volcanic rocks. We then review the current understanding of the controls upon the distribution of euendolithic microborings and use these to propose three lines of approach for testing their biogenicity: first, a geological setting that demonstrates a syngenetic origin for the euendolithic microborings; second, microboring morphologies and distributions that are suggestive of biogenic behavior and distinct from ambient inclusion trails; and third, elemental and isotopic evidence suggestive of biological processing. We use these criteria and the fossil record of terrestrial euendoliths to outline potential environments and techniques to search for endolithic microborings on Mars.     

Microbial survival rates of Escherichia coli and Deinococcus radiodurans under low temperature, low pressure, and UV-Irradiation conditions, and their relevance to possible Martian life

Viability rates were determined for microbial populations of Escherichia coli and Deinococcus radiodurans under the environmental stresses of low temperature (-35 degrees C), low-pressure conditions (83.3 kPa), and ultraviolet (UV) irradiation (37 W/m(2)). During the stress tests the organisms were suspended in saltwater soil and freshwater soil media, at variable burial depths, and in seawater. Microbial populations of both organisms were most susceptible to dehydration stress associated with low-pressure conditions, and to UV irradiation. However, suspension in a liquid water medium and burial at larger depths (5 cm) improved survival rates markedly. Our results indicate that planetary surfaces that possess little to no atmosphere and have low water availability do not constitute a favorable environment for terrestrial microorganisms.     

Extremophiles may be irrelevant to the origin of life

In recent years, Bacteria and Archaea have been discovered living in practically every conceivable terrestrial environment, including some previously thought to be too extreme for survival. Exploration of our solar system has revealed a number of extraterrestrial bodies that harbor environments analogous to many of the terrestrial environments in which extremophiles flourish. The recent discovery of more than 105 extrasolar planets suggests that planetary systems are quite common. These three findings have led some to speculate that life is therefore common in the universe, as life as we know it can seemingly survive almost anywhere there is liquid water. It is suggested here that while environments capable of supporting life may be common, this does not in itself support the notion that life is common in the universe. Given that interplanetary transfer of life may be unlikely, the actual origin of life may require specific environmental and geological conditions that may be much less common than the mere existence of liquid water.     

Reassessing the possibility of life on venus: proposal for an astrobiology mission

With their similar size, chemical composition, and distance from the Sun, Venus and Earth may have shared a similar early history. Though surface conditions on Venus are now too extreme for life as we know it, it likely had abundant water and favorable conditions for life when the Sun was fainter early in the Solar System. Given the persistence of life under stabilizing selection in static environments, it is possible that life could exist in restricted environmental niches, where it may have retreated after conditions on the surface became untenable. High-pressure subsurface habitats with water in the supercritical liquid state could be a potential refugium, as could be the zone of dense cloud cover where thermoacidophilic life might have retreated. Technology based on the Stardust Mission to collect comet particles could readily be adapted for a pass through the appropriate cloud layer for sample collection and return to Earth.     

Survival of rock-colonizing organisms after 1.5 years in outer space

Cryptoendolithic microbial communities and epilithic lichens have been considered as appropriate candidates for the scenario of lithopanspermia, which proposes a natural interplanetary exchange of organisms by means of rocks that have been impact ejected from their planet of origin. So far, the hardiness of these terrestrial organisms in the severe and hostile conditions of space has not been tested over extended periods of time. A first long-term (1.5 years) exposure experiment in space was performed with a variety of rock-colonizing eukaryotic organisms at the International Space Station on board the European EXPOSE-E facility. Organisms were selected that are especially adapted to cope with the environmental extremes of their natural habitats. It was found that some-but not all-of those most robust microbial communities from extremely hostile regions on Earth are also partially resistant to the even more hostile environment of outer space, including high vacuum, temperature fluctuation, the full spectrum of extraterrestrial solar electromagnetic radiation, and cosmic ionizing radiation. Although the reported experimental period of 1.5 years in space is not comparable with the time spans of thousands or millions of years believed to be required for lithopanspermia, our data provide first evidence of the differential hardiness of cryptoendolithic communities in space.     

Biological potential of Martian hydrothermal systems

A source of energy to power metabolism may be a limiting factor in the abundance and spatial distribution of past or extant life on Mars. Although a global average of chemical energy available for microbial metabolism and biomass production on Mars has been estimated previously, issues of how the energy is distributed and which particular environments have the greatest potential to support life remain unresolved. We address these issues using geochemical models to evaluate the amounts of chemical energy available in one potential biological environment, Martian hydrothermal systems. In these models, host rock compositions are based upon the compositions of Martian meteorites, which are reacted at high temperature with one of three groundwater compositions. For each model, the values for Gibbs energy of reactions that are important for terrestrial chemosynthetic organisms and likely representative for putative Martian microbes are calculated. Our results indicate that substantial amounts of chemical energy may be available in these systems, depending most sensitively upon the composition of the host rock. From the standpoint of sources of metabolic energy, it is likely that suitable environments exist to support Martian life.     

The search for life on Europa: limiting environmental factors, potential habitats, and Earth analogues

The putative ocean of Europa has focused considerable attention on the potential habitats for life on Europa. By generally clement Earth standards, these Europan habitats are likely to be extreme environments. The objectives of this paper were to examine: (1) the limits for biological activity on Earth with respect to temperature, salinity, acidity, desiccation, radiation, pressure, and time; (2) potential habitats for life on Europa; and (3) Earth analogues and their limitations for Europa. Based on empirical evidence, the limits for biological activity on Earth are: (1) the temperature range is from 253 to 394 K; (2) the salinity range is a(H2O) = 0.6-1.0; (3) the desiccation range is from 60% to 100% relative humidity; (4) the acidity range is from pH 0 to 13; (5) microbes such as Deinococcus are roughly 4,000 times more resistant to ionizing radiation than humans; (6) the range for hydrostatic pressure is from 0 to 1,100 bars; and (7) the maximum time for organisms to survive in the dormant state may be as long as 250 million years. The potential habitats for life on Europa are the ice layer, the brine ocean, and the seafloor environment. The dual stresses of lethal radiation and low temperatures on or near the icy surface of Europa preclude the possibility of biological activity anywhere near the surface. Only at the base of the ice layer could one expect to find the suitable temperatures and liquid water that are necessary for life. An ice layer turnover time of 10 million years is probably rapid enough for preserving in the surface ice layers dormant life forms originating from the ocean. Model simulations demonstrate that hypothetical oceans could exist on Europa that are too cold for biological activity (T < 253 K). These simulations also demonstrate that salinities are high, which would restrict life to extreme halophiles. An acidic ocean (if present) could also potentially limit life. Pressure, per se, is unlikely to directly limit life on Europa. But indirectly, pressure plays an important role in controlling the chemical environments for life. Deep ocean basins such as the Mariana Trench are good analogues for the cold, high-pressure ocean of Europa. Many of the best terrestrial analogues for potential Europan habitats are in the Arctic and Antarctica. The six factors likely to be most important in defining the environments for life on Europa and the focus for future work are liquid water, energy, nutrients, low temperatures, salinity, and high pressures.     

A perspective on the importance of reproductive mode in astrobiology

Reproduction is a vital characteristic of life, and sex is the most common reproductive mode in the eukaryotic world. Sex and reproduction are not necessarily linked mechanisms: Sexuality without reproduction exists, while several forms of asexual reproduction are known. The occurrence of sexuality itself is paradoxical, as it is very costly in evolutionary terms. Most of the hypotheses (more than 20) attempting to explain the prevalence of sex fall into two categories: Sex either creates good gene combinations for adaptation to environments or eliminates bad gene combinations counteracting the accumulation of mutations. In spite of this apparent wealth of beneficial effects of sex, asexuality is not rare. Most eukaryotic, asexual lineages are short-lived and can only persist through the presence of sexual roots, but at least two animal groups, bdelloid rotifers and darwinulid ostracods, seem to claim the status of ancient asexuals. Research on (a)sexuality is relevant to astrobiology in a number of ways. First, strong relationships between the origin and persistence of life in extreme environments and reproductive mode are known. Second, the "habitability" of nonterrestrial environments to life greatly depends on reproductive mode. Whereas asexuals can do equally well or better in harsh environments, they fail to adapt fast enough to changing abiotic and biotic environments. Third, it has been shown that plants reproduce mainly asexually in space, and sperm production and motility in some vertebrates are hampered. Both findings indicate that extraterrestrial life under conditions different from Earth might be dominated by asexual reproduction. Finally, for exchange of biological material between planets, the choice of reproductive mode will be important.     

Survival of Bacillus pumilus spores 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.     

The microbial habitability of weathered volcanic glass inferred from continuous sensing techniques

Basaltic glasses (hyaloclastite) are a widespread habitat for life in volcanic environments, yet their interior physical conditions are poorly characterized. We investigated the characteristics of exposed weathered basaltic glass from a surface outcrop in Iceland, using microprobes capable of continuous sensing, to determine whether the physical conditions in the rock interior are hospitable to microbial life. The material provided thermal protection from freeze-thaw and rapid temperature fluctuations, similar to data reported for other rock types. Water activity experiments showed that at moisture contents less than 13% wet weight, the glass and its weathering product, palagonite, had a water activity below levels suitable for bacterial growth. In pore spaces, however, these higher moisture conditions might be maintained for many days after a precipitation event. Gas exchange between the rock interior and exterior was rapid (< 10 min) when the rocks were dry, but when saturated with water, equilibration took many hours. During this period, we demonstrated the potential for low oxygen conditions within the rock caused by respiratory stimulation of the heterotrophic community within. These conditions might exist within subglacial environments during the formation of the rocks or in micro-environments in the interior of exposed rocks. The experiments showed that microbial communities at the site studied here could potentially be active for 39% of the year, if the depth of the community within the outcrop maintains a balance between access to liquid water and adequate protection from freezing. In the absence of precipitation, the interior of weathered basaltic glass is an extreme and life-limiting environment for microorganisms on Earth and other planets.     

Assessing the plausibility of life on other worlds

As the field of astrobiology matures and search strategies for life on other worlds are developed, the need to analyze in a systematic way the plausibility for life on other planetary systems becomes increasingly apparent. We propose the adoption of a simple plausibility of life (POL) rating system based on specific criteria. Category I applies to any body shown to have conditions essentially equivalent to those on Earth. Category II applies to bodies for which there is evidence of liquid water and sources of energy and where organic compounds have been detected or can reasonably be inferred (Mars, Europa). Category III applies to worlds where conditions are physically extreme but possibly capable of supporting exotic forms of life unknown on Earth (Titan, Triton). Category IV applies to bodies that could have seen the origin of life prior to the development of conditions so harsh as to make its perseverance at present unlikely but conceivable in isolated habitats (Venus, Io). Category V would be reserved for sites where conditions are so unfavorable for life by any reasonable definition that its origin or persistence there cannot be rated a realistic probability (the Sun, gas giant planets). The proposed system is intended to be generic. It assumes that life is based on polymeric chemistry occurring in a liquid medium with uptake and degradation of energy from the environment. Without any additional specific assumptions about the nature of life, the POL system is universally applicable.     

The simulated silicification of bacteria--new clues to the modes and timing of bacterial preservation and implications for the search for extraterrestrial microfossils

Evidence of microbial life on Earth has been found in siliceous rock formations throughout the geological and fossil record. To understand the mechanisms of silicification and thus improve our search patterns for evidence of fossil microbial life in rocks, a series of controlled laboratory experiments were designed to simulate the silicification of microorganisms. The bacterial strains Pseudomonas fluorescens and Desulphovibrio indonensis were exposed to silicifying media. The experiments were designed to determine how exposure time to silicifying solutions and to silicifying solutions of different Si concentration affect the fossilization of microbial biofilms. The silicified biofilms were analyzed using transmission electron microscopy (TEM) in combination with energy-dispersive spectroscopy. Both bacterial species showed evidence of silicification after 24 h in 1,000 ppm silica solution, although D. indonensis was less prone to silicification. The degree of silicification of individual cells of the same sample varied, though such variations decreased with increasing exposure time. High Si concentration resulted in better preservation of cellular detail; the Si concentration was more important than the duration in Si solution. Even though no evidence of amorphous silica precipitation was observed, bacterial cells became permineralized. High-resolution TEM analysis revealed nanometer-sized crystallites characterized by lattice fringe-spacings that match the (10-11) d-spacing of quartz formed within bacterial cell walls after 1 week in 5,000 ppm silica solution. The mechanisms of silicification under controlled laboratory conditions and the implication for silicification in natural environments are discussed, along with the relevance of our findings in the search for early life on Earth and extraterrestrial life.     

Comparative Metagenomics of Two Microbial Mats at Cuatro Ciénegas Basin II: Community Structure and Composition in Oligotrophic Environments

Abstract Microbial mats are self-sustained, functionally complex ecosystems that make good models for the understanding of past and present microbial ecosystems as well as putative extraterrestrial ecosystems. Ecological theory suggests that the composition of these communities might be affected by nutrient availability and disturbance frequency. We characterized two microbial mats from two contrasting environments in the oligotrophic Cuatro Ciénegas Basin: a permanent green pool and a red desiccation pond. We analyzed their taxonomic structure and composition by means of 16S rRNA clone libraries and metagenomics and inferred their metabolic role by the analysis of functional traits in the most abundant organisms. Both mats showed a high diversity with metabolically diverse members and strongly differed in structure and composition. The green mat had a higher species richness and evenness than the red mat, which was dominated by a lineage of Pseudomonas. Autotrophs were abundant in the green mat, and heterotrophs were abundant in the red mat. When comparing with other mats and stromatolites, we found that taxonomic composition was not shared at species level but at order level, which suggests environmental filtering for phylogenetically conserved functional traits with random selection of particular organisms. The highest diversity and composition similarity was observed among systems from stable environments, which suggests that disturbance regimes might affect diversity more strongly than nutrient availability, since oligotrophy does not appear to prevent the establishment of complex and diverse microbial mat communities. These results are discussed in light of the search for extraterrestrial life. Key Words: Cuatro Ciénegas-Metagenomics-Microbial mats-Oligotrophic-Phosphorus limitation-Stromatolites. Astrobiology 12, 659-673.     

"Hairy blobs:" microbial suspects preserved in modern and ancient extremely acid lake evaporites

"Hairy blobs" are unusual clumps of organic bodies and sulfate crystals that have been found in evaporite minerals grown in acid saline lakes. Here, we document modern hairy blobs in halite and gypsum from 5 modern acid saline lakes in southern Western Australia, and Permian hairy blobs trapped in halite from the mid-Permian Opeche Shale in the subsurface of North Dakota. These are among the first microbial remains described from acid saline lake environments. They give clues about the role of microorganisms in the acidity, geochemistry, and mineralogy of these extreme environments. This study also may add to the inventory of life in extreme environments and help predict possible martian life-forms and the method of preservation.     

Establishment of a rearing system of the extremotolerant tardigrade Ramazzottius varieornatus: a new model animal for astrobiology

Studies on the ability of multicellular organisms to tolerate specific environmental extremes are relatively rare compared to those of unicellular microorganisms in extreme environments. Tardigrades are extremotolerant animals that can enter an ametabolic dry state called anhydrobiosis and have high tolerance to a variety of extreme environmental conditions, particularly while in anhydrobiosis. Although tardigrades have been expected to be a potential model animal for astrobiological studies due to their excellent anhydrobiotic and extremotolerant abilities, few studies of tolerance with cultured tardigrades have been reported, possibly due to the absence of a model species that can be easily maintained under rearing conditions. We report the successful rearing of the herbivorous tardigrade, Ramazzottius varieornatus, by supplying the green alga Chlorella vulgaris as food. The life span was 35 +/- 16.4 d, deposited eggs required 5.7 +/- 1.1 d to hatch, and animals began to deposit eggs 9 d after hatching. The reared individuals of this species had an anhydrobiotic capacity throughout their life cycle in egg, juvenile, and adult stages. Furthermore, the reared adults in an anhydrobiotic state were tolerant of temperatures of 90 degrees C and -196 degrees C, and exposure to 99.8% acetonitrile or irradiation with 4000 Gy (4)He ions. Based on their life history traits and tolerance to extreme stresses, R. varieornatus may be a suitable model for astrobiological studies of multicellular organisms.     

The potential for lithoautotrophic life on Mars: application to shallow interfacial water environments

We developed a numerical model to assess the lithoautotrophic habitability of Mars based on metabolic energy, nutrients, water availability, and temperature. Available metabolic energy and nutrient sources were based on a laboratory-produced Mars-analog inorganic chemistry. For this specific reference chemistry, the most efficient lithoautotrophic microorganisms would use Fe(2+) as a primary metabolic electron donor and NO(3)(-) or gaseous O(2) as a terminal electron acceptor. In a closed model system, biomass production was limited by the electron donor Fe(2+) and metabolically required P, and typically amounted to approximately 800 pg of dry biomass/ml ( approximately 8,500 cells/ml). Continued growth requires propagation of microbes to new fecund environments, delivery of fresh pore fluid, or continued reaction with the host material. Within the shallow cryosphere--where oxygen can be accessed by microbes and microbes can be accessed by exploration-lithoautotrophs can function within as little as three monolayers of interfacial water formed either by adsorption from the atmosphere or in regions of ice stability where temperatures are within some tens of degrees of the ice melting point. For the selected reference host material (shergottite analog) and associated inorganic fluid chemistry, complete local reaction of the host material potentially yields a time-integrated biomass of approximately 0.1 mg of dry biomass/g of host material ( approximately 10(9) cells/g). Biomass could also be sustained where solutes can be delivered by advection (cryosuction) or diffusion in interfacial water; however, both of these processes are relatively inefficient. Lithoautotrophs in near-surface thin films of water, therefore, would optimize their metabolism by deriving energy and nutrients locally. Although the selected chemistry and associated model output indicate that lithoautotrophic microbial biomass could accrue within shallow interfacial water on Mars, it is likely that these organisms would spend long periods in maintenance or survival modes, with instantaneous biomass comparable to or less than that observed in extreme environments on Earth.     

Volcano-ice interaction as a microbial habitat on Earth and Mars

Volcano-ice interaction has been a widespread geological process on Earth that continues to occur to the present day. The interaction between volcanic activity and ice can generate substantial quantities of liquid water, together with steep thermal and geochemical gradients typical of hydrothermal systems. Environments available for microbial colonization within glaciovolcanic systems are wide-ranging and include the basaltic lava edifice, subglacial caldera meltwater lakes, glacier caves, and subsurface hydrothermal systems. There is widespread evidence of putative volcano-ice interaction on Mars throughout its history and at a range of latitudes. Therefore, it is possible that life on Mars may have exploited these habitats, much in the same way as has been observed on Earth. The sedimentary and mineralogical deposits resulting from volcano-ice interaction have the potential to preserve evidence of any indigenous microbial populations. These include j?kulhlaup (subglacial outflow) sedimentary deposits, hydrothermal mineral deposits, basaltic lava flows, and subglacial lacustrine deposits. Here, we briefly review the evidence for volcano-ice interactions on Mars and discuss the geomicrobiology of volcano-ice habitats on Earth. In addition, we explore the potential for the detection of these environments on Mars and any biosignatures these deposits may contain.     

Venus, Mars, and the ices on Mercury and the moon: astrobiological implications and proposed mission designs

Venus and Mars likely had liquid water bodies on their surface early in the Solar System history. The surfaces of Venus and Mars are presently not a suitable habitat for life, but reservoirs of liquid water remain in the atmosphere of Venus and the subsurface of Mars, and with it also the possibility of microbial life. Microbial organisms may have adapted to live in these ecological niches by the evolutionary force of directional selection. Missions to our neighboring planets should therefore be planned to explore these potentially life-containing refuges and return samples for analysis. Sample return missions should also include ice samples from Mercury and the Moon, which may contain information about the biogenic material that catalyzed the early evolution of life on Earth (or elsewhere). To obtain such information, science-driven exploration is necessary through varying degrees of mission operation autonomy. A hierarchical mission design is envisioned that includes spaceborne (orbital), atmosphere (airborne), surface (mobile such as rover and stationary such as lander or sensor), and subsurface (e.g., ground-penetrating radar, drilling, etc.) agents working in concert to allow for sufficient mission safety and redundancy, to perform extensive and challenging reconnaissance, and to lead to a thorough search for evidence of life and habitability.