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.     

Near-infrared detection of potential evidence for microscopic organisms on Europa

The possibility of an ocean within the icy shell of Jupiter's moon Europa has established that world as a primary candidate in the search for extraterrestrial life within our Solar System. This paper evaluates the potential to detect evidence for microbial life by comparing laboratory studies of terrestrial microorganisms with measurements from the Galileo Near Infrared Imaging Spectrometer (NIMS). If the interior of Europa at one time harbored life, some evidence may remain in the surface materials. Examination of laboratory spectra of terrestrial extremophiles measured at cryogenic temperatures reveals distorted, asymmetric nearinfrared absorption features due to water of hydration. The band centers, widths, and shapes of these features closely match those observed in the Europa spectra. These features are strongest in reddish-brown, disrupted terrains such as linea and chaos regions. Narrow spectral features due to amide bonds in the microbe proteins provide a means of constraining the abundances of such materials using the NIMS data. The NIMS data of disrupted terrains exhibit distorted, asymmetric near-infrared absorption features consistent with the presence of water ice, sulfuric acid octahydrate, hydrated salts, and possibly as much as 0.2 mg cm(-3) of carbonaceous material that could be of biological origin. However, inherent noise in the observations and limitations of spectral sampling must be taken into account when discussing these findings.     

Microcolonial fungi: survival potential of terrestrial vegetative structures

So far mainly spores or other "differentiated-for-survival" structures were considered to be resistant against extreme environmental constraints (including extraterrestrial challenges). Microcolonial fungi (MCF) are unique growth structures formed by eukaryotic microorganisms inhabiting rock varnish surfaces in terrestrial deserts. They are here proposed as a new object for exobiological study. Sun-exposed desert rocks provide surface habitats with intense solar radiation, a scarce water supply, drastic changes in temperature, and episodic to sporadic availability of nutrients. These challenging conditions reduce the diversity of life to MCF, whose resistance to desiccation and tolerance for ultraviolet (UV) radiation make them survival specialists. Based upon our studies of MCF, we propose that the following mechanisms are universally employed for survival on rock surfaces: (1) compact tissue-like colony organization formed by thermodynamically optimal round cells embedded in extracellular polymeric substances, (2) the presence of several types of UV-absorbing compounds (melanins and mycosporines) and antioxidants (carotenoids, melanins, and mycosporines) that convey multiple stress resistance to desiccation, temperature, and irradiation changes, and (3) intracellular developmental mechanisms typical for these structures.     

Microbial fuel cells applied to the metabolically based detection of extraterrestrial life

Since the 1970s, when the Viking spacecrafts carried out experiments to detect microbial metabolism on the surface of Mars, the search for nonspecific methods to detect life in situ has been one of the goals of astrobiology. It is usually required that a methodology detect life independently from its composition or form and that the chosen biological signature point to a feature common to all living systems, such as the presence of metabolism. In this paper, we evaluate the use of microbial fuel cells (MFCs) for the detection of microbial life in situ. MFCs are electrochemical devices originally developed as power electrical sources and can be described as fuel cells in which the anode is submerged in a medium that contains microorganisms. These microorganisms, as part of their metabolic process, oxidize organic material, releasing electrons that contribute to the electric current, which is therefore proportional to metabolic and other redox processes. We show that power and current density values measured in MFCs that use microorganism cultures or soil samples in the anode are much larger than those obtained with a medium free of microorganisms or sterilized soil samples, respectively. In particular, we found that this is true for extremophiles, which have been proposed as potential inhabitants of extraterrestrial environments. Therefore, our results show that MFCs have the potential to be used for in situ detection of microbial life.     

The BOSS and BIOMEX space experiments on the EXPOSE-R2 mission: Endurance of the desert cyanobacterium Chroococcidiopsis under simulated space vacuum,Martian atmosphere,UVC radiation and temperature extremes.

The proposed space experiments BOSS (Biofilm Organisms Surfing Space) and BIOMEX (BIOlogy and Mars experiment) will take place on the space exposure facility EXPOSE-R2 on the International Space Station (ISS), which is set to be launched in 2014. In BOSS the hypothesis to be tested is that microorganisms grown as biofilms, hence embedded in self-produced extracellular polymeric substances, are more tolerant to space and Martian conditions compared to their planktonic counterparts. Various microbial biofilms have been developed including those obtained from the cyanobacterium Chroococcidiopsis isolated from hot and cold deserts. The prime objective of BIOMEX is to evaluate to what extent biomolecules are resistant to, and can maintain their stability under, space and Mars-like conditions; therefore a variety of pigments and cell components are under investigation to establish a biosignature data base; e.g. a Raman spectral library to be used for extraterrestrial life biosignatures. The secondary objective of BIOMEX is to investigate the endurance of extremophiles, focusing on their interactions with Lunar and Martian mineral analogues. Ground-based studies are currently being carried out in the framework of EVTs (Experiment Verification Tests) by exposing selected organisms to space and Martian simulations. Results on a desert strain of Chroococcidiopsis obtained from the first set of EVT, e.g. space vacuum, Mars atmosphere, UVC radiation, temperature cycles and extremes, suggested that dried biofilms exhibited an enhanced survival compared to planktonic lifestyle. Moreover the protection provided by a Martian mineral analogue (S-MRS) to the sub-cellular integrities of Chroococcidiopsis against UVC radiation supports the endurance of this cyanobacterium under extraterrestrial conditions and its relevance in the development of life detection strategies.     

Life at the wedge: the activity and diversity of arctic ice wedge microbial communities

The discovery of polygonal terrain on Mars underlain by ice heightens interest in the possibility that this water-bearing habitat may be, or may have been, a suitable habitat for extant life. The possibility is supported by the recurring detection of terrestrial microorganisms in subsurface ice environments, such as ice wedges found beneath tundra polygon features. A characterization of the microbial community of ice wedges from the high Arctic was performed to determine whether this ice environment can sustain actively respiring microorganisms and to assess the ecology of this extreme niche. We found that ice wedge samples contained a relatively abundant number of culturable cells compared to other ice habitats (～10(5) CFU·mL(-1)). Respiration assays in which radio-labeled acetate and in situ measurement of CO(2) flux were used suggested low levels of microbial activity, though more sensitive techniques are required to confirm these findings. Based on 16S rRNA gene pyrosequencing, bacterial and archaeal ice wedge communities appeared to reflect surrounding soil communities. Two Pseudomonas sp. were the most abundant taxa in the ice wedge bacterial library (～50%), while taxa related to ammonia-oxidizing Thaumarchaeota occupied 90% of the archaeal library. The tolerance of a variety of isolates to salinity and temperature revealed characteristics of a psychrotolerant, halotolerant community. Our findings support the hypothesis that ice wedges are capable of sustaining a diverse, plausibly active microbial community. As such, ice wedges, compared to other forms of less habitable ground ice, could serve as a reservoir for life on permanently cold, water-scarce, ice-rich extraterrestrial bodies and are therefore of interest to astrobiologists and ecologists alike. .     

Observations from a 4-year contamination study of a sample depth profile through Martian meteorite Nakhla

Morphological, compositional, and biological evidence indicates the presence of numerous well-developed microbial hyphae structures distributed within four different sample splits of the Nakhla meteorite obtained from the British Museum (allocation BM1913,25). By examining depth profiles of the sample splits over time, morphological changes displayed by the structures were documented, as well as changes in their distribution on the samples, observations that indicate growth, decay, and reproduction of individual microorganisms. Biological staining with DNA-specific molecular dyes followed by epifluorescence microscopy showed that the hyphae structures contain DNA. Our observations demonstrate the potential of microbial interaction with extraterrestrial materials, emphasize the need for rapid investigation of Mars return samples as well as any other returned or impactor-delivered extraterrestrial materials, and suggest the identification of appropriate storage conditions that should be followed immediately after samples retrieved from the field are received by a handling/curation facility. The observations are further relevant in planetary protection considerations as they demonstrate that microorganisms may endure and reproduce in extraterrestrial materials over long (at least 4 years) time spans. The combination of microscopy images coupled with compositional and molecular staining techniques is proposed as a valid method for detection of life forms in martian materials as a first-order assessment. Time-resolved in situ observations further allow observation of possible (bio)dynamics within the system.     

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.     

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 methanogens during desiccation: implications for life on Mars

The relatively recent discoveries that liquid water likely existed on the surface of past Mars and that methane currently exists in the martian atmosphere have fueled the possibility of extant or extinct life on Mars. One possible explanation for the existence of the methane would be the presence of methanogens in the subsurface. Methanogens are microorganisms in the domain Archaea that can metabolize molecular hydrogen as an energy source and carbon dioxide as a carbon source and produce methane. One factor of importance is the arid nature of Mars, at least at the surface. If one is to assume that life exists below the surface, then based on the only example of life that we know, liquid water must be present. Realistically, however, that liquid water may be seasonal just as it is at some locations on our home planet. Here we report on research designed to determine how long certain species of methanogens can survive desiccation on a Mars soil simulant, JSC Mars-1. Methanogenic cells were grown on JSC Mars-1, transferred to a desiccator within a Coy anaerobic environmental chamber, and maintained there for varying time periods. Following removal from the desiccator and rehydration, gas chromatographic measurements of methane indicated survival for varying time periods. Methanosarcina barkeri survived desiccation for 10 days, while Methanobacterium formicicum and Methanothermobacter wolfeii were able to survive for 25 days.     

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 search for extraterrestrial artifacts (SETA)

The rationale for the use of interstellar artifacts by intelligent life in the universe is described. The advantages of using interstellar probes as a means of exploration and communication are presented and shown to be significant enough to counter the time, energy, and technology arguments generally raised against contact via extraterrestrial artifacts. Four classes of artifacts are defined: Those seeking contact, those seeking to avoid contact, those intended to provide a passive technological threshold for detection, and those for which detection is irrelevant. The Search for Extraterrestrial Artifacts (SETA) is based on the latter two classes. Under the assumption that an extraterrestrial probe will be interested in life in our solar system, a near-Earth search space is defined. This search space is accessible to us now with ground and satellite observing facilities. The current observational status of SETA is reviewed and contrasted with the achievable detection limits for the different parts of the search space.     

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.     

Finding extraterrestrial organisms living on thermosynthesis

During thermal cycling, organisms could live on thermosynthesis, a theoretical mechanism applicable to the origin of life and the early evolution of biological energy conversion. All extraterrestrial ice may be a repository for frozen dead or dormant organisms from earlier stages of evolution. In the presence of a thermal gradient within the ice, organisms might still be harvesting energy from thermosynthesis. Possible habitats for thermosynthesizers can be found throughout the Solar System, particularly in the cold traps on Mercury and the Moon, convecting waters on Mars, the oceans on moons in the outer Solar System, and smaller bodies rotating in the sunlight such as cosmic dust, meteorites, asteroids, and comets. A general strategy for detecting thermosynthetic organisms on Earth is offered, and highlights of current and upcoming robotic exploratory missions relevant to the detection of thermosynthesis are reviewed.     

Searching for liquid water in Europa by using surface observatories

Liquid water, as far as we know, is an indispensable ingredient of life. Therefore, locating reservoirs of liquid water in extraterrestrial bodies is a necessary prerequisite to searching for life. Recent geological and geophysical observations from the Galileo spacecraft, though not unambiguous, hint at the possibility of a subsurface ocean in the Jovian moon Europa. After summarizing present evidence for liquid water in Europa, we show that electromagnetic and seismic observations made from as few as two surface observatories comprising a magnetometer and a seismometer offer the best hope of unambiguous characterization of the three-dimensional structure of the ocean and the deeper interior of this icy moon. The observatories would also help us infer the composition of the icy crust and the ocean water.     

Search for extraterrestrial life using chiral molecules: mandelate racemase as a test case

We have investigated an enzymatic racemization reaction as a marker for extraterrestrial life, which resulted in a change in optical rotation of a mandelic acid over time, as measured by polarimetry. Mandelate racemase was active in aqueous buffer in a temperature range between 0 degrees C and 70 degrees C and also in concentrated ammonium salt solutions and water-in-oil microemulsions in a temperature range between -30 degrees C and 60-70 degrees C; however, the enzyme was not active in several organic cryosolvents. Thus, we have demonstrated that concentrated ammonium salt solutions and water-in-oil microemulsions, both of which are able to form on extraterrestrial planets and moons in the presence of liquid water, are suitable media for enzyme reactions at subzero temperatures. Kinetic data for the mandelate racemase reaction obtained by polarimetry, while reproducible and internally consistent, differed significantly from several sets of data obtained previously by other methods such as chromatography and hydrogen-deuterium exchange. However, we conclude that reactions yielding a polarimetric signal, such as the racemizations employed in this work, are suitable mechanisms by which to utilize a change in chirality over time as a tool to detect signs of life.     

Morphological biosignatures and the search for life on Mars

This report provides a rationale for the advances in instrumentation and understanding needed to assess claims of ancient and extraterrestrial life made on the basis of morphological biosignatures. Morphological biosignatures consist of bona fide microbial fossils as well as microbially influenced sedimentary structures. To be recognized as evidence of life, microbial fossils must contain chemical and structural attributes uniquely indicative of microbial cells or cellular or extracellular processes. When combined with various research strategies, high-resolution instruments can reveal such attributes and elucidate how morphological fossils form and become altered, thereby improving the ability to recognize them in the geological record on Earth or other planets. Also, before fossilized microbially influenced sedimentary structures can provide evidence of life, criteria to distinguish their biogenic from non-biogenic attributes must be established. This topic can be advanced by developing process-based models. A database of images and spectroscopic data that distinguish the suite of bona fide morphological biosignatures from their abiotic mimics will avoid detection of false-positives for life. The use of high-resolution imaging and spectroscopic instruments, in conjunction with an improved knowledge base of the attributes that demonstrate life, will maximize our ability to recognize and assess the biogenicity of extraterrestrial and ancient terrestrial life.     

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.     

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.