Astrobiology

Epsomite (MgSO(4).7H(2)O) and hexahydrite (MgSO(4).6H(2)O) are common minerals found in marine evaporite deposits, in saline lakes as precipitates, in weathering zones of coal and metallic deposits, in some soils and their efflorescences, and possibly on the surface of Europa as evaporite deposits. Thermodynamic properties of these two minerals reported in the literature are in poor agreement. In this study, epsomite-hexahydrite equilibria were determined along four humidity-buffer curves at 0.1 MPa and between 25 and 45 degrees C. Results obtained for the reaction epsomite = hexahydrite + H(2)O, as demonstrated by very tight reversals along each humidity buffer, can be represented by ln K(+/- 0.012) = 20.001 - 7182.07/T, where K is the equilibrium constant, and T is temperature in Kelvin. The derived standard Gibbs free energy of reaction is 10.13 +/- 0.07 kJ/mol, which is essentially the same value as that calculated from vapor pressure measurements reported in the literature. However, this value is at least 0.8 kJ/mol lower than those calculated from the data derived mostly from calorimetric measurements.

Abstract We report the first observation of indigenous carbonaceous matter in the martian meteorite Yamato 000593. The carbonaceous phases are heterogeneously distributed within secondary iddingsite alteration veins and present in a range of morphologies including areas composed of carbon-rich spheroidal assemblages encased in multiple layers of iddingsite. We also observed microtubular features emanating from iddingsite veins penetrating into the host olivine comparable in shape to those interpreted to have formed by bioerosion in terrestrial basalts. Key Words: Meteorite-Yamato 000593-Mars-Carbon. Astrobiology 14, 170-181.

Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars.

The multi-user facility EXPOSE-E was designed by the European Space Agency to enable astrobiology research in space (low-Earth orbit). On 7 February 2008, EXPOSE-E was carried to the International Space Station (ISS) on the European Technology Exposure Facility (EuTEF) platform in the cargo bay of Space Shuttle STS-122 Atlantis. The facility was installed at the starboard cone of the Columbus module by extravehicular activity, where it remained in space for 1.5 years. EXPOSE-E was returned to Earth with STS-128 Discovery on 12 September 2009 for subsequent sample analysis. EXPOSE-E provided accommodation in three exposure trays for a variety of astrobiological test samples that were exposed to selected space conditions: either to space vacuum, solar electromagnetic radiation at >110 nm and cosmic radiation (trays 1 and 3) or to simulated martian surface conditions (tray 2). Data on UV radiation, cosmic radiation, and temperature were measured every 10 s and downlinked by telemetry. A parallel mission ground reference (MGR) experiment was performed on ground with a parallel set of hardware and samples under simulated space conditions. EXPOSE-E performed a successful 1.5-year mission in space.

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.

Because of their ubiquity and resistance to spacecraft decontamination, bacterial spores are considered likely potential forward contaminants on robotic missions to Mars. Thus, it is important to understand their global responses to long-term exposure to space or martian environments. As part of the PROTECT experiment, spores of B. subtilis 168 were exposed to real space conditions and to simulated martian conditions for 559 days in low-Earth orbit mounted on the EXPOSE-E exposure platform outside the European Columbus module on the International Space Station. Upon return, spores were germinated, total RNA extracted, fluorescently labeled, and used to probe a custom Bacillus subtilis microarray to identify genes preferentially activated or repressed relative to ground control spores. Increased transcript levels were detected for a number of stress-related regulons responding to DNA damage (SOS response, SPβ prophage induction), protein damage (CtsR/Clp system), oxidative stress (PerR regulon), and cell envelope stress (SigV regulon). Spores exposed to space demonstrated a much broader and more severe stress response than spores exposed to simulated martian conditions. The results are discussed in the context of planetary protection for a hypothetical journey of potential forward contaminant spores from Earth to Mars and their subsequent residence on Mars.

Microfossils preserved in chert from the;1.9 Ga Gunflint Formation (Schreiber Beach, Ontario, Canada) were studied with transmission electron microscopy (TEM) and analytical TEM (ATEM). Our goals were to uncover the style of silicification relative to the distribution of organic matter, and to evaluate the distribution and evolution of organic matter, at submicroscopic spatial scales. Petrographically the microfossils typically display filamentous or coccoidal morphologies, and consist of quartz crystals surrounded by kerogen along grain boundaries. ATEM analysis revealed that quartz associated with kerogen consists of 200-500nm-sized, round crystallites, whereas the chert matrix is comprised of randomly oriented, polygonal microquartz (5-10 microm). Silica spheroids found within some fossils consist of quartz subgrains in an amorphous to poorly crystalline matrix, suggesting that precipitation of opaline silica on organic matter occurred with subsequent but incomplete transformation to quartz. Some coccoidal microfossils surround large euhedral quartz crystals (up to 5 microm in diameter) that appeared to have influenced the distribution of kerogen during crystal growth. These euhedral quartz crystals commonly contain elongated (50-100 nm) iron-rich crystallites. Energy-loss, near-edge structure analysis of kerogen associated with a coccoidal microfossil showed that it is composed of amorphous carbon with no evidence of graphitization. TEM results revealed significant differences in the style of silicification between microbe-shaped microfossils and their surrounding chert matrix, as well as the presence of amorphous kerogen.

The nematode Caenorhabditis elegans, a popular organism for biological studies, is being developed as a model system for space biology. The chemically defined liquid medium, C. elegans Maintenance Medium (CeMM), allows axenic cultivation and automation of experiments that are critical for spaceflight research. To validate CeMM for use during spaceflight, we grew animals using CeMM and standard laboratory conditions onboard STS-107, space shuttle Columbia. Tragically, the Columbia was destroyed while reentering the Earth's atmosphere. During the massive recovery effort, hardware that contained our experiment was found. Live animals were observed in four of the five recovered canisters, which had survived on both types of media. These data demonstrate that CeMM is capable of supporting C. elegans during spaceflight. They also demonstrate that animals can survive a relatively unprotected reentry into the Earth's atmosphere, which has implications with regard to the packaging of living material during space flight, planetary protection, and the interplanetary transfer of life.

Biogenic stromatolites are sources of significant information on the evolution of microbial life. Despite their evolutionary significance, little is known about the mechanisms of osmoadaptation by microorganisms that comprise living stromatolites thriving in hypersaline environments. Osmoadaptive strategies for Halococcus hamelinensis, a novel halophilic archaeon recently isolated from living stromatolites in the hypersaline reaches of Shark Bay, were thus a particular interest in this study. To investigate the possibility of "salt-in-cytoplasm"-associated osmoadaptation for this archaeon, flame photometry studies were performed. From the results, it was evident that this halophilic archaeon did not accumulate intracellular K(+) ions when cells were exposed to either osmotic shock or conditions with gradual increments in salinity. These results were further supported by polymerase chain reaction (PCR) analyses where there was no evidence for the existence of homologous genes to an ATP-driven, high-affinity potassium uptake system in Halococcus hamelinensis. To identify an alternative salt adaptation mechanism associated with accumulation of compatible solutes for this archaeon, (1)H nuclear magnetic resonance (NMR) spectroscopy experiments were carried out. Results indicate that glycine betaine, trehalose, and glutamate are solutes likely to be involved in osmoregulation in this archeaon. Subsequent (1)H NMR analysis of cell extracts from this microorganism grown under various NaCl concentrations revealed that intracellular levels of glycine betaine increased with increasing concentrations of NaCl. This behavior of increasing glycine betaine concentration with increasing external NaCl is consistent with its identity as an osmolyte. In contrast, intracellular levels of trehalose were decreased in high concentrations of NaCl. This provides evidence that compatible solute accumulation appears to be the preferential salt regulation mechanism for this haloarchaeon, in contrast to the salt-in-cytoplasm strategy employed by many other halophilic archaea.

Dry heat microbial reduction is the NASA-approved sterilization method to reduce the microbial bioburden on spaceflight hardware for missions with planetary protection requirements. The method involves heating the spaceflight hardware to temperatures between 104 degrees C and 125 degrees C for up to 50 hours, while controlling the humidity to very low values. Collection of lethality data at temperatures above 125 degrees C and with ambient (uncontrolled) humidity conditions would establish whether any microbial reduction credit can be offered to the flight project for processes that occur at temperatures greater than 125 degrees C. The goal of this research is to determine the survival rates of Bacillus atrophaeus (ATCC 9372) spores subjected to temperatures higher than 125 degrees C under both dry (controlled) and room ambient humidity (36-66% relative humidity) conditions. Spores were deposited inside thin, stainless steel thermal spore exposure vessels (TSEVs) and heated under ambient or controlled humidity conditions from 115 degrees C to 170 degrees C. After the exposures, the TSEVs were cooled rapidly, and the spores were recovered and plated. Survivor ratios, lethality rate constants, and D-values were calculated at each temperature. At 115 degrees C and 125 degrees C, the controlled humidity lethality rate constant was faster than the ambient humidity lethality rate constant. At 135 degrees C, the ambient and controlled humidity lethality rate constants were statistically identical. At 150 degrees C and 170 degrees C, the ambient humidity lethality rate constant was slightly faster than the controlled humidity lethality rate constant. These results provide evidence for possibly modifying the NASA dry heat microbial reduction specification.

Exposing flight hardware to dry heat is a NASA-approved sterilization method for reducing microbial bioburden on spacecraft. The existing NASA specification only allows heating the flight hardware between 104°C and 125°C to reduce the number of viable microbes and bacterial spores. Also, the NASA specifications only allow a four log reduction by dry heat microbial reduction because very heat-resistant spores are presumed to exist in a diverse population (0.1%). The goal of this research was to obtain data at higher temperatures than 125°C for one of the most heat-resistant microorganisms discovered in a spacecraft assembly area. These data support expanding the NASA specifications to temperatures higher than 125°C and relaxing the four log reduction specification. Small stainless steel vessels with spores of the Bacillus strain ATCC 29669 were exposed to constant temperatures between 125°C and 200°C under both dry and ambient room humidity for set time durations. After exposures, the thermal spore exposure vessels were cooled and the remaining spores recovered and plated out. Survivor ratios, lethality rate constants, and D-values were determined at each temperature. The D-values for the spores exposed under dry humidity conditions were always found to be shorter than those under ambient humidity. The temperature dependence of the lethality rate constants was obtained by assuming that they obeyed Arrhenius behavior. The results are compared to those of B. atrophaeus ATCC 9372. In all cases, the D-values of ATCC 29669 are between 20 and 50 times longer than those of B. atrophaeus ATCC 9372.

The bacterial diversity and comparative community structure of a clean room used for assembling the Phoenix spacecraft was characterized throughout the spacecraft assembly process by using 16S rRNA gene cloning/sequencing and DNA microarray (PhyloChip) technologies. Samples were collected from several locations of the clean room at three time points: before Phoenix's arrival (PHX-B), during hardware assembly (PHX-D), and after the spacecraft was removed for launch (PHX-A). Bacterial diversity comprised of all major bacterial phyla of PHX-B was found to be statistically different from PHX-D and PHX-A samples. Due to stringent cleaning and decontamination protocols during assembly, PHX-D bacterial diversity was dramatically reduced when compared to PHX-B and PHX-A samples. Comparative community analysis based on PhyloChip results revealed similar overall trends as were seen in clone libraries, but the high-density phylogenetic microarray detected larger diversity in all sampling events. The decrease in community complexity in PHX-D compared to PHX-B, and the subsequent recurrence of these organisms in PHX-A, speaks to the effectiveness of NASA cleaning protocols. However, the persistence of a subset of bacterial signatures throughout all spacecraft assembly phases underscores the need for continued refinement of sterilization technologies and the implementation of safeguards that monitor and inventory microbial contaminants.

We used Time Domain (1)H Nuclear Magnetic Resonance (NMR) to characterize changes in proton exchange between water and sugar enantiomers at different concentrations of H(2)(17)O (approximately 15-450 mM) and found that dissociation of the (-)-enantiomers of glucose and ribose occurs at significantly higher rates at higher concentrations of H(2)(17)O. The mechanism behind this enantioselective effect is unclear. The hypothesis we propose is that the large magnetic field (B(o) approximately 0.6T) applied during NMR measurements induces electric moments opposite in sign for the D and L-isomers. Because (17)O has a nuclear electric quadrupole moment not = 0, asymmetrically hydrated complexes may form between the B(o)-polarized enantiomers and H(2)(17)O. Either H(2)(17)O is more often hydrating the (+) than the (-)-enantiomers--and consequently pK differences between H(2)(16)O and H(2)(17)O lead to differences in proton exchange between enantiomers and water--or the orientation of H(2)(17)O relative to the B(o)-polarized enantiomers is different, in total or in part, which leads to hydrated complexes with different spatial geometries and different proton exchange properties. This effect is significant for Magneto-Chiral Stereo-Chemistry (MCSC) and astrobiology, and it may help us better understand specific instances of mass independent isotopic fractionation and aid in the development of new technologies for chiral and isotopic separation.

The plausibility that life was imported to Earth from elsewhere can be tested by subjecting life-forms to space travel. Ultraviolet light is the major liability in short-term exposures (Horneck et al., 2001 ), and plant seeds, tardigrades, and lichens-but not microorganisms and their spores-are candidates for long-term survival (Anikeeva et al., 1990 ; Sancho et al., 2007 ; Jönsson et al., 2008 ; de la Torre et al., 2010 ). In the present study, plant seeds germinated after 1.5 years of exposure to solar UV, solar and galactic cosmic radiation, temperature fluctuations, and space vacuum outside the International Space Station. Of the 2100 exposed wild-type Arabidopsis thaliana and Nicotiana tabacum (tobacco) seeds, 23% produced viable plants after return to Earth. Survival was lower in the Arabidopsis Wassilewskija ecotype and in mutants (tt4-8 and fah1-2) lacking UV screens. The highest survival occurred in tobacco (44%). Germination was delayed in seeds shielded from solar light, yet full survival was attained, which indicates that longer space travel would be possible for seeds embedded in an opaque matrix. We conclude that a naked, seed-like entity could have survived exposure to solar UV radiation during a hypothetical transfer from Mars to Earth. Chemical samples of seed flavonoid UV screens were degraded by UV, but their overall capacity to absorb UV was retained. Naked DNA encoding the nptII gene (kanamycin resistance) was also degraded by UV. A fragment, however, was detected by the polymerase chain reaction, and the gene survived in space when protected from UV. Even if seeds do not survive, components (e.g., their DNA) might survive transfer over cosmic distances.

Radiation Risk Radiometer-Dosimeter E (R3DE) served as a device for measuring ionizing and non-ionizing radiation as well as cosmic radiation reaching biological samples located on the EXPOSE platform EXPOSE-E. The duration of the mission was almost 1.5 years (2008-2009). With four channels, R3DE detected the wavelength ranges of photosynthetically active radiation (PAR, 400-700 nm), UVA (315-400 nm), UVB (280-315 nm), and UVC (<280 nm). In addition, the temperature was recorded. Cosmic ionizing radiation was assessed with a 256-channel spectrometer dosimeter (see separate report in this issue). The light and UV sensors of the device were calibrated with spectral measurement data obtained by the Solar Radiation and Climate Experiment (SORCE) satellite as standard. The data were corrected with respect to the cosine error of the diodes. Measurement frequency was 0.1 Hz. Due to errors in data transmission or temporary termination of EXPOSE power, not all data could be acquired. Radiation was not constant during the mission. At regular intervals of about 2 months, low or almost no radiation was encountered. The radiation dose during the mission was 1823.98 MJ m(-2) for PAR, 269.03 MJ m(-2) for UVA, 45.73 MJ m(-2) for UVB, or 18.28 MJ m(-2) for UVC. Registered sunshine duration during the mission was about 152 days (about 27% of mission time).The surface of EXPOSE was most likely turned away from the Sun for considerably longer. R3DE played a crucial role on EXPOSE-EuTEF (EuTEF, European Technology Exposure Facility), because evaluation of the astrobiology experiments depended on reliability of the data collected by the device. Observed effects in the samples were weighted by radiation doses measured by R3DE.

Abstract The aim of this article is to show how, and in which context, astrobiological reasoning was employed before the establishment of astrobiology as a scientific discipline. By way of an example, I will discuss a popular science book published in 1931 by the Hungarian journalist Desiderius Papp. The author claims that this book represents an innovation in astrobiological reasoning, as it draws on contemporary biological research to conduct thought experiments, thereby coming up with concrete forms of possible extraterrestrial life. One of the most interesting of these forms was crystalline life. After a short overview on the history of this concept, this article will show how Papp drew on recent research by Otto Lehmann on liquid crystals to convey the idea that life may be based on other elements than carbon. The author concludes by arguing that popular science did not only make specialist knowledge accessible to a general public but also served to probe the limits of knowledge and point toward the situatedness of established categories and definitions. Key Words: History of astrobiology-Non-carbon-based life-Liquid crystals-Papp, Desiderius-Lehmann, Otto. Astrobiology 12, 951-957.

We analyzed tree rings in wood samples collected from some of the few surviving trees found close to the epicenter (within 4-5 km) of the Tunguska event that occurred on the last day of June 1908. Tree-ring growth shows a depression starting in the year after the event and continuing during a 4-5-year period. The most remarkable traces of the event were found in the rings' anatomical structure: (1) formation of "light" rings and a reduction of maximum density in 1908; (2) non-thickened tracheids (the cells that make up most of the wood volume) in the transition and latewood zones (the middle and last-formed parts of the ring, respectively); and (3) deformed tracheids, which are located on the 1908 annual ring outer boundary. In the majority of samples, normal earlywood and latewood tracheids were formed in all annual rings after 1908. The observed anomalies in wood anatomy suggest two main impacts of the Tunguska event on surviving trees--(1) defoliation and (2) direct mechanical stress on active xylem tissue. The mechanical stress needed to fell trees is less than the stress needed to cause the deformation of differentiating tracheids observed in trees close to the epicenter. In order to resolve this apparent contradiction, work is suggested on possible topographic modification of the overpressure experienced by these trees, as is an experimental test of the effects of such stresses on precisely analogous growing trees.

The Atacama along the Pacific Coast of Chile and Peru is one of the driest and possibly oldest deserts in the world. It represents an extreme habitat for life on Earth and is an analog for life in dry conditions on Mars. We report on four years (September 1994-October 1998) of climate and moisture data from the extreme arid region of the Atacama. Our data are focused on understanding moisture sources and their role in creating suitable environments for photosynthetic microorganisms in the desert surface. The average air temperature was 16.5 degrees C and 16.6 degrees C in 1995 and 1996, respectively. The maximum air temperature recorded was 37.9 degrees C, and the minimum was -5.7 degrees C. Annual average sunlight was 336 and 335 W m(-2) in 1995 and 1996, respectively. Winds averaged a few meters per second, with strong föhn winds coming from the west exceeding 12 m s(-1). During our 4 years of observation there was only one significant rain event of 2.3 mm, which occurred near midnight local time. We suggest that this event was a rainout of a heavy fog. It is of interest that the strong El Niño of 1997-1998 brought heavy rainfall to the deserts of Peru, but did not bring significant rain to the central Atacama in Chile. Dew occurred at our station frequently following high nighttime relative humidity, but is not a significant source of moisture in the soil or under stones. Groundwater also does not contribute to surface moisture. Only the one rain event of 2.3 mm resulted in liquid water in the soil and beneath stones for a total of only 65-85 h over 4 years. The paucity of liquid water under stones is consistent with the apparent absence of hypolithic (under-stone) cyanobacteria, the only known primary producers in such extreme deserts.

Abstract The future exploration of Mars will require access to the subsurface, along with acquisition of samples for scientific analysis and ground-truthing of water ice and mineral reserves for in situ resource utilization. The Icebreaker drill is an integral part of the Icebreaker mission concept to search for life in ice-rich regions on Mars. Since the mission targets Mars Special Regions as defined by the Committee on Space Research (COSPAR), the drill has to meet the appropriate cleanliness standards as requested by NASA's Planetary Protection Office. In addition, the Icebreaker mission carries life-detection instruments; and in turn, the drill and sample delivery system have to meet stringent contamination requirements to prevent false positives. This paper reports on the development and testing of the Icebreaker drill, a 1 m class rotary-percussive drill and triple redundant sample delivery system. The drill acquires subsurface samples in short, approximately 10 cm bites, which makes the sampling system robust and prevents thawing and phase changes in the target materials. Autonomous drilling, sample acquisition, and sample transfer have been successfully demonstrated in Mars analog environments in the Arctic and the Antarctic Dry Valleys, as well as in a Mars environmental chamber. In all environments, the drill has been shown to perform at the "1-1-100-100" level; that is, it drilled to 1 m depth in approximately 1 hour with less than 100 N weight on bit and approximately 100 W of power. The drilled substrate varied and included pure ice, ice-rich regolith with and without rocks and with and without 2% perchlorate, and whole rocks. The drill is currently at a Technology Readiness Level (TRL) of 5. The next-generation Icebreaker drill weighs 10 kg, which is representative of the flightlike model at TRL 5/6. Key Words: Drilling-Sampling-Mars-Mars drilling-Subsurface exploration-Ice-Search for life. Astrobiology 13, xxx-xxx.

Pioneering studies of Precambrian impact fallout units and associated tsunami deposits in the Hamersley Basin, Pilbara Craton, Western Australia, by B.M. Simonson and S.W. Hassler, document a range of tsunami deposits associated with impact fallout units whose impact connection is identified by associated microtektites and microkrystites (condensation spherules). The impact connection of these particles is demonstrated by iridium anomalies, unique platinum group elements patterns, and Ni-rich mineral phases. Densely packed tsunami-transported fragments and boulders overlie microkrystite units of the >2629 +/- 5 Ma top Jeerinah Impact Layer (JIL). Tsunami events closely follow spherule settling associated with the 2561 +/- 8 Ma Spherule Marker Bed SMB-1 and SMB-2 impact events, Bee Gorge Member, Wittenoom Formation. The two impact cycles are separated by a stratigraphically consistent silicified black siltstone, representing a "Quiet Interval." The SMB turbidites display turbulence eddies, climbing ripples, conglomerate pockets, slumps, and waterlogged sediment deformation features. Consequences of tsunami in the probably contemporaneous Carawine Dolomite (Pb-Pb carbonate ages of approximately 2.56-2.54 Ga), eastern Hamersley Basin, include sub-autochthonous below-wave base excavation and megabrecciation of sea floor substrata, resulting in a unique 10-30-m-thick spherule-bearing megabreccia marker mapped over a nearly 100-km north-south strike distance in the east Hamersley Basin. The field relations suggest a pretsunami settling of the bulk of the spherules. Tsunami wave effects include: (1). dispersal of the spherule-rich soft upper sea floor sediments as a subaqueous mud cloud and (2). excavation of consolidated substrata below the soft sediment zone. Excavation and megabrecciation included injection of liquefied spherule-bearing microbreccia into dilated fractures in the disrupted underlying carbonates. Near-perfect preservation of the spherules within the basal microbreccia veins suggests tsunami-induced hydraulic pressures locally exceeded lithostatic pressure. Late-stage settling of spherule-bearing mud clouds in the wake of the tsunami is represented by an abundance of spherules in the uppermost microbreccia zones of the megabreccia pile. From the deep below-wave base facies of the Carawine Dolomite, tsunami wave amplitudes may have exceeded 200 m depth. The approximately 2.47-2.50 Ga DGS4 (S4 Macroband, Dales Gorge Member, Brockman Iron Formation) fallout units include exotic chert and carbonate boulders transported by tsunami following settling of a 10-20-cm-thick microkrystite and microtektite-rich unit. Seismic perturbations preceding deposition of the JIL and SMB fallout units are marked by rip-up clasts. The geochemistry of microkrystites and microtektites suggests impact fallout originated from impacts in simatic/oceanic crustal regions, although tsunami waves may have originated from seismically reactivated faults and plate margins located at distance from the impact craters.

Apparent cyanobacterial emergence at about 2.8 Gya coincides with the negative excursion in the organic carbon isotope record, which is the first strong evidence for the presence of atmospheric methane. The existence of weathering feedbacks in the carbonate-silicate cycle suggests that atmospheric and oceanic CO2 concentrations would have been high prior to the presence of a methane greenhouse (and thus the ocean would have had high bicarbonate concentrations). With the onset of a methane greenhouse, carbon dioxide concentrations would decrease. Bicarbonate has been proposed as the preferred reductant that preceded water for oxygenic photosynthesis in a bacterial photosynthetic precursor to cyanobacteria; with the drop of carbon dioxide level, Archean cyanobacteria emerged using water as a reductant instead of bicarbonate (Dismukes et al., 2001). Our thermodynamic calculations, with regard to this scenario, give at least a tenfold drop in aqueous CO2 levels with the onset of a methane-dominated greenhouse, assuming surface temperatures of about 60 degrees C and a drop in the level of atmospheric carbon dioxide from about 1 to 0.1 bars. The buildup of atmospheric methane could have been triggered by the boost in oceanic organic productivity that arose from the emergence of pre-cyanobacterial oxygenic phototrophy at about 2.8-3.0 Gya; high temperatures may have precluded an earlier emergence. A greenhouse transition timescale on the order of 50-100 million years is consistent with results from modeling the carbonate-silicate cycle. This is an alternative hypothesis to proposals of a tectonic driver for this apparent greenhouse transition.

This experiment was aimed at establishing, for the first time, the survival capability of lichens exposed to space conditions. In particular, the damaging effect of various wavelengths of extraterrestrial solar UV radiation was studied. The lichens used were the bipolar species Rhizocarpon geographicum and Xanthoria elegans, which were collected above 2000 m in the mountains of central Spain and as endolithic communities inhabiting granites in the Antarctic Dry Valleys. Lichens were exposed to space in the BIOPAN-5 facility of the European Space Agency; BIOPAN-5 is located on the outer shell of the Earth-orbiting FOTON-M2 Russian satellite. The lichen samples were launched from Baikonur by a Soyuz rocket on May 31, 2005, and were returned to Earth after 16 days in space, at which time they were tested for survival. Chlorophyll fluorescence was used for the measurement of photosynthetic parameters. Scanning electron microscopy in back-scattered mode, low temperature scanning electron microscopy, and transmission electron microscopy were used to study the organization and composition of both symbionts. Confocal laser scanning microscopy, in combination with the use of specific fluorescent probes, allowed for the assessment of the physiological state of the cells. All exposed lichens, regardless of the optical filters used, showed nearly the same photosynthetic activity after the flight as measured before the flight. Likewise, the multimicroscopy approach revealed no detectable ultrastructural changes in most of the algal and fungal cells of the lichen thalli, though a greater proportion of cells in the flight samples had compromised membranes, as revealed by the LIVE/DEAD BacLight Bacterial Viability Kit. These findings indicate that most lichenized fungal and algal cells can survive in space after full exposure to massive UV and cosmic radiation, conditions proven to be lethal to bacteria and other microorganisms. The lichen upper cortex seems to provide adequate protection against solar radiation. Moreover, after extreme dehydration induced by high vacuum, the lichens proved to be able to recover, in full, their metabolic activity within 24 hours.

Sampling of subsurface rock may be required to detect evidence of past biological activity on Mars. The Mars Astrobiology Research and Technology Experiment (MARTE) utilized the Río Tinto region, Spain, as a Mars analog site to test dry drilling technologies specific to Mars that retrieve subsurface rock for biological analysis. This work examines the usefulness of visible-near infrared (VNIR) (450-1000 nm) point spectrometry to characterize ferric iron minerals in core material retrieved during a simulated Mars drilling mission. VNIR spectrometry can indicate the presence of aqueously precipitated ferric iron minerals and, thus, determine whether biological analysis of retrieved rock is warranted. Core spectra obtained during the mission with T1 (893-897 nm) and T2 (644-652 nm) features indicate goethite-dominated samples, while relatively lower wavelength T1 (832-880 nm) features indicate hematite. Hematite/goethite molar ratios varied from 0 to 1.4, and within the 880-898 nm range, T1 features were used to estimate hematite/goethite molar ratios. Post-mission X-ray analysis detected phyllosilicates, which indicates that examining beyond the VNIR (e.g., shortwave infrared, 1000-2500 nm) will enhance the detection of other minerals formed by aqueous processes. Despite the limited spectral range of VNIR point spectrometry utilized in the MARTE Mars drilling simulation project, ferric iron minerals could be identified in retrieved core material, and their distribution served to direct core subsampling for biological analysis.

The Mars Astrobiology Research and Technology Experiment (MARTE) simulated a robotic drilling mission to search for subsurface life on Mars. The drill site was on Peña de Hierro near the headwaters of the Río Tinto river (southwest Spain), on a deposit that includes massive sulfides and their gossanized remains that resemble some iron and sulfur minerals found on Mars. The mission used a fluidless, 10-axis, autonomous coring drill mounted on a simulated lander. Cores were faced; then instruments collected color wide-angle context images, color microscopic images, visible-near infrared point spectra, and (lower resolution) visible-near infrared hyperspectral images. Cores were then stored for further processing or ejected. A borehole inspection system collected panoramic imaging and Raman spectra of borehole walls. Life detection was performed on full cores with an adenosine triphosphate luciferin-luciferase bioluminescence assay and on crushed core sections with SOLID2, an antibody array-based instrument. Two remotely located science teams analyzed the remote sensing data and chose subsample locations. In 30 days of operation, the drill penetrated to 6 m and collected 21 cores. Biosignatures were detected in 12 of 15 samples analyzed by SOLID2. Science teams correctly interpreted the nature of the deposits drilled as compared to the ground truth. This experiment shows that drilling to search for subsurface life on Mars is technically feasible and scientifically rewarding.

The Astrobiology Primer has been created as a reference tool for those who are interested in the interdisciplinary field of astrobiology. The field incorporates many diverse research endeavors, but it is our hope that this slim volume will present the reader with all he or she needs to know to become involved and to understand, at least at a fundamental level, the state of the art. Each section includes a brief overview of a topic and a short list of readable and important literature for those interested in deeper knowledge. Because of the great diversity of material, each section was written by a different author with a different expertise. Contributors, authors, and editors are listed at the beginning, along with a list of those chapters and sections for which they were responsible. We are deeply indebted to the NASA Astrobiology Institute (NAI), in particular to Estelle Dodson, David Morrison, Ed Goolish, Krisstina Wilmoth, and Rose Grymes for their continued enthusiasm and support. The Primer came about in large part because of NAI support for graduate student research, collaboration, and inclusion as well as direct funding. We have entitled the Primer version 1 in hope that it will be only the first in a series, whose future volumes will be produced every 3-5 years. This way we can insure that the Primer keeps up with the current state of research. We hope that it will be a great resource for anyone trying to stay abreast of an ever-changing field.

The Tardigrade Resistance to Space Effects (TARSE) project, part of the mission LIFE on FOTON-M3, analyzed the effects of the space environment on desiccated and active tardigrades. Four experiments were conducted in which the eutardigrade Macrobiotus richtersi was used as a model species. Desiccated (in leaf litter or on paper) and hydrated tardigrades (fed or starved) were flown on FOTON-M3 for 12 days in September 2007, which, for the first time, allowed for a comparison of the effects of the space environment on desiccated and on active animals. In this paper, we report the experimental design of the TARSE project and data on tardigrade survival. In addition, data on survival, genomic DNA integrity, Hsp70 and Hsp90 expressions, antioxidant enzyme contents and activities, and life history traits were compared between hydrated starved tardigrades flown in space and those maintained on Earth as a control. Microgravity and radiation had no effect on survival or DNA integrity of active tardigrades. Hsp expressions between the animals in space and the control animals on Earth were similar. Spaceflight induced an increase of glutathione content and its related enzymatic activities. Catalase and superoxide dismutase decreased with spaceflight, and thiobarbituric acid reactive substances did not change. During the flight mission, tardigrades molted, and females laid eggs. Several eggs hatched, and the newborns exhibited normal morphology and behavior.

We present a summary of the physical principles and design of the Dynamic Albedo of Neutrons (DAN) instrument onboard NASA's 2009 Mars Science Laboratory (MSL) mission. The DAN instrument will use the method of neutron-neutron activation analysis in a space application to study the abundance and depth distribution of water in the martian subsurface along the path of the MSL rover.

The Mars Program Plan includes an integrated and coordinated set of future candidate missions and investigations that meet fundamental science objectives of NASA and the Mars Exploration Program (MEP). At the time this paper was written, these possible future missions are planned in a manner consistent with a projected budget profile for the Mars Program in the next decade (2007-2016). As with all future missions, the funding profile depends on a number of factors that include the exact cost of each mission as well as potential changes to the overall NASA budget. In the current version of the Mars Program Plan, the Astrobiology Field Laboratory (AFL) exists as a candidate project to determine whether there were (or are) habitable zones and life, and how the development of these zones may be related to the overall evolution of the planet. The AFL concept is a surface exploration mission equipped with a major in situ laboratory capable of making significant advancements toward the Mars Program's life-related scientific goals and the overarching Vision for Space Exploration. We have developed several concepts for the AFL that fit within known budget and engineering constraints projected for the 2016 and 2018 Mars mission launch opportunities. The AFL mission architecture proposed here assumes maximum heritage from the 2009 Mars Science Laboratory (MSL). Candidate payload elements for this concept were identified from a set of recommendations put forth by the Astrobiology Field Laboratory Science Steering Group (AFL SSG) in 2004, for the express purpose of identifying overall rover mass and power requirements for such a mission. The conceptual payload includes a Precision Sample Handling and Processing System that would replace and augment the functionality and capabilities provided by the Sample Acquisition Sample Processing and Handling system that is currently part of the 2009 MSL platform.

In this study, we utilized transmission electron microscopy to examine the contents of fluid inclusions in halite (NaCl) and solid halite crystals collected 650 m below the surface from the Late Permian Salado Formation in southeastern New Mexico (USA). The halite has been isolated from contaminating groundwater since deposition approximately 250 Ma ago. We show that abundant cellulose microfibers are present in the halite and appear remarkably intact. The cellulose is in the form of 5 nm microfibers as well as composite ropes and mats, and was identified by resistance to 0.5 N NaOH treatment and susceptibility to cellulase enzyme treatment. These cellulose microfibers represent the oldest native biological macromolecules to have been directly isolated, examined biochemically, and visualized (without growth or replication) to date. This discovery points to cellulose as an ideal macromolecular target in the search for life on other planets in our Solar System.

Modern stromatolites represent a significant resource for studying microbial ecology and evolution. A preliminary investigation was undertaken employing specific genetic probes to characterize the cyanobacteria responsible for stromatolite construction in a range of environments, including microbial mats found in Australia not previously examined with molecular methods. Isolates of cyanobacteria were collected from stromatolites in thermal springs, hypersaline lakes, and oceanic fringes on two continents. A polymerase chain reaction specific for DNA of cyanobacterial 16S rRNA was developed, the resulting products of the DNA amplification reaction were sequenced, and the data were used to infer relatedness between the isolates studied and other members of the cyanobacterial radiation. Complete sequence was generated for the region from position 27 to 408 for 13 strains of cyanobacteria associated with stromatolites. All stromatolite-derived sequences were most closely related to cyanobacteria, as indicated by local sequence alignment. It was possible to correlate genetic identity with morphological nomenclatures and to expand the phylogeny of benthic cyanobacteria. These inferences were also expanded to temporal variation in the dominant resident cyanobacterial species based on sampling of surface and core sinter laminations. Under the methods employed, only one cyanobacterial strain was detected in each sample, suggesting the possible dominance of a specific clonal population of cyanobacteria at any one time in the biota of the samples tested. The data indicate that internal core samples of a stromatolite at least 10 years old can be successfully analyzed by DNA-based methods to identify preserved cyanobacteria.

Herein, we report on the purification, characterization, and sequencing of catalase from Acinetobacter gyllenbergii 2P01AA, an extremely oxidation-resistant bacterium that was isolated from the Mars Phoenix spacecraft assembly facility. The Acinetobacter are dominant members of the microbial communities that inhabit spacecraft assembly facilities and consequently may serve as forward contaminants that could impact the integrity of future life-detection missions. Catalase was purified by using a 3-step chromatographic procedure, where mass spectrometry provided respective subunit and intact masses of 57.8 and 234.6 kDa, which were consistent with a small-subunit tetrameric catalase. Kinetics revealed an extreme pH stability with no loss in activity between pH 5 and 11.5 and provided respective kcat/Km and kcat values of ∼10(7) s(-1) M(-1) and 10(6) s(-1), which are among the highest reported for bacterial catalases. The amino acid sequence was deduced by in-depth peptide mapping, and structural homology suggested that the catalases from differing strains of A. gyllenbergii differ only at residues near the subunit interfaces, which may impact catalytic stability. Together, the kinetic, alkali-tolerant, and halotolerant properties of the catalase from A. gyllenbergii 2P01AA are significant, as they are consistent with molecular adaptations toward the alkaline, low-humidity, and potentially oxidizing conditions of spacecraft assembly facilities. Therefore, these results support the hypothesis that the selective pressures of the assembly facilities impact the microbial communities at the molecular level, which may have broad implications for future life-detection missions. Key Words: Catalase-Kinetics-Sequence-Stability-Acinetobacter-Spacecraft-Planetary protection. Astrobiology 15, xxx-xxx.

Four layers, S1-S4, containing sand-sized spherical particles formed as a result of large meteorite impacts, occur in 3.47-3.24 Ga rocks of the Barberton Greenstone Belt, South Africa. Ir levels in S3 and S4 locally equal or exceed chondritic values but in other sections are at or only slightly above background. Most spherules are inferred to have formed by condensation of impact-produced rock vapor clouds, although some may represent ballistically ejected liquid droplets. Extreme Ir abundances and heterogeneity may reflect element fractionation during spherule formation, hydraulic fractionation during deposition, and/or diagenetic and metasomatic processes. Deposition of S1, S2, and S3 was widely influenced by waves and/or currents interpreted to represent impact-generated tsunamis, and S1 and S2 show multiple graded layers indicating the passage of two or more wave trains. These tsunamis may have promoted mixing within a globally stratified ocean, enriching surface waters in nutrients for biological communities. S2 and S3 mark the transition from the 300-million-year-long Onverwacht stage of predominantly basaltic and komatiitic volcanism to the late orogenic stage of greenstone belt evolution, suggesting that regional and possibly global tectonic reorganization resulted from these large impacts. These beds provide the oldest known direct record of terrestrial impacts and an opportunity to explore their influence on early life, crust, ocean, and atmosphere. The apparent presence of impact clusters at 3.26-3.24 Ga and approximately 2.65-2.5 Ga suggests either spikes in impact rates during the Archean or that the entire Archean was characterized by terrestrial impact rates above those currently estimated from the lunar cratering record.

Three morphotypes of microbial mats are preserved in rocks deposited in shallow-water facies of the 3.42 Ga Buck Reef chert (BRC). Morphotype alpha consists of fine anastomosing and bifurcating carbonaceous laminations, which loosely drape underlying detrital grains or form silica-filled lenses. Morphotype beta consists of meshes of fine carbonaceous strands intergrown with detrital grains and dark laminations, which loosely drape coarse detrital grains. Morphotype gamma consists of fine, even carbonaceous laminations that tightly drape underlying detrital grains. Preservation of nearly uncompacted mat morphologies and detrital grains deposited during mat growth within a well-characterized sedimentary unit makes quantitative correlation between morphology and paleoenvironment possible. All mats are preserved in the shallowest-water interval of those rocks deposited below normal wave base and above storm wave base. This interval is bounded below by a transgressive lag formed during regional flooding and above by a small condensed section that marks a local relative sea-level maximum. Restriction of all mat morphotypes to the shallowest interval of the storm-active layer in the BRC ocean reinforces previous interpretations that these mats were constructed primarily by photosynthetic organisms. Morphotypes alpha and beta dominate the lower half of this interval and grew during deposition of relatively coarse detrital carbonaceous grains, while morphotype gamma dominates the upper half and grew during deposition of fine detrital carbonaceous grains. The observed mat distribution suggests that either light intensity or, more likely, small variations in ambient current energy acted as a first-order control on mat morphotype distribution. These results demonstrate significant environmental control on biological morphogenetic processes independent of influences from siliciclastic sedimentation.

Abstract Microbially induced sedimentary structures (MISS) result from the response of microbial mats to physical sediment dynamics. MISS are cosmopolitan and found in many modern environments, including shelves, tidal flats, lagoons, riverine shores, lakes, interdune areas, and sabkhas. The structures record highly diverse communities of microbial mats and have been reported from numerous intervals in the geological record up to 3.2 billion years (Ga) old. This contribution describes a suite of MISS from some of the oldest well-preserved sedimentary rocks in the geological record, the early Archean (ca. 3.48 Ga) Dresser Formation, Western Australia. Outcrop mapping at the meter to millimeter scale defined five sub-environments characteristic of an ancient coastal sabkha. These sub-environments contain associations of distinct macroscopic and microscopic MISS. Macroscopic MISS include polygonal oscillation cracks and gas domes, erosional remnants and pockets, and mat chips. Microscopic MISS comprise tufts, sinoidal structures, and laminae fabrics; the microscopic laminae are composed of primary carbonaceous matter, pyrite, and hematite, plus trapped and bound grains. Identical suites of MISS occur in equivalent environmental settings through the entire subsequent history of Earth including the present time. This work extends the geological record of MISS by almost 300 million years. Complex mat-forming microbial communities likely existed almost 3.5 billion years ago. Key Words: Archean-Biofilms-Microbial mats-Early Earth-Evolution. Astrobiology 13, xxx-xxx.

Abstract Sandstone beds of the <3.7 Ga Gillespie Lake Member on Mars have been interpreted as evidence of an ancient playa lake environment. On Earth, such environments have been sites of colonization by microbial mats from the early Archean to the present time. Terrestrial microbial mats in playa lake environments form microbialites known as microbially induced sedimentary structures (MISS). On Mars, three lithofacies of the Gillespie Lake Member sandstone display centimeter- to meter-scale structures similar in macroscopic morphology to terrestrial MISS that include "erosional remnants and pockets," "mat chips," "roll-ups," "desiccation cracks," and "gas domes." The microbially induced sedimentary-like structures identified in Curiosity rover mission images do not have a random distribution. Rather, they were found to be arranged in spatial associations and temporal successions that indicate they changed over time. On Earth, if such MISS occurred with this type of spatial association and temporal succession, they would be interpreted as having recorded the growth of a microbially dominated ecosystem that thrived in pools that later dried completely: erosional pockets, mat chips, and roll-ups resulted from water eroding an ancient microbial mat-covered sedimentary surface; during the course of subsequent water recess, channels would have cut deep into the microbial mats, leaving erosional remnants behind; desiccation cracks and gas domes would have occurred during a final period of subaerial exposure of the microbial mats. In this paper, the similarities of the macroscopic morphologies, spatial associations, and temporal succession of sedimentary structures on Mars to MISS preserved on Earth has led to the following hypothesis: The sedimentary structures in the <3.7 Ga Gillespie Lake Member on Mars are ancient MISS produced by interactions between microbial mats and their environment. Proposed here is a strategy for detecting, identifying, confirming, and differentiating possible MISS during current and future Mars missions. Key Words: Astrobiology-Life on Mars-Microbial mats-MISS-Biosignature-Curiosity rover. Astrobiology 15, xxx-xxx.

Abstract Several lines of evidence indicate the existence of salty liquid water below the icy surface of the satellite Europa. Depending on the chemical composition of the original interior brines, minerals that precipitate will be varied as will be the resulting physicochemical parameters of the evolving solutions such as pH and salinity. These parameters are determinants apropos to the study of the possible habitability of the satellite. In this work, experiments of fractional precipitation by cooling of several brines with different chemical composition (acid, alkaline, and neutral) were performed at 1 and 300 bar. The gradual decrease in temperature leads to mineral precipitation and changes in salinity and pH values. During the experiment, Raman spectroscopy was used to analyze quantitatively the variation of the salt concentration in the aqueous solutions. The obtained laboratory data indicate the manner in which cryomagma differentiation might occur on Europa. These endogenous processes of differentiation require planetary energy, which seems to have been plentiful during Europa's geological history. Ultimately, the dissipation of part of that energy is translated to a higher complexity of the cryopetrology in Europa's crust. From the results, we conclude that fractional differentiation processes of briny cryomagmas produce several types of igneous salty mineral suites on icy moons. Key Words: Europa-Salts-Chemical evolution-Raman spectroscopy-Volcanism. Astrobiology 13, 693-702.