With a grain of salt: what halite has to offer to discussions on the origin of life

This experimental study investigated how the dynamics of the crystallization of the evaporite mineral halite could affect the accumulation and preservation of organic macromolecules present in the crystallizing solution. Halite was grown under controlled conditions in the presence of polymer nanoparticles that acted as an analog to protocellular material. Optical microscopy, atomic force microscopy, and laser scanning confocal fluorescence microscopy were used to trace the localization of the nanoparticles during and after growth of halite crystals. The present study revealed that the organic nanoparticles were not regularly incorporated within the halite, but were very concentrated on its surfaces. Their distribution was controlled dominantly by the morphologic surface features of the mineral rather than by specific molecular interactions with an atomic plane of the mineral. This means that the distribution of organic molecules was controlled by surfaces like those of halite's evaporitic growth forms. The experiments with halite also demonstrated that a mineral need not continuously incorporate organic molecules during its crystallization to preserve those molecules: After rejection by (non-incorporation into) the crystallizing halite, the organic nanoparticles increased in concentration in the evaporating brine. They ultimately either adsorbed in rectilinear patterns onto the hopper-enhanced surfaces and along discontinuities within the crystals, or they were encapsulated within fluid inclusions. Of additional importance in origin-of-life considerations is the fact that halite in the natural environment rapidly can change its role from that of a protective repository (in the absence of water) to that of a source of organic particles (as soon as water is present) when the mineral dissolves.     

"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.     

A microbial oasis in the hypersaline Atacama subsurface discovered by a life detector chip: implications for the search for life on Mars

The Atacama Desert has long been considered a good Mars analogue for testing instrumentation for planetary exploration, but very few data (if any) have been reported about the geomicrobiology of its salt-rich subsurface. We performed a Mars analogue drilling campaign next to the Salar Grande (Atacama, Chile) in July 2009, and several cores and powder samples from up to 5?m deep were analyzed in situ with LDChip300 (a Life Detector Chip containing 300 antibodies). Here, we show the discovery of a hypersaline subsurface microbial habitat associated with halite-, nitrate-, and perchlorate-containing salts at 2?m deep. LDChip300 detected bacteria, archaea, and other biological material (DNA, exopolysaccharides, some peptides) from the analysis of less than 0.5?g of ground core sample. The results were supported by oligonucleotide microarray hybridization in the field and finally confirmed by molecular phylogenetic analysis and direct visualization of microbial cells bound to halite crystals in the laboratory. Geochemical analyses revealed a habitat with abundant hygroscopic salts like halite (up to 260?g kg(-1)) and perchlorate (41.13?μg g(-1) maximum), which allow deliquescence events at low relative humidity. Thin liquid water films would permit microbes to proliferate by using detected organic acids like acetate (19.14?μg g(-1)) or formate (76.06?μg g(-1)) as electron donors, and sulfate (15875?μg g(-1)), nitrate (13490?μg g(-1)), or perchlorate as acceptors. Our results correlate with the discovery of similar hygroscopic salts and possible deliquescence processes on Mars, and open new search strategies for subsurface martian biota. The performance demonstrated by our LDChip300 validates this technology for planetary exploration, particularly for the search for life on Mars.     

Substrate limitation for methanogenesis in hypersaline environments

Motivated by the increasingly abundant evidence for hypersaline environments on Mars and reports of methane in its atmosphere, we examined methanogenesis in hypersaline ponds in Baja California Sur, Mexico, and in northern California, USA. Methane-rich bubbles trapped within or below gypsum/halite crusts have δ13C values near -40‰. Methane with these relatively high isotopic values would typically be considered thermogenic; however, incubations of crust samples resulted in the biological production of methane with similar isotopic composition. A series of measurements aimed at understanding the isotopic composition of methane in hypersaline systems was therefore undertaken. Methane production rates, as well as the concentrations and isotopic composition of the particulate organic carbon (POC), were measured. Methane production was highest from microbial communities living within gypsum crusts, whereas POC content at gypsum/halite sites was low, generally less than 1% of the total mass. The isotopic composition of the POC ranged from -26‰ to -10‰. To determine the substrates used by the methanogens, 13C-labeled methylamines, methanol, acetate, and bicarbonate were added to individual incubation vials, and the methane produced was monitored for 13C content. The main substrates used by the methanogens were the noncompetitive substrates, the methylamines, and methanol. When unlabeled trimethylamine (TMA) was added to incubating gypsum/halite crusts in increasing concentrations, the isotopic composition of the methane produced became progressively lower; the lowest methane δ13C values occurred when the most TMA was added (1000 μM final concentration). This decrease in the isotopic composition of the methane produced with increasing TMA concentrations, along with the high in situ methane δ13C values, suggests that the methanogens within the crusts are operating at low substrate concentrations. It appears that substrate limitation is decreasing isotopic fractionation during methanogenesis, which results in these abnormally high biogenic methane δ13C values.     

Entrapment of bacteria in fluid inclusions in laboratory-grown halite

Cells of the bacterium Pseudomonas aeruginosa, which were genetically modified to produce green fluorescent protein, were entrapped in fluid inclusions in laboratory-grown halite. The bacteria were used to inoculate NaCl-saturated aqueous solutions, which were allowed to evaporate and precipitate halite. The number, size, and distribution of fluid inclusions were highly variable, but did not appear to be affected by the presence of the bacteria. Many of the inclusions in crystals from inoculated solutions contained cells in populations ranging from two to 20. Microbial attachment to crystal surfaces was neither evident nor necessary for entrapment. Cells occurred exclusively within fluid inclusions and were not present in the crystal matrix. In both the inclusions and the hypersaline solution, the cells fluoresced and twitched, which indicates that the bacteria might have remained viable after entrapment. The fluorescence continued up to 13 months after entrapment, which indicates that little degradation of the bacteria occurred over that time interval. The entrapment, fluorescence, and preservation of cells were independent of the volume of hypersaline solution used or whether the solutions were completely evaporated prior to crystal extraction. The results of this study have a wide range of implications for the long-term survival of microorganisms in fluid inclusions and their detection through petrography. The results also demonstrate the preservation potential for microbes in hypersaline fluid inclusions, which could allow cells to survive harsh conditions of space, the deep geologic past, or burial in sedimentary basins.     

Responses of haloarchaea to simulated microgravity

Various effects of microgravity on prokaryotes have been recognized in recent years, with the focus on studies of pathogenic bacteria. No archaea have been investigated yet with respect to their responses to microgravity. For exposure experiments on spacecrafts or on the International Space Station, halophilic archaea (haloarchaea) are usually embedded in halite, where they accumulate in fluid inclusions. In a liquid environment, these cells will experience microgravity in space, which might influence their viability and survival. Two haloarchaeal strains, Haloferax mediterranei and Halococcus dombrowskii, were grown in simulated microgravity (SMG) with the rotary cell culture system (RCCS, Synthecon). Initially, salt precipitation and detachment of the porous aeration membranes in the RCCS were observed, but they were avoided in the remainder of the experiment by using disposable instead of reusable vessels. Several effects were detected, which were ascribed to growth in SMG: Hfx. mediterranei's resistance to the antibiotics bacitracin, erythromycin, and rifampicin increased markedly; differences in pigmentation and whole cell protein composition (proteome) of both strains were noted; cell aggregation of Hcc. dombrowskii was notably reduced. The results suggest profound effects of SMG on haloarchaeal physiology and cellular processes, some of which were easily observable and measurable. This is the first report of archaeal responses to SMG. The molecular mechanisms of the effects induced by SMG on prokaryotes are largely unknown; haloarchaea could be used as nonpathogenic model systems for their elucidation and in addition could provide information about survival during lithopanspermia (interplanetary transport of microbes inside meteorites).     

Carotenoid analysis of halophilic archaea by resonance Raman spectroscopy

Recently, halite and sulfate evaporate rocks have been discovered on Mars by the NASA rovers, Spirit and Opportunity. It is reasonable to propose that halophilic microorganisms could have potentially flourished in these settings. If so, biomolecules found in microorganisms adapted to high salinity and basic pH environments on Earth may be reliable biomarkers for detecting life on Mars. Therefore, we investigated the potential of Resonance Raman (RR) spectroscopy to detect biomarkers derived from microorganisms adapted to hypersaline environments. RR spectra were acquired using 488.0 and 514.5 nm excitation from a variety of halophilic archaea, including Halobacterium salinarum NRC-1, Halococcus morrhuae, and Natrinema pallidum. It was clearly demonstrated that RR spectra enhance the chromophore carotenoid molecules in the cell membrane with respect to the various protein and lipid cellular components. RR spectra acquired from all halophilic archaea investigated contained major features at approximately 1000, 1152, and 1505 cm(-1). The bands at 1505 cm(-1) and 1152 cm(-1) are due to in-phase C=C (nu(1) ) and C-C stretching ( nu(2) ) vibrations of the polyene chain in carotenoids. Additionally, in-plane rocking modes of CH(3) groups attached to the polyene chain coupled with C-C bonds occur in the 1000 cm(-1) region. We also investigated the RR spectral differences between bacterioruberin and bacteriorhodopsin as another potential biomarker for hypersaline environments. By comparison, the RR spectrum acquired from bacteriorhodopsin is much more complex and contains modes that can be divided into four groups: the C=C stretches (1600-1500 cm(-1)), the CCH in-plane rocks (1400-1250 cm(-1)), the C-C stretches (1250-1100 cm(-1)), and the hydrogen out-of-plane wags (1000-700 cm(-1)). RR spectroscopy was shown to be a useful tool for the analysis and remote in situ detection of carotenoids from halophilic archaea without the need for large sample sizes and complicated extractions, which are required by analytical techniques such as high performance liquid chromatography and mass spectrometry.     

Tírez lake as a terrestrial analog of Europa

Tírez Lake (La Mancha, central Spain) is proposed as a terrestrial analogue of Europa's ocean. The proposal is based on the comparison of the hydrogeochemistry of Tírez Lake with the geochemical features of the alteration mineralogy of meteoritic precursors and with Galileo's Near Infrared Mapping Spectrometer data on Europa's surface. To validate the astrobiological potential of Tírez Lake as an analog of Europa, different hydrogeochemical, mineral, and microbial analyses were performed. Experimental and theoretical modeling helped to understand the crystallization pathways that may occur in Europa's crust. Calculations about the oxidation state of the hypothetical Europan ocean were estimated to support the sulfate-rich neutral liquid model as the origin of Europa's observed hydrated minerals and to facilitate their comparison with Tírez's hydrogeochemistry. Hydrogeochemical and mineralogical analyses showed that Tírez waters corresponded to Mg-Na-SO(4)-Cl brines with epsomite, hexahydrite, and halite as end members. A preliminary microbial ecology characterization identified two different microbial domains: a photosynthetically sustained community represented by planktonic/benthonic forms and microbial mat communities, and a subsurficial anaerobic realm in which chemolithotrophy predominates. Fluorescence in situ hybridization has been used to characterize the prokaryotic diversity of the system. The subsurficial community seemed to be dominated by sulfate-reducing bacteria and methanogens. Frozen Tírez brines were analyzed by Fourier-transform infrared techniques providing spectra similar to those reported previously using pure components and to the Galileo spectral data. Calorimetric measurements of Tírez brines showed pathways and phase metastability for magnesium sulfate and sodium chloride crystallization that may aid in understanding the processes involved in the formation of Europa's icy crust. The use of fluorescence hybridization techniques for microbial detection and characterization in hypersaline environments makes this methodology strongly advisable for future Europa astrobiological missions.     

Endolithic cyanobacteria in halite rocks from the hyperarid core of the Atacama Desert

In the driest parts of the Atacama Desert there are no visible life forms on soil or rock surfaces. The soil in this region contains only minute traces of bacteria distributed in patches, and conditions are too dry for cyanobacteria that live under translucent stones. Here we show that halite evaporite rocks from the driest part of the Atacama Desert are colonized by cyanobacteria. This colonization takes place just a few millimeters beneath the rock surface, occupying spaces among salt crystals. Our work reveals that these communities are composed of extremely resistant Chroococcidiopsis morphospecies of cyanobacteria and associated heterotrophic bacteria. This newly discovered endolithic environment is an extremely dry and, at the same time, saline microbial habitat. Photosynthetic microorganisms within dry evaporite rocks could be an important and previously unrecognized target for the search for life within our Solar System.     

Bioconversion systems for food and water on long term space missions

An imperative for prolonged Space flight missions is the conservation of resources. Extensive resupply could pose technological and logistical challenges for those responsible for the management and successful completion of the mission. Therefore, the biological waste water reclamation system (BWWR) which requires little or no expendable supplies and the waste cellulose to edible mushroom conversion system (CMCS) which is conceived as a low energy crop waste recycling system are prototype instruments which have been conceived as solutions to the mission resupply problem. Out tests, conducted with relatively crude devices based on the original concepts, indicate that further research on the basic principles underlying the systems and refinement of the engineering designs will lead to hardware with the potential to satisfy the requirement for minimal re-supply while providing recycled water and edible mushrooms.     

Supercooled water brines within permafrost-an unknown ecological niche for microorganisms: a model for astrobiology

This study describes brine lenses (cryopegs) found in Siberian permafrost derived from ancient marine sediment layers of the Arctic Ocean. The cryopegs were formed and isolated from sediment ~100,000-120,000 years ago. They remain liquid at the in situ temperature of -10 degrees C as a result of their high salt content (170-300 g/L). [(14)C] Glucose is taken up by the cryopeg biomass at -15 degrees C, indicating microbial metabolism at low temperatures in this habitat. Furthermore, aerobic, anaerobic heterotrophs, sulfate reducers, acetogens, and methanogens were detected by most probable number analysis. Two psychrophilic microbes were isolated from the cryopegs, a Clostridium and a Psychrobacter. The closest relatives of each were previously isolated from Antarctica. The cryopeg econiche might serve as a model for extraterrestrial life, and hence is of particular interest to astrobiology.     

Mutagenesis in bacterial spores exposed to space and simulated martian conditions: data from the EXPOSE-E spaceflight experiment PROTECT

As part of the PROTECT experiment of the EXPOSE-E mission on board the International Space Station (ISS), the mutagenic efficiency of space was studied in spores of Bacillus subtilis 168. After 1.5 years' exposure to selected parameters of outer space or simulated martian conditions, the rates of induced mutations to rifampicin resistance (Rif(R)) and sporulation deficiency (Spo(-)) were quantified. In all flight samples, both mutations, Rif(R) and Spo(-), were induced and their rates increased by several orders of magnitude. Extraterrestrial solar UV radiation (>110?nm) as well as simulated martian UV radiation (>200?nm) led to the most pronounced increase (up to nearly 4 orders of magnitude); however, mutations were also induced in flight samples shielded from insolation, which were exposed to the same conditions except solar irradiation. Nucleotide sequencing located the Rif(R) mutations in the rpoB gene encoding the β-subunit of RNA polymerase. Mutations isolated from flight and parallel mission ground reference (MGR) samples were exclusively localized to Cluster I. The 21 Rif(R) mutations isolated from the flight experiment showed all a C to T transition and were all localized to one hotspot: H482Y. In mutants isolated from the MGR, the spectrum was wider with predicted amino acid changes at residues Q469K/L/R, H482D/P/R/Y, and S487L. The data show the unique mutagenic power of space and martian surface conditions as a consequence of DNA injuries induced by solar UV radiation and space vacuum or the low pressure of Mars.     

纤维素甘油醚硝酸酯在双基推进剂中的应用

纤维素甘油醚硝酸酯NGEC是一种改性硝化棉,以NGEC为粘合剂代替NC,采用传统的无溶剂法制备了NGEC基改性双基推进剂。研究了以NGEC为粘合剂的改性双基推进剂的力学、燃烧和弹道性能及燃烧波结构。结果表明,NGEC具有较好的热塑性,NGEC基改性双基推进剂加工性能优良,其低温-40℃延伸率可达相同配方NC基改性双基推进剂的1.5倍以上,其推进剂弹道性能稳定,燃烧性能和燃烧波结构与NC基推进剂相似。因此,NGEC作为含能粘合剂代替NC,可用于改善改性双基推进剂性能的研究。     

Spore UV and acceleration resistance of endolithic Bacillus pumilus and Bacillus subtilis isolates obtained from Sonoran desert basalt: implications for lithopanspermia

Bacterial spores have been used as model systems for studying the theory of interplanetary transport of life by natural processes such as asteroidal or cometary impacts (i.e., lithopanspermia). Because current spallation theory predicts that near-surface rocks are ideal candidates for planetary ejection and surface basalts are widely distributed throughout the rocky planets, we isolated spore-forming bacteria from the interior of near-subsurface basalt rocks collected in the Sonoran desert near Tucson, Arizona. Spores were found to inhabit basalt at very low concentrations (相似文献   

Microbial populations in Antarctic permafrost: biodiversity, state, age, and implication for astrobiology

Antarctic permafrost soils have not received as much geocryological and biological study as has been devoted to the ice sheet, though the permafrost is more stable and older and inhabited by more microbes. This makes these soils potentially more informative and a more significant microbial repository than ice sheets. Due to the stability of the subsurface physicochemical regime, Antarctic permafrost is not an extreme environment but a balanced natural one. Up to 10(4) viable cells/g, whose age presumably corresponds to the longevity of the permanently frozen state of the sediments, have been isolated from Antarctic permafrost. Along with the microbes, metabolic by-products are preserved. This presumed natural cryopreservation makes it possible to observe what may be the oldest microbial communities on Earth. Here, we describe the Antarctic permafrost habitat and biodiversity and provide a model for martian ecosystems.     

Aspartic acid racemization and age-depth relationships for organic carbon in Siberian permafrost

We have analyzed the degree of racemization of aspartic acid in permafrost samples from Northern Siberia, an area from which microorganisms of apparent ages up to a few million years have previously been isolated and cultured. We find that the extent of aspartic acid racemization in permafrost cores increases very slowly up to an age of approximately 25,000 years (around 5 m in depth). The apparent temperature of racemization over the age range of 0-25,000 years, determined using measured aspartic acid racemization rate constants, is -19 degrees C. This apparent racemization temperature is significantly lower than the measured environmental temperature (-11 to -13 degrees C) and suggests active recycling of D-aspartic acid in Siberian permafrost up to an age of around 25,000 years. This indicates that permafrost organisms are capable of repairing some molecular damage incurred while in a "dormant" state over geologic time.     

Assessment of space environmental factors by cytotoxicity bioassays

Cellular bioassays for detection of cyto- and genotoxicity are useful in the risk assessment of space environmental factors. Such bioassay systems have the potential complement the physical detector systems used in space, insofar as they yield intrinsically biologically weighted measures of cellular responses. The experiment Cellular Responses to Radiation in Space (CERASP) has been selected by NASA/ESA to be performed on the International Space Station. It will supply basic information on the cellular response to radiation applied in microgravity. One of the biological endpoints under investigation will be survival reflected by radiation-dependent reduction of constitutive expression of the enhanced variant of green fluorescent protein (EGFP), originally isolated from the bioluminescent jellyfish Aequorea victoria. In this ground based study, the usefulness of this approach in comparison to standard techniques (colony forming ability test, MTT test) is shown.     

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.     

Real time confocal laser scanning microscopy: potential applications in space medicine and cell biology

Photodynamic therapy (PDT), in which tissues may be rendered fatally light-sensitive represents a relatively novel treatment for cancer and other disorders such as cardiovascular disease. It offers significant application to disease control in an isolated environment such as space flight. In studying PDT in the laboratory, low energy lasers such as HeNe lasers are used to activate the photosensitized cellular target. A major problem associated with these studies is that events occurring during actual exposure of the target cells to the system cannot be examined in real time. In this study HeLa cells were photosensitized and photodynamic activation was accomplished using the scanning microbeam from a confocal laser scanning microscope. This form of activation allowed for simultaneous photoactivation and observation and facilitated the recording of events at a microscopic level during photoactivation. Effects of photodynamic activation on the target cells were monitored using the fluorophores rhodamine 123 and ethidium homodimer-1. Potential applications of these forms of analyses to space medicine and cell biology are discussed.     

DTFT-1: Analysis of the first USV flight test

The first dropped transonic flight test (DTFT) of the USV Program, performed with Castor, the first of the two spacecrafts developed within the USV Program, was performed on Saturday 24th February 2007, from Tortolì Airport in Sardinia.At 8:30 a.m. the 340 000 m³ stratospheric balloon lifted off from the East coast of Sardinia, bringing the flying test bed (FTB) up to 20.2 km before release within the isolated sea polygon controlled by Italian air force test range in Salto di Quirra (PISQ). The mission ended at 10:30 a.m. with the splash-down of the space vehicle.The flight itself was very good, with a nose-up manoeuvre under transonic conditions, reaching a maximum Mach as high as 1.08. The mission target was completely achieved as some 2 million measures were taken related to flight data, housekeeping, as well as 500 aerodynamic and structural experimental sensors. Unfortunately, the vehicle has been damaged more than expected during splash-down.Many national and international institutions and industries contributed to the mission carrying out, under the supervision and technical guide of CIRA: Italian Space Agency, Italian Air Force, Italian Navy, Italian Civil Aviation Authority, Italian Company for Air Navigation Services, Port Authorities, European Space Agency, Techno System Dev., Vitrociset, Carlo Gavazzi Space, Space Software Italia, Alcatel Alenia Space Italy, ISL-Altran Group.The paper reports the actual status of post-flight data analysis.