Search for life on Mars: evaluation of techniques.

An important question for exobiology is, did life evolve on Mars? To answer this question, experiments must be conducted on the martian surface. Given current mission constraints on mass, power, and volume, these experiments can only be performed using proposed analytical techniques such as: electron microscopy, X-ray fluorescence, X-ray diffraction, alpha-proton backscatter, gamma-ray spectrometry, differential thermal analysis, differential scanning calorimetry, pyrolysis gas chromatography, mass spectrometry, and specific element detectors. Using prepared test samples consisting of 1% organic matter (bovine serum albumin) in palagonite and a mixture of palagonite, clays, iron oxides, and evaporites, it was determined that a combination of X-ray diffraction and differential thermal analysis coupled with gas chromatography provides the best insight into the chemistry, mineralogy, and geological history of the samples.     

Origin of organic matter in the protosolar nebula and in comets.

Comet organics are traced to their origin in interstellar space. Possible sources of comet organics from solar nebula chemistry are briefly discussed. The infrared spectra of interstellar dust are compared with spectra of solar (space) irradiated laboratory organic residues and with meteorites. The spectra compare very favorably. The atomic composition of first generation laboratory organic residues compares favorably with that of comet Halley organics if divided into appropriate "volatile" (less refractory) and "refractory" (more refractory) complex organics.     

Did life exist on Mars? Search for organic and inorganic signatures, one of the goals for “SAM” (sample analysis at Mars)

Observation of Mars shows signs of a past Earth-like climate, and, in that case, there is no objection to the possible development of life, in the underground or at the surface, as in the terrestrial primitive biosphere. Sample analysis at Mars (SAM) is an experiment which may be proposed for atmospheric, ground and underground in situ measurements. One of its goals is to bring direct or indirect information on the possibility for life to have developed on Mars, and to detect traces of past or present biological activity. With this aim, it focuses on the detection of organic molecules: volatile organics are extracted from the sample by simple heating, whereas refractory molecules are made analyzable (i.e. volatile), using derivatization technique or fragmentation by pyrolysis. Gaseous mixtures thus obtained are analyzed by gas chromatography associated to mass spectrometry. Beyond organics, carbonates and other salts are associated to the dense and moist atmosphere necessary to the development of life, and might have formed and accumulated in some places on Mars. They represent another target for SAM. Heating of the samples allows the analysis of structural gases of these minerals (CO₂ from carbonates, etc.), enabling to identify them. We also show, in this paper, that it may be possible to discriminate between abiotic minerals, and minerals (shells, etc.) created by living organisms.     

The low temperature organic chemistry of Titan's geofluid.

Organic chemistry on Titan and prebiotic chemistry on Earth involve the same N-containing organics: nitriles and their oligomers. Couplings of their chemistry in the three parts of Titan's geofluid (atmosphere, aerosols and surface) seem to play a key role in the organic chemical evolution of the planet. If liquid water was present on Titan, then a prebiotic chemistry, involving eutectics, similar to that of the early Earth, may have occurred. However, liquid water is currently absent and a prebiotic chemistry based only on N-organics may be evolving now on Titan. The other consequence of the low temperatures of Titan is the possible formation of organics unstable at room temperature and very reactive. So far, these compounds have not been systematically searched for in experimental studies of Titan's organic chemistry. C4N2 has already been detected on Titan. Powerful reactants in organic chemistry, CH2N2, and CH3N3, may be also present. They exhibit spectral signatures in the mid-IR strong enough to allow their detection at the 10-100 ppb level. They may be detectable on future IR spectra (ISO and Cassini) of Titan.     

Closed and continuous algae cultivation system for food production and gas exchange in CELSS.

In CELSS (Controlled Ecological Life Support System), utilization of photosynthetic algae is an effective means for obtaining food and oxygen at the same time. We have chosen Spirulina, a blue-green alga, and have studied possibilities of algae utilization. We have developed an advanced algae cultivation system, which is able to produce algae continuously in a closed condition. Major features of the new system are as follows. (1) In order to maintain homogeneous culture conditions, the cultivator was designed so as to cause a swirl on medium circulation. (2) Oxygen gas separation and carbon dioxide supply are conducted by a newly designed membrane module. (3) Algae mass and medium are separated by a specially designed harvester. (4) Cultivation conditions, such as pH, temperature, algae growth rate, light intensity and quantity of generated oxygen gas are controlled by a computer system and the data are automatically recorded. This equipment is a primary model for ground experiments in order to obtain some design data for space use. A feasibility of algae cultivation in a closed condition is discussed on the basis of data obtained by use of this new system.     

The formation of organic molecules in astronomical ices.

An absorption feature at 3.4 micrometers has been observed in various lines-of-sight through the diffuse interstellar medium. Its position and width lead to an identification with the C-H stretching mode of solid organic material. A possible mechanism for the production of organic solids in the interstellar medium is UV photoprocessing of icy mantles which accrete on dust grains in dense clouds. Furthermore, thermally induced reactions involving formaldehyde molecules in the mantles could be an important source of organics. Laboratory simulation of these processes shows that a large variety of oxygen- and nitrogen-rich species may be produced. It is shown that the occurrence of periodic transient heating events plays an important role in the production of organic material in the ice mantles. Finally, it is pointed out how future missions like the Infrared Space Observatory (ISO) as well as analysis of comet material by Rosetta may be able to clarify the nature and evolution of interstellar organics.     

Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems.

Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. We report experiments in which cultures of the algae Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments we tested hydrolyzed waste biomass from these same algae to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.     

The electrolytical processes in dirty ices: implications for origin and chemistry of minor bodies and related objects.

Many moonlike bodies (M approximately or = 1 Moon) beyond the Martian orbit contain large amounts of dirty ice (approximately 50%) forming thick mantle with the solid phase thermal convection. When a body moves through the inter- or nearplanetary magnetized plasma, electric current is generated in the body and its environment. The current passing through a dirty ice containing up to 10% of organic admixtures produces a lot of electrochemical effects which have a profound impact on its composition. At this stage one can hardly say something definite concerning changes experienced by organics. The changes must occur inevitably and can be of a rather unexpected and far-reaching nature, so deserving a close study. Another obvious effect is a volumetric electrolysis of ice containing alien inclusions. The electrolysis products accumulate in ice in the form of a solid solution which is capable of detonation at 15-20 wt.% of 2H2 + O2. If M > or = 1 Moon (Galilean satellites, Titan), the body loses in explosion a part of its mass in the form of vapor and ice fragments (=short-period comet nuclei), whereas if M < or = 0.2 Moon, the body breaks up totally (the Main Belt asteroids origin approximately 3.9 Byr ago). 2H2 + O2 containing cometary nuclei are capable of burning or suffer new explosions when receiving an additional energy. The combustion in the sublimation products containing also light organics and 2H2 + O2 explains unexpected energetics and nearnuclear chemistry of Comet P/Halley (e.g. great abundances of negative and positive ions, atomic carbon, CO over CO2, origin of CHON particles etc) and its distant outbursts correlated, possibly, with the Solar activity. Thus the electrochemical processes in the dirty ice with organics, along with its subsequent thermal, radiative etc. processing, open up new potentials for explanation and prediction of quite unexpected discoveries.     

New insights into Titan's organic chemistry in the gas and aerosol phases.

Titan, the largest satellite of Saturn, with a dense atmosphere very rich in organics, and many couplings in the various parts of its "geofluid", is a reference for studying prebiotic chemistry on a planetary scale. New data have been obtained from experiments simulating this organic chemistry (gas and aerosol phases), within the right ranges of temperature and a careful avoiding of any chemical contamination. They show a very good agreement with the observational data, demonstrating for the first time the formation of all the organic species already detected in Titan atmosphere including, at last, C4N2, together with many other species not yet detected in Titan. This strongly suggests the presence of more complex organics in Titan's atmosphere and surface, including high molecular weight polyynes and cyanopolyynes. The NASA-ESA Cassini-Huygens mission has been successfully launched in October 1997. The Cassini spacecraft will reach the Saturn system in 2004 and become an orbiter around Saturn, while the Huygens probe will penetrate into Titan's atmosphere. In situ measurements, in particular from Huygens GC-MS and ACP instruments, will provide a detailed analysis of the organics present in the air, aerosols, and surface. This very ambitious mission should yield much information of crucial importance for our knowledge of the complexity of Titan's chemistry, and, more generally for the field of exobiology.     

Performance of a simple Closed Aquatic Ecosystem (CAES) in space.

A simple Closed Aquatic Ecosystem (CAES) consisting of single-celled green algae (Chlorella pyrenoidosa, producer), a spiral snail (Bulinus australianus, consumer) and a data acquisition and control unit was flown on the Chinese Spacecraft SHENZHOU-II in January 2001 for 7 days. In order to study the effect of microgravity on the operation of CAES, a 1 g centrifuge reference group in space, a ground 1 g reference group and a ground 1 g centrifuge reference group (1.4 g group) were run concurrently. Real-time data about algae biomass (calculated from transmission light intensity), temperature, light and centrifugation of the CAES were logged at minute intervals. It was found that algae biomass of both the microgravity group and the ground 1 g-centrifuge reference group (1.4 g) fluctuated during the experiment, but the algae biomass of the 1 g centrifuge reference group in space and the ground 1 g reference group increased during the experiment. The results may be attributable to influences of microgravity and 1.4 g gravity on the algae and snails metabolisms. Microgravity is the main factor to affect the operation of CAES in space and the contribution of microgravity to the effect was also estimated. These data may be valuable for the establishment of a complex CELSS in the future.     

Characterization of complex organic compounds formed in simulated planetary atmospheres by the action of high energy particles.

A wide variety of organic compounds, which are not simple organics but also complex organics, have been found in planets and comets. We reported that complex organics was formed in simulated planetary atmospheres by the action of high energy particles. Here we characterized the experimental products by using chromatographic and mass spectrometric techniques. A gaseous mixture of CO, N2 and H2O was irradiated with high energy protons (major components of cosmic rays). Water-soluble non-volatile substances, which gave amino acids after acid-hydrolysis, were characterized by HPLC and mass spectrometry. Major part of the products were complex compounds with molecular weight of several hundreds. Amino acid precursors were produced even when no water was incorporated with the starting materials. It was suggested that complex molecules including amino acid precursors were formed not in solution from simple molecules like HCN, but directly in gaseous phase.     

Operation of an experimental algal gas exchanger for use in a CELSS.

Concepts of a CELSS anticipate the use of photosynthetic organisms (higher plants and algae) for air revitalization. The rates of production and uptake of carbon dioxide and oxygen between the crew and the photosynthetic organisms are mismatched. An algal [correction of aglal] system used for gas exchange only will have the difficulty of an accumulation or depletion of these gases beyond physiologically tolerable limits (in a materially closed system the mismatch between assimilatory quotient (AQ) and respiratory quotient (RQ) will be balanced by the operation of the waste processor). We report the results of a study designed to test the feasibility of using environmental manipulations to maintain physiologically appropriate atmospheres for algae (Chlorella pyrenoidosa) and mice (Mus musculus strain DW/J) in a gas-closed system. Specifically, we consider the atmosphere behavior of this system with Chlorella grown on nitrate or urea and at different light intensities and optical densities. Manipulation of both the photosynthetic rate and AQ of the alga has been found to reduce the mismatch of gas requirements and allow operation of the system in a gas-stable manner. Operation of such a system in a CELSS may be useful for reduction of buffer sizes, as a backup system for higher plant air revitalization and to supply extra oxygen to the waste processor or during crew changes. In addition, mass balance for components of the system (mouse, algae and a waste processor) are presented.     

Design for a bioreactor with sunlight supply and operations systems for use in the space environment.

An experiment was carried out to determine the characteristics of an operations system that can support fast cultivation of algae at high densities in the weightlessness of space. The experiment was conducted in glass bioreactor tanks, in which light was supplied by radiator rods connected to optical fiber cables. The illumination areas of the tanks were 2600 cm2, 6000 cm2, and 9200 cm2 per liter of solution. The characteristics of O2-CO2 gas exchange, concentration and separation of chlorella in the growth medium, dialysis of ionic salts in the growth medium, etc. were examined. Chloralla ellipsoidea was used in the experiment, yielding the following results: (1) By increasing the ratio of illumination area to volume, growth rates of up to approximately 0.6 g/L h could be obtained in a highly concentrated solution (one that contains 20 g/L or more of algae). (2) The most suitable proportions of carbon dioxide and oxygen gases for growing algae quickly at high concentrations were found to be 10% CO2 and 10% O2 (by volume). (3) There was a high optimum concentration for fast cultivation, and the data obtained resembled the theoretical curve postulated by P. Behrens et al. (4) It was possible to exchange carbon dioxide and oxygen using gas-permeable membrane modules. (5) It was possible to separate the chlorella from the growth medium and recycle the medium.     

Exobiological exploration of Mars.

Of all the other planets in the solar system, Mars remains the most promising for further elucidating concepts about chemical evolution and the origin of life. Strategies were developed to pursue three exobiological objectives for Mars exploration: determining the abundance and distribution of the biogenic elements and organic compounds, detecting evidence of an ancient biota on Mars, and determining whether indigenous organisms exist anywhere on the planet. The three strategies are quite similar and, in fact, share the same sequence of phases. In the first phase, each requires global reconnaissance and remote sensing by orbiters to select sites of interest for detailed in situ analyses. In the second phase, lander missions are conducted to characterize the chemical and physical properties of the selected sites. The third phase involves conducting "critical" experiments at sites whose properties make them particularly attractive for exobiology. These critical experiments would include, for example, identification of organics, detection of fossils, and detection of extant life. The fourth phase is the detailed analysis of samples returned from these sites in Earth-based laboratories to confirm and extend previous discoveries. Finally, in the fifth phase, human exploration is needed to establish the geological settings for the earlier findings or to discover and explore sites that are not accessible to robotic spacecraft.     

Development of a ground-based space micro-algae photo-bioreactor

The purpose of the research is to develop a photo-bioreactor which may produce algae protein and oxygen for future astronauts in comparatively long-term exploration, and remove carbon dioxide in a controlled ecological life support system. Based on technical parameters and performance requirements, the project planning, design drafting, and manufacture were conducted. Finally, a demonstration test for producing algae was done. Its productivity for micro-algae and performance of the photo-bioreactor were evaluated. The facility has nine subsystems, including the reactor, the illuminating unit, the carbon dioxide (CO₂) production unit and oxygen (O₂) generation unit, etc. The demonstration results showed that the facility worked well, and the parameters, such as energy consumption, volume, and productivity for algae, met with the design requirement. The density of algae in the photo-bioreactor increased from 0.174 g (dry weight) L⁻¹ to 4.064 g (dry weight) L⁻¹ after 7 days growth. The principle of providing CO₂ in the photo-bioreactor for algae and removing O₂ from the culture medium was suitable for the demand of space conditions. The facility has reasonable technical indices, and smooth and dependable performances.     

The escape of molecular hydrogen and the synthesis of organic nitriles in planetary atmospheres.

What is the influence of hydrogen escape from the atmosphere of small planetary bodies on the synthesis of organic molecules in that atmosphere? To answer this question, laboratory experiments have been performed to study the evolution of different reducing model atmospheres submitted to electrical discharges, with and without the simulation of H₂ escape. A study of mixtures of nitrogen and methane shows a very strong effect of H₂ escape on the formation of organic nitriles, the only nitrogen containing organics detected in the gas phase. These are HCN, CH  CCN, (CN)₂, CH₂CHCN, CH₃ CN and CH₃CH₂CN. The yield of synthesis of most of these compounds is noticeably increased, up to several orders of magnitude, when hydrogen escape is simulated. The escape of H₂ from the atmosphere of the primitive Earth may have played a crucial role in the formation of reactive organic molecules such as CHCCN or (CN)₂, which can be considered as important prebiotic precursors. These experimental results may also explain extant data concerning the nature and relative abundance of organics present in the atmosphere of Titan, a planetary satellite which may be an ideal model within our solar system for the study of organic cosmochemistry and exobiology.     

Modelling of genetically engineered microorganisms introduction in closed artificial microcosms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E. coli decreased fast, and the structure of the plasmid was also modified. In conditions of algae blooming (Ankistrodesmus sp.) an almost total maintenance of plasmid-containing cells was observed in E. coli population. A mathematics model of GEM's behavior in water ecosystems with different level of complexity has been formulated. Mechanisms causing the difference in luminescent exhibition of different species are discussed, and attempts are made to forecast the GEM's behavior in water ecosystems.     

Thermally evolved gas analysis (TEGA) of hyperarid soils doped with microorganisms from the Atacama Desert in southern Peru: Implications for the Phoenix mission

TEGA, one of several instruments on board of the Phoenix Lander, performed differential scanning calorimetry and evolved gas analysis of soil samples and ice, collected from the surface and subsurface at a northern landing site on Mars. TEGA is a combination of a high temperature furnace and a mass spectrometer (MS) that was used to analyze samples delivered to the instrument via a robotic arm. The samples were heated at a programmed ramp rate up to 1000 °C. The power required for heating can be carefully and continuously monitored (scanning calorimetry). The evolved gases generated during the process can be analyzed with the evolved gas analyzer (a magnetic sector mass spectrometer) in order to determine the composition of gases released as a function of temperature. Our laboratory has developed a sample characterization method using a pyrolyzer integrated to a quadrupole mass spectrometer to support the interpretations of TEGA data. Here we examine the evolved gas properties of six types of hyperarid soils from the Pampas de La Joya in southern Peru (a possible analog to Mars), to which we have added with microorganisms (Salmonella typhimurium, Micrococcus luteus, and Candida albicans) in order to investigate the effect of the soil matrix on the TEGA response. Between 20 and 40 mg of soil, with or without ∼5 mg of lyophilized microorganism biomass (dry weight), were placed in the pyrolyzer and heated from room temperature to 1200 °C in 1 h at a heating rate of 20 °C/min. The volatiles released were transferred to a MS using helium as a carrier gas. The quadrupole MS was ran in scan mode from 10 to 200 m/z. In addition, ∼20 mg of each microorganism without a soil matrix were analyzed. As expected, there were significant differences in the gases released from microorganism samples with or without a soil matrix, under similar heating conditions. Furthermore, samples from the most arid environments had significant differences compared with less arid soils. Organic carbon released in the form of CO₂ (ion 44 m/z) from microorganisms evolved at temperatures of ∼326.0 ± 19.5 °C, showing characteristic patterns for each one. Others ions such as 41, 78 and 91 m/z were also found. Interestingly, during the thermal process, the release of CO₂ increased and ions previously found disappeared, demonstrating a high-oxidant activity in the soil matrix when it was subjected to high temperature. Finally, samples of soil show CO₂ evolved up to 650 °C consistent with thermal decomposition of carbonates. These results indicate that organics mixed with these hyperarid soils are oxidized to CO₂. Our results suggest the existence of at least two types of oxidants in these soils, a thermolabile oxidant which is highly oxidative and other thermostable oxidant which has a minor oxidative activity and that survives the heat-treatment. Furthermore, we find that the interaction of biomass added to soil samples gives a different set of breakdown gases than organics resident in the soil. The nature of oxidant(s) present in the soils from Pampas de La Joya is still unknown.     

A simple closed aquatic ecosystem (CAES) for space

A closed aquatic ecosystem (CAES) was developed to study the effects of microgravity on the function of closed ecosystems aboard the Chinese retrieved satellite and on the spacecraft SHENZHOU-II. These systems housed a small freshwater snail (Bulinus australianus) and an autotrophic green algae (Chlorella pyrenoidosa). The results of the test on the satellite were that the concentration of algae changed little, but that the snails died during the experiments. We then sought to optimize the function of the control system, the cultural conditions and the data acquisition system and carried out an experiment on the spacecraft SHENZHOU-II. Using various sensors to monitor the CAES, real-time data regarding the operation of the CAES in microgravity was acquired. In addition, an on-board 1g centrifuge was included to identify gravity-related factors. It was found that microgravity is the major factor affecting the operation of the CAES in space. The change in biomass of the primary producer during each day in microgravity was larger than that of the control groups. The mean biomass concentration per day in the microgravity group decreased, but that of the control groups increased for several days and then leveled off. Space effects on the biomass of a primary producer may be a result of microgravity effects leading to increasing metabolic rates of the consumer combined with decreases in photosynthesis.     

Possible complex organic compounds on Mars.

It is suggested that primitive Mars had somehow similar environments as primitive Earth. If life was born on the primitive earth using organic compounds which were produced from the early Earth environment, the same types of organic compounds were also formed on primitive Mars. Such organic compounds might have been preserved on Mars still now. We are studying possible organic formation on primitive and present Mars. A gaseous mixture of CO2, CO, N2 and H2O with various mixing ratios were irradiated with high energy protons (major components of cosmic rays). Hydrogen cyanide and formaldehyde were detected among volatile products, and yellow-brown-colored water-soluble non-volatile substances were produced, which gave amino acids after acid-hydrolysis. Major part of "amino acid precursors" were not simple molecules like aminonitriles, but complex compounds which eluted earlier than free amino acids in cation-exchange HPLC. These organic compounds should be major targets in the future Mars mission. Strategy for the detection of the complex organics on Mars will be discussed.