Exobiology and future Mars missions: The search for Mars'' earliest biosphere

The primordial Mars may have possessed a thick carbon dioxide atmosphere, with liquid water common on the surface, similar in many ways to the primordial Earth. During this epoch, billions of years ago, the surface of Mars could have been conducive to the origin of life. It is possible that life evolved on Mars to be later eliminated as the atmospheric pressure dropped. Analysis of the surface of Mars for the traces of this early martian biota could provide many insights into the phenomenon of life and its coupling to planetary evolution.     

The use of mineral crystals as bio-markers in the search for life on Mars.

Photographs that depict presumed fluvial features on the martian surface have led geologists to hypothesize that water flowed across the early martian terrain. From this, it has been further hypothesized that the surface and atmospheric conditions on early Mars were similar to those on early Earth. Because the oldest fossil evidence of life on Earth dates back to this early period, at least 3.5 billion years ago, the possibility exists that the early Martian environment could have also been conducive to the origin of life. To investigate this possibility, universal signatures or bio-markers indicative of past (or present) biological activity must be identified for use in the search for life on Mars. Several potentially applicable biomarkers have been identified and include: organics (e.g., specific classes of lipids and hopanes), suites of specific inorganic and organic compounds, as well as the isotopic ratios of C, N, and S. Unfortunately, all of these bio-markers may be of biologic or abiotic origin; these origins are often difficult to distinguish. Thus, the discovery of any one of these compounds alone is not a bio-marker. Because minerals produced under biologic control have distinctive crystallographies, morphologies, and isotopic ratios that distinguishable from abiotically produced minerals with the same chemical composition, and are stable through geologic time, we propose the use of minerals resulting from biologically controlled mineralization processes as bio-markers.     

Life on Mars? II. Physical restrictions.

The primary physical factors important to life's evolution on a planet include its temperature, pressure and radiation regimes. Temperature and pressure regulate the presence and duration of liquid water on the surface of Mars. The prolonged presence of liquid water is essential for the evolution and sustained presence of life on a planet. It has been postulated that Mars has always been a cold dry planet; it has also been postulated that early mars possessed a dense atmosphere of CO2 (> or = 1 bar) and sufficient water to cut large channels across its surface. The degree to which either of these postulates is true correlates with the suitability of Mars for life's evolution. Although radiation can destroy living systems, the high fluxes of UV radiation on the martian surface do not necessarily stop the origin and early evolution of life. The probability for life to have arisen and evolved to a significant degree on Mars, based on the postulated ranges of early martian physical factors, is almost solely related to the probability of liquid water existing on the planet for at least hundreds of millions to billions of years.     

A preliminary comparison of two perennially ice-covered lakes in Antarctica: analogs of past Martian lacustrine environments.

Perennially ice-covered lakes in the Antarctic have been suggested as analogs to lakes which may have existed on the surface of Mars 3.5 billion years ago. During the 1991-1992 austral summer, a joint Russian/American research effort was directed at studies of ice-covered lakes in the Bunger Hills Oasis, Antarctica (66 degrees S, 100 degrees E). The primary objective of the expedition was to investigate this ice-free area for features analogous to ancient martian environments that may have been capable of supporting life and to compare the ice-covered lakes of the Bunger Hills with those in the McMurdo Dry Valleys of southern Victoria Land (77 degrees S, 166 degrees E) as part of the continuing studies of Antarctic-Mars analogs.     

Planetary Protection Policy (U.S.A.).

Through existing treaty obligations of the United States, NASA is committed to exploring space while avoiding biological contamination of the planets, and to the protection of the Earth against harm from materials returned from space. Because of the similarities between Mars and Earth, plans for the exploration of Mars evoke discussions of these Planetary Protection issues. US Planetary Protection Policy will be focused on the preservation of these goals in an arena that will change with the growth of scientific knowledge about the martian environment. Early opportunities to gain the appropriate data will be used to guide later policy implementation. Because human presence on Mars will result in the end of Earth's separation from the martian environment, it is expected that precursor robotic missions will address critical planetary protection concerns before humans arrive.     

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.     

Bio-markers and the search for extinct life on Mars.

Geologic and climatologic studies suggest that conditions on early Mars were similar to early Earth. Because life on Earth is believed to have originated during this early period (3.5 billion years ago), the Martian environment could have also been conducive to the origin of life. To investigate this possibility we must first define the attributes of an early Martian biota. Then, specific geographic locations on Mars must be chosen where life may have occurred (i.e. areas which had long standing water), and within these distinct locations search for key signatures or bio-markers of a possible extinct Martian biota. Some of the key signatures or bio-markers indicative of past biological activity on Earth may be applicable to Mars including: reduced carbon and nitrogen compounds, CO3(2-), SO4(2-), NO3-, NO2- [correction of NO2(2)], Mg, Mn, Fe, and certain other metals, and the isotopic ratios of C, N and S. However, we must also be able to distinguish abiotic from biologic origins for these bio-markers. For example, abiotically fixed N2 would form deposits of NO3- and NO2-, whereas biological processes would have reduced these to ammonium containing compounds, N2O, or N2, which would then be released to the atmosphere. A fully equipped Mars Rover might be able to perform analyses to measure most of these biomarkers while on the Martian surface.     

Selection of sterilization methods for planetary return missions.

Two tasks must be accomplished to provide planetary protection for Mars return missions: (1) sterilization of the scientific module to be landed on Mars and (2) reliable sterilization of all material returned to Earth, while ensuring the scientific integrity of martian samples. This paper examines similarity and differences between these two tasks, and includes a discussion of technological implementation conditions and the nature of terrestrial and hypothesized martian microflora. The feasibility of a number of chemical and physical (ultraviolet and ionizing radiation and heating) methods of sterilization for use on the ground and onboard are discussed and compared. A combination of different methods will probably be selected as the most appropriate for ensuring planetary protection on the return mission.     

Regenerative life support systems--why do we need them?

Human exploration of the solar system will include missions lasting years at a time. Such missions mandate extensive regeneration of life support consumables with efficient utilization of local planetary resources. As mission durations extend beyond one or two years, regenerable human life support systems which supply food and recycle air, water, and wastes become feasible; resupply of large volumes and masses of food, water, and atmospheric gases become unrealistic. Additionally, reduced dependency on resupply or self sufficiency can be an added benefit to human crews in hostile environments far from the security of Earth. Comparisons of resupply and regeneration will be discussed along with possible scenarios for developing and implementing human life support systems on the Moon and Mars.     

Detection of martian amino acids by chemical derivatization coupled to gas chromatography: in situ and laboratory analysis.

If there is, or ever was, life in our solar system beyond the Earth, Mars is the most likely place to search for. Future space missions will have then to take into account the detection of prebiotic molecules or molecules of biological significance such as amino acids. Techniques of analysis used for returned samples have to be very sensitive and avoid any chemical or biological contamination whereas in situ techniques have to be automated, fast and low energy consuming. Several possible methods could be used for in situ amino acid analyses on Mars, but gas chromatography would likely be the most suitable. Returned samples could be analyzed by any method in routine laboratory use such as gas chromatography, already successfully performed for analyses of organic matter including amino acids from martian meteorites. The derivatization step, which volatilizes amino acids to perform both in situ and laboratory analysis by gas chromatography, is discussed here.     

Science strategy for human exploration of Mars.

The scientific objectives of Mars exploration can be framed within the overarching theme of exploring Mars as another home for life, both for evidence of past or present life on Mars, and as a potential future home for human life. The two major areas of research within this theme are: 1) determining the relationship between planetary evolution, climate change, and life, and 2) determining the habitability of Mars. Within this framework, this paper discusses the exploration objectives for exobiology, climatology and atmospheric science, geology, and martian resource assessment. Human exploration will proceed in four major phases: 1) Precursor missions which will obtain environmental knowledge necessary for human exploration, 2) Emplacement phase which includes the first few human landings where crews will explore the local area of the landing site; 3) Consolidation phase missions where a permanent base will be constructed and crews will be capable of detailed exploration over regional scales; 4) Utilization phase, in which a continuously occupied permanent Mars base exists and humans will be capable of detailed global exploration of the martian surface. The phases of exploration differ primarily in the range and capabilities of human mobility. In the emplacement phase, an unpressurized rover, similar to the Apollo lunar rover, will be used and will have a range of a few tens of kilometers. In the Consolidation phase, mobility will be via a pressurized all-terrain vehicle capable of expeditions from the base site of several weeks duration. In the Utilization phase, humans will be capable of several months long expeditions to any point on the surface of Mars using a suborbital rocket equipped with habitat, lab, and return vehicle. Because of human mobility limitations, it is important to extend the range and duration of exploration in all phases by using teleoperated rover vehicles. Site selection for human missions to Mars must consider the multi-decade time frame of these four phases. We suggest that operations in the first two phases be focused in the regional area containing the Coprates Quadrangle and adjacent areas.     

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen.

Early Earth and early Mars were similar enough such that past geochemical and climatic conditions on Mars may have also been favorable for the origin of life. However, one of the most striking differences between the two planets was the low partial pressure of dinitrogen (pN2) on early Mars (18 mb). On Earth, nitrogen is a key biological element and in many ecosystems the low availability of fixed nitrogen compounds is the main factor limiting growth. Biological fixation of dinitrogen on Earth is a crucial source of fixed nitrogen. Could the low availability of dinitrogen in the primordial Martian atmosphere have prevented the existence, or evolution of Martian microbiota? Azotobacter vinelandii and Azomonas agilis were grown in nitrogen free synthetic medium under various partial pressures of dinitrogen ranging from 780-0 mb (total atmosphere=1 bar). Below 400 mb the biomass, cell number, and growth rate decreased with decreasing pN2. Both microorganisms were capable of growth at a pN2 as low as 5 mb, but no growth was observed at a pN2 < or = 1 mb. The data appear to indicate that biological nitrogen fixation could have occurred on primordial Mars (pN2=18 mb) making it possible for a biotic system to have played a role in the Martian nitrogen cycle. It is possible that nitrogen may have played a key role in the early evolution of life on Mars, and that later a lack of available nitrogen on that planet (currently, pN2=0.2 mb) may have been involved in its subsequent extinction.     

Simulation of the environmental climate conditions on martian surface and its effect on Deinococcus radiodurans

The resistance of terrestrial microorganisms under the thermo-physical conditions of Mars (diurnal temperature variations, UV climate, atmospheric pressure and gas composition) at mid-latitudes was studied for the understanding and assessment of potential life processes on Mars. In order to accomplish a targeted search for life on other planets, e.g. Mars, it is necessary to know the limiting physical and chemical parameters of terrestrial life. Therefore the polyextremophile bacterium Deinococcus radiodurans was chosen as test organism for these investigations. For the simulation studies at the Planetary and Space Simulation Facilities (PSI) at DLR, Cologne, Germany, conditions that are present during the southern summer at latitude of 60° on Mars were applied.We could simulate several environmental parameters of Mars in one single experiment: vacuum/low pressure, anoxic atmosphere and diurnal cycles in temperature and relative humidity, energy-rich ultraviolet (UV) radiation as well as shielding by different martian soil analogue materials. These parameters have been applied both single and in different combinations in laboratory experiments. Astonishingly the diurnal Mars-like cycles in temperature and relative humidity affected the viability of D. radiodurans cells quite severely. But the martian UV climate turned out to be the most deleterious factor, though D. radiodurans is red-pigmented due to carotenoids incorporated in its cell wall, which have been assigned not only a possible role as free radical scavenger but also as a UV-protectant. An additional UV-protection was accomplished by mixing the bacteria with nano-sized hematite.     

Survival and death of the haloarchaeon Natronorubrum strain HG-1 in a simulated martian environment

Halophilic archaea are of interest to astrobiology due to their survival capabilities in desiccated and high salt environments. The detection of remnants of salty pools on Mars stimulated investigations into the response of haloarchaea to martian conditions. Natronorubrum sp. strain HG-1 is an extremely halophilic archaeon with unusual metabolic pathways, growing on acetate and stimulated by tetrathionate. We exposed Natronorubrum strain HG-1 to ultraviolet (UV) radiation, similar to levels currently prevalent on Mars. In addition, the effects of low temperature (4, −20, and −80 °C), desiccation, and exposure to a Mars soil analogue from the Atacama desert on the viability of Natronorubrum strain HG-1 cultures were investigated. The results show that Natronorubrum strain HG-1 cannot survive for more than several hours when exposed to UV radiation equivalent to that at the martian equator. Even when protected from UV radiation, viability is impaired by a combination of desiccation and low temperature. Desiccating Natronorubrum strain HG-1 cells when mixed with a Mars soil analogue impaired growth of the culture to below the detection limit. Overall, we conclude that Natronorubrum strain HG-1 cannot survive the environment currently present on Mars. Since other halophilic microorganisms were reported to survive simulated martian conditions, our results imply that survival capabilities are not necessarily shared between phylogenetically related species.     

Planetary protection program for Mars 94/96 mission.

Mars surface in-situ exploration started in 1975 with the American VIKING mission. Two probes landed on the northern hemisphere and provided, for the first time, detailed information on the martian terrain, atmosphere and meteorology. The current goal is to undertake larger surface investigations and many projects are being planned by the major Space Agencies with this objective. Among these projects, the Mars 94/96 mission will make a major contributor toward generating significant information about the martian surface on a large scale. Since the beginning of the Solar System exploration, planets where life could exist have been subject to planetary protection requirements. Those requirements accord with the COSPAR Policy and have two main goals: the protection of the planetary environment from influence or contamination by terrestrial microorganisms, the protection of life science, and particularly of life detection experiments searching extra-terrestrial life, and not life carried by probes and spacecrafts. As the conditions for life and survival for terrestrial microorganisms in the Mars environment became known, COSPAR recommendations were updated. This paper will describe the decontamination requirements which will be applied for the MARS 94/96 mission, the techniques and the procedures which are and will be used to realize and control the decontamination of probes and spacecrafts.     

The search for nitrogen compounds on the surface of Mars

Nitrogen is an essential element for life. Specifically, “fixed nitrogen” (i.e., NH₃, NH₄⁺, NO_x, or N that is chemically bound to either inorganic or organic molecules and is releasable by hydrolysis to NH₃ or NH₄⁺) is the form of nitrogen useful to living organisms. To date no direct analysis of Martian soil nitrogen content, or content of fixed nitrogen compounds has been done. Consequently, the planet's total inventory of nitrogen is unknown. What is known is that the N₂ content of the present-day Martian atmosphere is 0.2 mbar. It has been hypothesized that early in Mars' history (3 to 4 billion years ago) the Martian atmosphere contained much more N₂ than it does today. The values of N₂ proposed for this early Martian atmosphere, however, are not well constrained and range from 3 to 300 mbar of N₂. If the early atmosphere of Mars did contain much more N₂ than it does today the question to be answered is, Where did it go? The two main processes that could have removed it rapidly from the atmosphere include: 1) nonthermal escape of N-atoms to space; and 2) burial within the regolith as nitrates and nitrites. Nitrate will be stable in the highly oxidized surface soil of Mars, and will tend to accumulate in the soil. Such accumulations are observed in certain desert environments on Earth. Some NH₄⁺---N may also be fixed and stabilized in the soil by inclusion as a structural cation in the crystal lattices of certain phyllosilicates replacing K. Analysis of the Martian soil for traces of NO₃⁻ and NH₄⁺ during future missions will supply important information regarding the nitrogen abundance on Mars, its past climate as well as its potential for the evolution of life.     

Planetary protection issues and the future exploration of Mars.

A primary scientific theme for the Space Exploration Initiative (SEI) is the search for life, extant or extinct, on Mars. Because of this, concerns about Planetary Protection (PP), the prevention of biological cross-contamination between Earth and other planets during solar system exploration missions, have arisen. A recent workshop assessed the necessity for, and impact of, PP requirements on the unmanned and human missions to Mars comprising the SEI. The following ground-rules were adopted: 1) information needed for assessing PP issues must be obtained during the unmanned precursor mission phase prior to human landings; 2) returned Mars samples will be considered biologically hazardous until proven otherwise; 3) deposition of microbes on Mars and exposure of the crew to Martian materials are inevitable when humans land; and, 4) human landings are unlikely until it is demonstrated that there is no harmful effect of Martian materials on terrestrial life forms. These ground-rules dictated the development of a conservative PP strategy for precursor missions. Key features of the proposed strategy include: 1) for prevention of forward contamination, all orbiters will follow Mars Observer PP procedures for assembly, trajectory, and lifetime. All landers will follow Viking PP procedures for assembly, microbial load reduction, and bioshield; and, 2) for prevention of back contamination, all sample return missions will have PP requirements which include fail-safe sample sealing, breaking contact chain with the Martian surface, and containment and quarantine analysis in an Earth-based lab. In addition to deliberating on scientific and technical issues, the workshop made several recommendations for dealing with forward and back contamination concerns from non-scientific perspectives.     

Life on Mars? I. The chemical environment.

The origin of life at its abiotic evolutionary stage, requires a combination of constituents and environmental conditions that enable the synthesis of complex replicating macromolecules from simpler monomeric molecules. It is very likely that the early stages of this evolutionary process have been spontaneous, rapid and widespread on the surface of the primitive Earth, resulting in the formation of quite sophisticated living organisms within less than a billion years. To what extent did such conditions prevail on Mars? Two companion-papers (Life on Mars? I and II) will review and discuss the available information related to the chemical, physical and environmental conditions on Mars and assess it from the perspective of potential exobiological evolution.     

Life in the solar system.

Life, defined as a chemical system capable of transferring its molecular information via self-replication and also capable of evolving, must develop within a liquid to take advantage of the diffusion of complex molecules. On Earth, life probably originated from the evolution of reduced organic molecules in liquid water. Organic matter might have been formed in the primitive Earth's atmosphere or near hydrothermal vents. A large fraction of prebiotic organic molecules might have been brought by extraterrestrial-meteoritic and cometary dust grains decelerated by the atmosphere. Any celestial body harboring permanent liquid water may therefore accumulate the ingredients that generated life on the primitive Earth. The possibility that life might have evolved on early Mars when water existed on the surface marks it as a prime candidate in a search for bacterial life beyond the Earth. Europa has an icy carapace. However, cryovolcanic flows at the surface point to a possible water subsurface region which might harbor a basic life form. The atmosphere and surface components of Titan are also of interest to exobiology for insight into a hydrocarbon-rich chemically evolving world. One-handed complex molecules and preferential isotopic fractionation of carbon, common to all terrestrial life forms, can be used as basic indicators when searching for life beyond the Earth.