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.     

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.     

Studies in the search for life on Mars.

The ability of living organisms to survive extraterrestrial conditions has implications for the origins of life in the solar system. We have therefore studied the survival of viruses, bacteria, yeast, and fungi under simulated Martian conditions. The environment on Mars was simulated by low temperature, proton irradiation, ultraviolet irradiation, and simulated Martian atmosphere (CO2 95.46%, N2 2.7%, water vapor 0.03%) in a special cryostat. After exposure to these conditions, tobacco mosaic virus and spores of Bacillus, Aspergillus, Clostridium, and some species of coccus showed significant survival.     

Stable carbon isotopes: possible clues to early life on Mars.

Organic and inorganic carbon in terrestrial near-surface environments are characterized by a marked difference in their 13C/12C ratios which can be traced back in the Earth's sedimentary record over almost 4 billion years. There is no doubt that the bias in favour of 12C displayed by biogenic matter derives, for the most part, from the isotope-selecting properties of the carbon-fixing enzyme (ribulose-1,5-bisphosphate carboxylase) that is operative in the principal photosynthetic pathway and promotes most of the carbon transfer from the non-living to the living realm. Postulating a universality of biological principles in analogy to the proven universality of the laws of physics and chemistry, we may expect enzymatic reactions in exobiological systems to be beset with B similar kinetic fractionation effects. Hence, the retrieval from the oldest Martian sediments of isotopic fractionations between reduced and oxidized (carbonate) carbon may substantially constrain current conjectures on the possible existence of former life on Mars.     

Chemical studies on the possible existence of life on Mars.

Although there is no direct evidence yet for the existence of life on Mars, it is reasonable to conclude that the emergence of life on Earth, which appears to have been controlled by universal laws of physics and chemistry, may have been repeated elsewhere in the universe. The dual approach of synthesis and analysis in our experimental studies has provided ample evidence in support of this hypothesis.     

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.     

The antarctic cold desert and the search for traces of life on Mars

The cryptoendolithic microorganisms that live inside rocks in the frigid Ross Desert of Antarctica can serve as a terrestrial model for what may have happened to life forms on Mars when the planet became dry and cold. Trace fossils of microbial rock colonization exist in Antarctica, and similar structures could ave formed on Mars. In some respects, such trace fossils could be an easier target for life-detection systems than fossils of cellular structures.     

In-situ resource utilization technologies for Mars life support systems.

The atmosphere of Mars has many of the ingredients that can be used to support human exploration missions. It can be "mined" and processed to produce oxygen, buffer gas, and water, resulting in significant savings on mission costs. The use of local materials, called ISRU (for in-situ resource utilization), is clearly an essential strategy for a long-term human presence on Mars from the standpoints of self-sufficiency, safety, and cost. Currently a substantial effort is underway by NASA to develop technologies and designs of chemical plants to make propellants from the Martian atmosphere. Consumables for life support, such as oxygen and water, will probably benefit greatly from this ISRU technology development for propellant production. However, the buffer gas needed to dilute oxygen for breathing is not a product of a propellant production plant. The buffer gas needs on each human Mars mission will probably be in the order of metric tons, primarily due to losses during airlock activity. Buffer gas can be separated, compressed, and purified from the Mars atmosphere. This paper discusses the buffer gas needs for a human mission to Mars and consider architectures for the generation of buffer gas including an option that integrates it to the propellant production plant.     

Stable carbon isotope fractionation in the search for life on early Mars.

Isotopic measurements and, more specifically, ratios of 13C to 12C in organic relative to inorganic deposits, are useful in reconstructing past biological activity on Earth. Organic matter has a lower ratio of 13C to 12C due largely to the preferential fixation of 12C over the heavier isotope by the major carbon-fixation enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase, although other factors (e.g., availability of source carbon, fixation by other carboxylating enzymes and diagenesis of organic material) also contribute to fractionation. Would carbon isotope discrepancies between inorganic and organic carbon indicate past biological activity on Mars? In order to answer this question, we analyse what is known about terrestrial biologic and abiologic carbon fixation and its preservation in the fossil record, and suggest what the isotope discrimination during possible biologic and abiologic carbon fixation on Mars might have been like. Primarily because isotopic signatures of abiotically fixed carbon overlap with those of biotic fixation, but also because heterotrophy does not significantly alter the isotopic signature of ingested carbon, fractionation alone would not be definitive evidence for life. However, a narrow range of fractionation, including no fractionation, would suggest biotic processes. Never-the-less, isotopic ratios in organic deposits on Mars would be extremely useful in analysing prebiotic, if not biotic, carbon transformations on Mars.     

Planetary protection and the search for life beneath the surface of Mars.

The search for traces of extinct and extant life on Mars will be extended to beneath the surface of the planet. Current data from Mars missions suggesting the presence of liquid water early in Mars' history and mathematical modeling of the fate of water on Mars imply that liquid water may exist deep beneath the surface of Mars. This leads to the hypothesis that life may exist deep beneath the Martian surface. One possible scenario to look for life on Mars involves a series of unmanned missions culminating with a manned mission drilling deep into the Martian subsurface (approximately 3Km), collecting samples, and conducting preliminary analyses to select samples for return to earth. This mission must address both forward and back contamination issues, and falls under planetary protection category V. Planetary protection issues to be addressed include provisions stating that the inevitable deposition of earth microbes by humans should be minimized and localized, and that earth microbes and organic material must not contaminate the Martian subsurface. This requires that the drilling equipment be sterilized prior to use. Further, the collection, containment and retrieval of the sample must be conducted such that the crew is protected and that any materials returning to earth are contained (i.e., physically and biologically isolated) and the chain of connection with Mars is broken.     

A "cryptic" microbial mat: a new model ecosystem for extant life on Mars.

If life were present on Mars to day, it would face potentially lethal environmental conditions such as a lack of water, frigid temperatures, ultraviolet radiation, and soil oxidants. In addition, the Viking missions did not detect near-surface organic carbon available for assimilation. Autotrophic organisms that lived under a protective layer of sand or gravel would be able to circumvent the ultraviolet radiation and lack of fixed carbon. Two terrestrial photosynthetic near-surface microbial communities have been identified, one in the inter- and supertidal of Laguna Ojo de Liebre (Baja California Sur, Mexico) and one in the acidic gravel near several small geysers in Yellowstone National Park (Wyoming, U.S.A.). Both communities have been studied with respect to their ability to fix carbon under different conditions, including elevated levels of inorganic carbon. Although these sand communities have not been exposed to the entire suite of Martian environmental conditions simultaneously, such communities can provide a useful model ecosystem for a potential extant Martian biota.     

Phosphorus as a potential guide in the search for extinct life on Mars.

In contrast to the search for extant organisms, the quest for fossil remains of life on Mars need not be guided by the presence of water and organic compounds on the present surface. An appropriate tracer might be the element phosphorus which is a common constituent of living systems. Utilizing terrestrial analogues, it should preferentially exist in the form of sedimentary calcium phosphate (phosphorites), which would have readily resisted changing conditions on Mars. Moreover, higher ratios of P/Th in phosphorites in comparison to calcium phosphates from magmatic rocks give us the possibility to distinguish them from inorganically formed phosphorus deposits at or close to the Martian surface. Identification of anomalous phosphorus enrichments by remote sensing or in situ analysis could be promising approaches for selecting areas preferentially composed of rocks with remains of extinct life.     

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.     

Biosphere 2: a prototype project for a permanent and evolving life system for Mars base.

As part of the ground-based preparation for creating long-term life systems needed for space habitation and settlement, Space Biospheres Ventures (SBV) is undertaking the Biosphere 2 project near Oracle, Arizona. Biosphere 2, currently under construction, is scheduled to commence its operations in 1991 with a two-year closure period with a crew of eight people. Biosphere 2 is a facility which will be essentialy materially-closed to exchange with the outside environment. It is open to information and energy flow. Biosphere 2 is designed to achieve a complex life-support system by the integration of seven areas or "biomes"--rainforest, savannah, desert, marsh, ocean, intensive agriculture and human habitat. Unique bioregenerative technologies, such as soil bed reactors for air purification, aquatic waste processing systems, real-time analytic systems and complex computer monitoring and control systems are being developed for the Biosphere 2 project. Its operation should afford valuable insight into the functioning of complex life systems necessary for long-term habitation in space. It will serve as an experimental ground-based prototype and testbed for the stable, permanent life systems needed for human exploration of Mars.     

History of water on Mars: a biological perspective.

We divide the history of water on the Martian surface into four epochs based upon the atmospheric temperature and pressure. In Epoch 1, during which a primordial CO2 atmosphere was actively maintained by impact and volcanic recycling, we presume the mean annual temperature to have been above freezing, the pressure to have exceeded one atmosphere, and liquid water to have been widespread. Under such conditions, similar to early Earth, life could have arisen and become abundant. After this initial period of recycling, atmospheric CO2 was irreversibly lost due to carbonate formation and the pressure and temperature declined. In Epoch II, the mean annual temperature fell below freezing but peak temperatures would have exceeded freezing. Ice covered lakes, similar to those in the McMurdo Dry Valleys of Antarctica could have provided a habitat for life. In Epoch III, the mean and peak temperatures were below freezing and there would have been only transient liquid water. Microbial ecosystems living in endolithic rock "greenhouses" could have continued to survive. Finally, in Epoch IV, the pressure dropped to near the triple point pressure of water and liquid water could no longer have existed on the surface and life on the surface would have become extinct.     

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.     

Remote sensing for organics on Mars.

The detection of organics on Mars remains an important scientific objective. Advances in instrumentation and laboratory techniques provide new insight into the lower level detection limit of complex organics in closely packed media. Preliminary results demonstrate that algae present in a palagonite medium do exhibit a spectral reflectance feature in the visible range for dry mass weight ratios of algae to palagonite greater than 6%--which corresponds to 30 mg algae in a 470 mg (just optically thick (< 3 mm) layer) palagonite matrix. This signature most probably represents chlorophyll a, a light harvesting pigment with an emission peak at 678 nm.     

Biogeochemical evidence of microbial activity on Mars.

We suggest a new interpretation of the data on so-called SNC meteorites and delta 13C values of the calcium carbonate minerals and organic matter discovered in them. The delta 13C value of calcite (up to 15 ppt) is accounted for by the microbial reaction CO2 + H2 ---> CH4 + H2O. Methane-forming bacteria also synthesize organic carbon (in the form of biomass) from CO2, and this process is accompanied by 12C fractionation. Therefore, the organic carbon of SNC meteorites is enriched with 12C (delta 13C as low as -35 ppt). The environmental conditions under which the calcite of SNC meteorites was formed were favorable for the activity of methanogens.     

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.     

Estimation and assessment of Mars contamination.

Since the beginning of the exploration of Mars, more than fourty years ago, thirty-six missions have been launched, including fifty-nine different space systems such as fly-by spacecraft, orbiters, cruise modules, landing or penetrating systems. Taking into account failures at launch, about three missions out of four have been successfully sent toward the Red Planet. The fact today is that Mars orbital environment includes orbiters and perhaps debris, and that its atmosphere and its surface include terrestrial compounds and dormant microorganisms. Coming from the UN Outer Space Treaty [United Nations Treaty on Principles Governing the Activities of States in the Exploration and Use of Outer Space, including the Moon and Other Celestial Bodies (the "Outer Space Treaty") referenced 610 UNTS 205 - resolution 2222(XXI) of December 1966] and according to the COSPAR planetary protection policy recommendations [COSPAR Planetary Protection Policy (20 October 2002), accepted by the Council and Bureau, as moved for adoption by SC F and PPP, prepared by the COSPAR/IAU Workshop on Planetary Protection, 4/02 with updates 10/0, 2002], Mars environment has to be preserved so as not to jeopardize the scientific investigations, and the level of terrestrial material brought on and around Mars theoretically has to comply with this policy. It is useful to evaluate what and how many materials, compounds and microorganisms are on Mars, to list what is in orbit and to identify where all these items are. Considering assumptions about materials, spores and gas location and dispersion on Mars, average contamination levels can be estimated. It is clear now that as long as missions are sent to other extraterrestrial bodies, it is not possible to keep them perfectly clean. Mars is one of the most concerned body, and the large number of missions achieved, on-going and planned now raise the question about its possible contamination, not necessarily from a biological point of view, but with respect to all types of contamination. Answering this question, will help to assess the potential effects of such contamination on scientific results and will address concerns relative to any ethical considerations about the contamination of other planets.