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Non-Equilibrium Thermodynamic Chemistry and the Composition of the Atmosphere of Mars

A high priority objective of the Mars Exploration Program is to Determine if life exists today (MEPAG Goal I, Objective A). The measurement of gases of biogenic origin may be an approach to detect the presence of microbial life on the surface or subsurface of Mars. Chemical thermodynamic calculations indicate that on both Earth and Mars, certain gases should exist in extremely low concentrations, if at all. Microbial metabolic activity is an important non-equilibrium chemistry process on Earth, and if microbial life exists on Mars, may be an important nonequilibrium chemistry process on Mars. The non-equilibrium chemistry of the atmosphere of Mars is discussed in this paper.

J S Levine↗

Detecting Amino Acids on Mars

Understanding the events that led to the origin of life on Earth is complicated by the lack of geological evidence from the period around four billion years (4 Gyr) ago when the transition from prebiotic chemistry to biochemistry is believed to be occurred. Although erosion and plate tectonics have since erased the terrestrial geological record from the time of the origin of life, there is possibility that information about this period of Earth history may still be preserved on Mars. A major goal of the NASA Space Exploration Program is to search for evidence of abiotic chemistry and extinct or extant life on Mars. During the next decade, spacecraft will orbit Mars, land on the surface, and return with surface samples for analysis. The question is what compounds should we search for, either directly on the planet or in samples returned to Earth, that will answer unambiguously whether abiotic and/or biotic organic molecules are present.

Bada, Jeffrey L.↗

Detection of microbes in the subsurface

The search for evidence of microbial life in the deep subsurface of Earth has implications for the Mars Rover Sampling Return Missions program. If suitably protected environments can be found on Mars then the instrumentation to detect biomarkers could be used to examine the molecular details. Finding a lipid in Martian soil would represent possibly the simplest test for extant or extinct life. A device that could do a rapid extraction possibly using the supercritical fluid technology under development now with a detection of the carbon content would clearly indicate a sample to be returned.

White, David C.↗

UV-Resistant Non-Spore-Forming Bacteria From Spacecraft-Assembly Facilities

Four species of non-spore-forming bacteria collected from clean-room surfaces in spacecraft-assembly facilities could survive doses of ultraviolet (UV) radiation that would suffice to kill most known cultivable bacterial species. In a previous study, high UV resistance was found in spores of the SAFR-032 strain of Bacillus pumilus, as reported in "Ultraviolet- Resistant Bacterial Spores," NASA Tech Briefs, Vol. 31, No. 9 (September 2007), page 94. These studies are parts of a continuing effort to understand the survival of hardy species of bacteria under harsh conditions, and develop means of sterilizing spacecraft to prevent biocontamination of Mars that could in turn interfere with future life detection missions. The four species investigated were Arthrobacter sp. KSC_Ak2i, Microbacterium schleiferi LMA_AkK1, Brevundimonas diminuta KSC_Ak3a, and Sphingomonas trueperi JSC_Ak7-3. In the study, cells of these species were mixed into Atacama Desert soil (to elucidate the shadowing effect of soil particles) and the resulting mixtures were tested both in solution and in a desiccated state under simulated Martian atmospheric and UV conditions. The UV-survival indices of Arthrobacter sp. and Microbacterium schleiferi were found to be comparable to those of Bacillus pumilus spores.

Venkateswaran, Kasthuri↗

Radioactivity of Sediments at the Atacama Rover Astrobiology Drilling Studies Field Site

Introduction: During the 2018 Atacama Rover Astrobiology Drilling Studies (ARADS) field activities, we conducted a survey of surface and subsurface sediment radioactivity. Our objective was to characterize the radiation environment of various near-surface sampling locations, as well as along the entire profile of a 2.60 m deep pit that had been dug near one of the ARADS rover drill site locations. This pit was used to collect samples that have been well characterized [2] for comparison to samples collected and analyzed by the rover.

ARADS↗

Biotechnology Approaches to Life Detection

The direct detection of organic biomarkers for living or fossil microbes on Mars by an in situ instrument is a worthy goal for future lander missions. Several new and innovative biotechnology approaches are being explored. Firstly we have proposed an instrument based on immunological reactions to specific antibodies to cause activation of fluorescent stains. Antibodies are raised or acquired to a variety of general and specific substances that might be in Mars soil. These antibodies are then combined with various fluorescent stains and applied to micron sized numbered spots on a small (2-3 cm) test plate where they become firmly attached after freeze drying. Using technology that has been developed for gene mining in DNA technology up to 10,000 tests per square inch can now be applied to a test plate. On Mars or the planet/moon of interest, a sample of soil from a trench or drill core is extracted with water and/or an organic solvent and ultrasonication and then applied to the test plate. Any substance, which has an antibody on the test plate, will react with its antibody and activate its fluorescent stain. At the moment a small UV light source will illuminate the test plate, which is observed with a small CCD camera, although other detection systems will be applied. The numbered spots that fluoresce indicate the presence of the tested-for substance, and the intensity indicates relative amounts. Furthermore with up to a thousand test plates available false positives and several variations of antibody can also be screened for. The entire instrument can be quite small and light, on the order of 10 cm in each dimension. A possible choice for light source may be small UV lasers at several wavelengths. Some of the wells or spots can contain simply standard fluorescent stains used to detect live cells, dead cells, DNA, etc. The stains in these spots may be directly activated, with no antibodies being necessary. The proposed system will look for three classes of biomarkers: those from extant life, such as DNA, those from extinct life such as hopanes, and those from organic compounds not necessarily associated with life such as PAHs, rocket exhaust contamination and other a/pre-biotic chemicals. Both monoclonal and polyclonal antibodies can be used. Monoclonal antibodies react with a very specific compound, but polyclonal antibodies may react to any of a whole family of compounds. Furthermore the technique of phage display to raise antibodies against classically non-antigenic molecules is also being considered. Additional information is contained in the original extended abstract.

Steele, Andrew↗

Microbial Habitability in Gale Crater: Sample Analysis at Mars (SAM) Instrument Detection of Microbial Essential Carbon and Nitrogen

Chemical analyses of Mars soils and sediments from previous landed missions have demonstrated that Mars surface materials possessed major (e.g., P, K, Ca, Mg, S) and minor (e.g., Fe, Mn, Zn, Ni, Cl) elements essential to support microbial life. However, the detection of microbial essential organic-carbon (C) and nitrate have been more elusive until the Mars Science Laboratory (MSL) rover mission. Nitrate and organic-C in Gale Crater, Mars have been detected by the Sample Analysis at Mars (SAM) instrument onboard the MSL Curiosity rover. Eolian fines and drilled sedimentary rock samples were heated in the SAM oven from approximately 30 to 860 degrees Centigrade where evolved gases (e.g., nitrous oxide (NO) and CO2) were released and analyzed by SAM’s quadrupole mass spectrometer (MS). The temperatures of evolved NO was assigned to nitrate while evolved CO2 was assigned to organic-C and carbonate. The CO2 releases in several samples occurred below 450 degrees Centigrade suggesting organic-C dominated in those samples. As much as 7 micromoles NO3-N per gram and 200 micromoles CO2-C per gram have been detected in the Gale Crater materials. These N and C levels coupled with assumed microbial biomass (9 x 10 (sup -7) micrograms per cell) C (0.5 micrograms C per micrograms cell) and N (0.14 micrograms N per micrograms cell) requirements, suggests that less than 1 percent and less than 10 percent of Gale Crater C and N, respectively, would be required if available, to accommodate biomass requirements of 1 by 10 (sup 5) cells per gram sediment. While nitrogen is the limiting nutrient, the potential exists that sufficient N and organic-C were present to support limited heterotrophic microbial populations that may have existed on ancient Mars.

Sutter, B.↗

The MASSE Project: Applications of Biotechnology for Planetary Exploration

Automated life-detection experiments for solar system exploration have been previously. proposed and used onboard the. Viking, Mars lander,s, although. with ambiguous results. The recent advances in biotechnology such as biosensors, protein microarrays, and microfluidics alongside increased. knowledge in biomarker science have led to vastly improved sophistication and sensitivity for a new approach in life detection. The MASSE project has taken the challenge of integrating all of this knowledge into a new generation of interplanetary flight instrumentation for the main purpose.ot combining several mutually. confirming tests for life, organic/microbial contamination, prebiotic and abiotic chemicals into a small low powered instrument. Although the primary goal is interplanetary exploration, several terrestrial applications have become apparent specifically in point-of-care medical technology, bio-warfare, environmental sensing and microbial monitoring of manned space-flight vehicles.

Lynch, Kennda↗

Mars biological laboratory.

Biological laboratory development for unmanned exploration of Mars, considering ecology sensing and detection of past or present life

Tompkins, D. N.↗

A Sterile Robotic Mars Soil Analyzer

Since the 1976 Viking Mission to Mars, follow-on efforts to resolve its controversial life detection results have been thwarted by two heretofore insurmountable difficulties: the huge expense of sterilizing the entire spacecraft to protect the integrity of life detection experiments; and the lack of a practical robotic life detection package that could produce results acceptable as unambiguous by the scientific community. We here present a method that assures sterility and the complete integrity of robotic life detection experiments, all at a negligible cost. Second, we propose a candidate set of integrated, highly sensitive experiments that we believe could produce results acceptable to the vast majority of scientists. In addition to the biology-chemistry issue, the extensively debated oxidative state of the Martian surface and other chemical and physical characteristics of the Martian soil would be determined. We present our concept for a miniaturized instrument that could carry out a number of candidate experiments to achieve the objective.

Gilbert V Levin↗

Lanthanide Complexes as a Test for Evidence of Life

The objective of this research is to advance the understanding of the interaction of lanthanide metals with biological organic molecules and to develop a technique to detect these compounds in the solid state and in situ in Mars and other planetary bodies. The detection of these complexes should provide evidence of life past or present. In addition, detection of the metals alone will provide important information about the geological history of a planetary body. Lanthanides were chosen as our focus of interest because they form very stable complexes with organic molecules in solution and they produce intense luminescence in the ultraviolet and visible spectra. The rare earth complexes available are mostly synthetic for diverse applications in medicine. There is not much work done on the complexes that form in nature. Lanthanides have many applications and they are mined aR over the world, however, since the interest has been only in the elements, the analytical techniques employed destroy any organic ligands that may be present. In order to determine if and which lanthanide complexes form in nature and their concentration, soil samples have been collected from areas rich in soluble lanthanide compounds like phosphates and also rich in vegetation. The soil samples will be analyzed and the lanthanide complexes if present will be isolated and characterized. A spectrometer to detect the lanthanide complexes in situ and in the solid state will be designed. In this workshop, the research approach and its implications will be discussed.

Benavides, Jeannette↗

A preliminary study of a Mars penetrator system for subsurface exobiological exploration

The earth seems to stand alone among the planets of the solar system in that its surface, atmosphere, and hydrosphere provide an environment conducive to the maintenance of life. At one time, Mars was also considered to have the potential to harbor life. However, the absence of detectable organic molecules at the two Viking landing sites and the absence of liquid water anywhere on the surface seems to indicate that life is not present on Mars now. There are, however, many indications that the surface of Mars was less hostile in the distant past and the possibility that life may have existed on primordial Mars or that prebiotic chemistry may have occurred can not be excluded. The search for the organic evidence of this prebiotic or biotic activity will require a subsurface study of possible sediments at several sites. The purpose of this paper is to propose and describe a device termed a penetrator to emplant the required instrumentation beneath the surface (2-4 m) at likely sites to search for the evidence of biogenic organic material. The paper examines, in detail, the rationale for the scientific activity, any issues of technical feasibility, and describes the associated system requirements.

Swenson, Byron L.↗

Maximum Sample Temperature for Mars Sample Return: A Historical Perspective

Since the first Mars Sample Return (MSR) report published by the Jet Propulsion Laboratory (JPL) in 1974 [1], a series of panels, reports, and white papers have recognized the importance of sample temperature and offered an informed sample maximum temperature (henceforth SMT) limit for returning martian samples to Earth. The Mars Sample Handling and Requirements Panel (MSHARP, 1999) stated that "[t]he main issue in sample preservation is temperature" [2]. More recently, the Mars Exploration Program Analysis Group (MEPAG)'s "Science Priorities for Mars Sample Return" report (2008), declared that "[s]ignificant loss, particularly to biological studies, occurs if samples reach +50C for three hours", whereby "scientific objectives related to life goals could be seriously compromised" [3]. By contrast, the Mars 2020 mission has adopted a SMT of +60C as spelled out in Beaty et al., 2016 [4]. Samples will be collected and then deposited on the surface in sealed tubes for possible retrieval and return to Earth. Beaty et al. [4] calculates that the samples will experience maximum temperatures of ~+30 to +60C, depending on latitude. At present, there is no mission requirement for the measurement/data logging of sample temperature during this period. We will explore the history of martian SMTs, as they have been recorded since 1974 [1], effectively representing input across multiple generations of Mars scientists. Ten separate publications present SMTs for MSR samples [1-10]. One report [10] is for a mission concept specifically designed to exclude life detection investigations, and recommended an SMT of 50C. Another did not specify a temperature, recommending "Mars ambient temperature" [5]. Of the remaining eight, SMTs are given as: -30C [1], -20C [3], 60C [4], -73 to 41C depending on sample type [6], -40C [7], -43 to 13C depending on type [2,8], and -33C [9]. If we restrict the temperatures to samples highlighted in the Mars 2020 mission goals, i.e. organics-bearing and sedimentary rocks, then the average SMT is -28+/-39C (n=8). Applying a Dixon's Q Test at P=0.05 (two-tailed), the 60C SMT [4] fails with Q=0.602 versus Qcrit=0.526. Excluding the outlier produces an average SMT of -40+/-17C (n=7). Therefore, the average SMT expressed by the Mars science community over the past 44 years (two generations) is a sample temperature no greater than -40C. The difference in chemical reaction rates between this average SMT and Beaty et al [4] can be estimated using the Arrhenius equation. Assuming a generic chemical reaction with an activation energy of 50 kJ/mol and a pre-exponential factor invariant with temperature, this reaction will proceed 2300x faster at 60C than at -40C. To illustrate the effects of the increased reaction rate, consider 10 ppb of alanine in a Mars 2020 cache, and assume that it becomes unmeasurable if it degrades to 1 ppb, as per the Mars 2020 Organic Contamination Panel contamination limits [11]. If we illustrate the effect with an arbitrary degradation rate such that the alanine will become undetectable in ten years at -40C, then the same 10 ppb alanine degrades beyond detectability in only 38 days at 60C. Further research is required to quantify expected analyte losses in the cached samples due to thermal processing.

Fries, Marc↗

A search for life on earth at 100 meter resolution.

Study of several thousand photos indicating that about 0.01 of Gemini and Apollo photographs of the earth at 100 m resolution reveal signs of life - rectangular arrays due to human agricultural and urban territoriality, roads, canals, jet contrails, and industrial pollution. Potential false positives - e.g., dunes, sand bars, jetstream clouds - abound. A curve is derived for the detectivity of contemporary life on earth, in a plot of ground resolution versus global coverage. A comparable biology on Mars would not have been detected by all observations of Mars through Mariner 7.

Sagan, C.↗

Identifying the Effects of X-Ray Computed Tomography on Mars 2020 Tier I Organic Compounds

Mars sample return presents unique challenges for the clean collection, containment, curation and processing of samples. The related issues of life detection and Planetary Protection are of particular importance when developing successful strategies for the acquisition and handling of Mars returned samples. In order to achieve the Mars Sample Return (MSR) science goals, reliable analyses will depend on overcoming some challenging signal/noise-related issues, such that sparse Martian organic compounds will need to be reliably analyzed against the contamination background arising from the complicated MSR campaign. Reliable analyses will depend on clean acquisition, as well as robust documentation of all aspects of both the development and management of the cache.

Weizenbach, L. C.↗

Detection of metabolically produced labeled gas - The Viking Mars Lander.

A qualitative, nonspecific method will test for life on Mars in 1976 by supplying radioactive substrates to samples of the planetary surface material. If microorganisms are present, they may assimilate one or more of the simple labeled compounds and produce radioactive gas. The compounds have been selected on the basis of biological theory and terrestrial results. The measurement of radioactive gas evolved as a function of time constitutes evidence for life. A control performed on a duplicate, but heat sterilized, sample will confirm the biological nature of the results. The shape of the response curve obtained from the viable sample may provide information on the physiological state and generation period of the organisms. Data obtained from a wide variety of terrestrial soils demonstrate a rapid response and high sensitivity for the experiment. Its ability to make comparative studies of soil microorganisms is also demonstrated.

Levin, G. V.↗

Biological Contamination of Mars: Issues and Recommendations

The ad hoc Task Group on Planetary Protection formed by the Space Studies Board (SSB) of the National Research Council focused on making recommendations concerning the protection of Mars from forward contamination (i.e., Earth to Mars) during upcoming missions by both the United States and the former Soviet Union. In so doing, it distinguished between missions whose goals include reconnaissance and measurement and those that specifically include experiments to detect life. The task group also discussed what additional knowledge will be needed in order to assure that future recommendations regarding contamination of Earth from Mars might be made with a higher degree of certainty than is now possible. Following a short introduction to the rationale underlying planetary exploration (Chapter 1) is a brief summary of approved and contemplated missions to Mars (Chapter 2). Chapter 3 briefly reviews the state of knowledge in several areas pertinent to the problem of planetary protection, in the limits of life on Earth and the abilities of known terrestrial organisms to withstand extreme environment conditions, as well as new approaches to detecting life forms. Chapter 5 includes a review and comments (made in light of current knowledge)- on the recommendations made in 'Recommendations on Quarantine Policy for Mars, Jupiter, Saturn, Uranus, Neptune, and Titan'. Updates to the recommendations made in 1978 are also given in Chapter 5. Chapter 6 gives additional recommendations concerning collection of essential data, spacecraft sterilization and bioburden assessment, and future research, as well as legal and social issues and NASA's overall planetary protection program.

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