Chamber for Simulating Martian and Terrestrial Environments

An apparatus for simulating the environment at the surface of Mars has been developed. Within the apparatus, the pressure, gas composition, and temperature of the atmosphere; the incident solar visible and ultraviolet (UV) light; and the attenuation of the light by dust in the atmosphere can be simulated accurately for any latitude, season, or obliquity cycle over the entire geological history of Mars. The apparatus also incorporates instrumentation for monitoring chemical reactions in the simulated atmosphere. The apparatus can be used for experiments in astrobiology, geochemistry, aerobiology, and aerochemistry related to envisioned robotic and human exploration of Mars. Moreover, the apparatus can be easily adapted to enable similar experimentation under environmental conditions of (1) the surfaces of moons, asteroids, and comets, and (2) the upper atmospheres of planets other than Mars: in particular, it can be made to simulate conditions anywhere in the terrestrial atmosphere at altitudes up to about 100 km

Twenty-three Species of Hypobarophilic Bacteria Recovered from Diverse Ecosystems Exhibit Growth under Simulated Martian Conditions at 0.7 kPa

Bacterial growth at low pressure is a new research area with implications for predicting microbial activity in clouds, the bulk atmosphere on Earth, and for modeling the forward contamination of planetary surfaces like Mars. Here we describe experiments on the recovery and identification of 23 species of bacterial hypobarophiles (def., growth under hypobaric conditions of approximately 1-2 kPa) in 11 genera capable of growth at 0.7 kPa. Hypobarophilic bacteria, but not archaea or fungi, were recovered from soil and non-soil ecosystems. The highest numbers of hypobarophiles were recovered from Arctic soil, Siberian permafrost, and human saliva. Isolates were identified through 16S rRNA sequencing to belong to the genera Carnobacterium, Exiguobacterium, Leuconostoc, Paenibacillus, and Trichococcus. The highest population of culturable hypobarophilic bacteria (5.1 x 104 cfu/g) was recovered from Colour Lake soils from Axel Heiberg Island in the Canadian arctic. In addition, we extend the number of hypobarophilic species in the genus Serratia to 6 type-strains that include S. ficaria, S. fonticola, S. grimesii, S. liquefaciens, S. plymuthica, and S. quinivorans. Microbial growth at 0.7 kPa suggests that pressure alone will not be growth-limiting on the martian surface, or in Earth's atmosphere up to an altitude of 34 km

Survivability of Psychrobacter cryohalolentis K5 Under Simulated Martian Surface Conditions

Spacecraft launched to Mars can retain viable terrestrial microorganisms on board that may survive the interplanetary transit. Such biota might compromise the search for life beyond Earth if capable of propagating on Mars. The current study explored the survivability of Psychrobacter cryohalolentis K5, a psychrotolerant microorganism obtained from a Siberian permafrost cryopeg, under simulated martian surface conditions of high ultraviolet irradiation, high desiccation, low temperature, and low atmospheric pressure. First, a desiccation experiment compared the survival of P. cryohalolentis cells embedded, or not embedded, within a medium/salt matrix (MSM) maintained at 25 degrees C for 24 hr within a laminar flow hood. Results indicate that the presence of the MSM enhanced survival of the bacterial cells by 1 to 3 orders of magnitude. Second, tests were conducted in a Mars Simulation Chamber to determine the UV tolerance of the microorganism. No viable vegetative cells of P. cryohalolentis were detected after 8 hr of exposure to Mars-normal conditions of 4.55 W/m(2) UVC irradiation (200-280 nm), -12.5 degrees C, 7.1 mbar, and a Mars gas mix composed of CO2 (95.3%), N2 (2.7%), Ar (1.6%), O2 (0.2%), and H(2)O (0.03%). Third, an experiment was conducted within the Mars chamber in which total atmospheric opacities were simulated at tau = 0.1 (dust-free CO2 atmosphere at 7.1 mbar), 0.5 (normal clear sky with 0.4 = dust opacity and 0.1 = CO2-only opacity), and 3.5 (global dust storm) to determine the survivability of P. cryohalolentis to partially shielded UVC radiation. The survivability of the bacterium increased with the level of UVC attenuation, though population levels still declined several orders of magnitude compared to UVC-absent controls over an 8 hr exposure period

EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements-and to protect our science from human contamination-we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. Engineers at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each attachment. This allowed the tool handle to snap onto a fresh swab attachment much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab attachment is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and repressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between various pressure environments. To further minimize cost, the design team acquired extensive ground test experience in a relevant flight environment by piggy-backing onto suited crew training runs. These training runs allowed the project to validate tool interfaces with pressurized EVA gloves and collect user feedback on the tool design and function, as well as characterize baseline microbial data for different types of spacesuits. In general, test subjects found the EVA Swab Kit relatively straightforward to operate, but identified a number of design improvements that will be incorporated into the final design. Although originally intended to help characterize human forward contaminants, this tool has other potential applications, such as for collecting and preserving space-exposed materials to support astrobiology experiments

Methane from UV-irradiated carbonaceous chondrites under simulated Martian conditions

A UV photolytic process was studied for the production of methane from carbonaceous chondrites under simulated Martian conditions. Methane evolution rates from carbonaceous chondrites were found to be positively correlated to temperature (−80 to 20°C) and the concentration of carbon in the chondrites (0.2 to 1.69 wt%); and decreased over time with Murchison samples exposed to Martian conditions. The amount of evolved methane (EM) per unit of UV energy was 7.9 × 10−13 mol J−1 for UV irradiation of Murchison (1.69 wt%) samples tested under Martian conditions (6.9 mbar and 20°C). Using a previously described Mars UV model (Moores et al., 2007), and the EM given above, an annual interplanetary dust particle (IDP) accreted mass of 2.4 × 105 kg carbon per year yields methane abundances between 2.2 to 11 ppbv for model scenarios in which 20 to 100% of the accreted carbon is converted to methane, respectively. The UV/CH4 model for accreted IDPs can explain a portion of the globally averaged methane abundance on Mars, but cannot easily explain seasonal, temporal, diurnal, or plume fluctuations of methane. Several impact processes were modeled to determine if periodic emplacement of organics from carbonaceous bolides could be invoked to explain the occurrence of methane plumes produced by the UV/CH4process. Modeling of surface impacts of high-density bolides, single airbursts of low-density bolides, and multiple airbursts of a cascading breakup of a low-density rubble-pile comet were all unable to reproduce a methane plume of 45 ppbv, as reported by Mumma et al

Effects of a simulated martian UV flux on the cyanobacterium Chroococcidiopsis 029

ABSTRACT Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4,6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars

The zero forcing numbers and propagation times of gear graphs and helm graphs

Zero forcing is a dynamic coloring process on graphs. Initially, each vertex of a graph is assigned a color of either blue or white, and then a process begins by which blue vertices force white vertices to become blue. The zero forcing number is the cardinality of the smallest set of initially blue vertices which can force the entire graph to become blue, and the propagation time is the minimum number of steps in such a zero forcing process. In this paper we will determine the zero forcing numbers and propagation times of two infinite classes of graphs called gear graphs and helm graphs

Survival of endospores of Bacillus subtilis on spacecraft surfaces under simulated Martian environments: implications for the forward contamination of Mars

Abstract Experiments were conducted in a Mars simulation chamber (MSC) to characterize the survival of endospores of Bacillus subtilis under high UV irradiation and simulated martian conditions. The MSC was used to create Mars surface environments in which pressure (8.5 mb), temperature (−80, −40, −10, or +23 • C), gas composition (Earth-normal N 2 /O 2 mix, pure N 2 , pure CO 2 , or a Mars gas mix), and UV-VIS-NIR fluence rates (200-1200 nm) were maintained within tight limits. The Mars gas mix was composed of CO 2 (95.3%), N 2 (2.7%), Ar (1.7%), O 2 (0.2%), and water vapor (0.03%). Experiments were conducted to measure the effects of pressure, gas composition, and temperature alone or in combination with Mars-normal UV-VIS-NIR light environments. Endospores of B. subtilis, were deposited on aluminum coupons as monolayers in which the average density applied to coupons was 2.47 × 10 6 bacteria per sample. Populations of B. subtilis placed on aluminum coupons and subjected to an Earth-normal temperature (23 • C), pressure (1013 mb), and gas mix (normal N 2 /O 2 ratio) but illuminated with a Mars-normal UV-VIS-NIR spectrum were reduced by over 99.9% after 30 sec exposure to Mars-normal UV fluence rates. However, it required at least 15 min of Mars-normal UV exposure to reduce bacterial populations on aluminum coupons to non-recoverable levels. These results were duplicated when bacteria were exposed to Mars-normal environments of temperature (−10 • C), pressure (8.5 mb), gas composition (pure CO 2 ), and UV fluence rates. In other experiments, results indicated that the gas composition of the atmosphere and the temperature of the bacterial monolayers at the time of Mars UV exposure had no effects on the survival of bacterial endospores. But Mars-normal pressures (8.5 mb) were found to reduce survival by approximately 20-35% compared to Earth-normal pressures (1013 mb). The primary implications of these results are (a) that greater than 99.9% of bacterial populations on sun-exposed surfaces of spacecraft are likely to be inactivated within a few tens of seconds to a few minutes on the surface of Mars, and (b) that within a single Mars day under clear-sky conditions bacterial populations on sun-exposed surfaces of spacecraft will be sterilized. Furthermore, these results suggest that the high UV fluence rates on the martian surface can be an important resource in minimizing the forward contamination of Mars

Bacillus subtilis Spore Resistance to Simulated Mars Surface Conditions

In a Mars exploration scenario, knowing if and how highly resistant Bacillus subtilis spores would survive on the Martian surface is crucial to design planetary protection measures and avoid false positives in life-detection experiments. Therefore, in this study a systematic screening was performed to determine whether B. subtilis spores could survive an average day on Mars. For that, spores from two comprehensive sets of isogenic B. subtilis mutant strains, defective in DNA protection or repair genes, were exposed to 24 h of simulated Martian atmospheric environment with or without 8 h of Martian UV radiation [M(+)UV and M(-)UV, respectively]. When exposed to M(+)UV, spore survival was dependent on: (1) core dehydration maintenance, (2) protection of DNA by α/β-type small acid soluble proteins (SASP), and (3) removal and repair of the major UV photoproduct (SP) in spore DNA. In turn, when exposed to M(-)UV, spore survival was mainly dependent on protection by the multilayered spore coat, and DNA double-strand breaks represent the main lesion accumulated. Bacillus subtilis spores were able to survive for at least a limited time in a simulated Martian environment, both with or without solar UV radiation. Moreover, M(-)UV-treated spores exhibited survival rates significantly higher than the M(+)UV-treated spores. This suggests that on a real Martian surface, radiation shielding of spores (e.g., by dust, rocks, or spacecraft surface irregularities) might significantly extend survival rates. Mutagenesis were strongly dependent on the functionality of all structural components with small acid-soluble spore proteins, coat layers and dipicolinic acid as key protectants and efficiency DNA damage removal by AP endonucleases (ExoA and Nfo), non-homologous end joining (NHEJ), mismatch repair (MMR) and error-prone translesion synthesis (TLS). Thus, future efforts should focus on: (1) determining the DNA damage in wild-type spores exposed to M(+/-)UV and (2) assessing spore survival and viability with shielding of spores via Mars regolith and other relevant materials