Mars surface conditions where liquid water is absent were simulated for the purposes of laboratory research. A pressure-temperature (P-T) profile was maintained in which no combination of pressure or temperature corresponds to the liquid region of the water phase diagram. The triple point of pure water occurs at T = 0.1oC and P(H2O) = 6.01 mbar; therefore all temperatures and pressures must be kept below these values, respectively. A 35-day test was performed in a commercial planetary simulation system (Techshot, Inc., Greenville, IN) in which the minimum night-time temperature was -80oC, the maximum daytime temperature was +26oC, the simulated day-night light cycle in earth hours was 12-on and 12-off, and the total pressure of the pure CO2 atmosphere was maintained below 11 mbar. Any water present was allowed to equilibrate with the changing temperature and pressure. The gas phase was sampled into a CR1-A condensation-mirror low-pressure hygrometer, which uses liquid nitrogen (down to 77oK) to determine the dew point (Buck Technologies, Boulder, CO). Dew point was measured once every hour and recorded on a data logger, along with the varying temperature in the chamber, from which the partial pressure of water was calculated. The resulting calculated daily cycles were tracked on the water P-T diagram, and no points were found to fall within the liquid-phase region of the diagram. It is concluded that there was no liquid water present throughout the test except during the initial pump-down period when aqueous specimens were introduced on the first day (less than 1 hour). Mars regolith simulant was present during this test, and further investigation is needed to determine whether liquid water could have been present or absent in the regolith in the form of brine. Biological samples consisting of Cyanobacteria: Anabena sp., Chroococcidiopsis CCMEE171, Plectonema boryanum; Eubacteria: Bacillus subtilis, Pseudomonas aeruginosa, and Eukaryota: Chlorella ellipsoidia were maintained in the simulator under the above-described conditions. The exposed specimens were tested for intracellular esterase activity, chlorophyll content (where appropriate) and reproductive survival. All tests yielded low-level positive results in all cases. In parallel to these terrestrial studies a planned design study was undertaken for the proposed test bed. Design requirements include compact assembly for transport and installation on the planetary surface (multiple units per mission would be expected), protective internal package for the release of organisms, a means of atmosphere exchange, access to sunlight, a means of penetrating the planetary surface, and most importantly a means of acquiring regolith while meeting the requirements of planetary protection. In consultation with advisers a design was created, and a large-scale mock-up of this design was fabricated by additive manufacturing at Techshot, Inc. with moving parts that simulated the components of the design. The mock-up assembly has been demonstrated to interested parties. A means of detecting live metabolism will also be included in the test bed. Several options were reviewed, and it is concluded that, by the time the ecopoiesis test bed is ready for testing the optimum instrument will be the equivalent of a hand-held mass spectrometer for metabolic gas analysis. This will maximize versatility and reveal much more information than could a detector of a single product (such as molecular oxygen), and the
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