In Situ Real-Time Quantification of Microbial Communities: Applications to Cold and Dry Volcanic Habitats

We report field tests of an instrument using multi-wavelength excitation and detection of fluorescence capable of detection and discrimination of viable cells, non-viable cells (not metabolically active but not decomposed), and spores in extreme arid environments where low microbial abundances are expected. These new results are presented for cold and dry volcanic habitats worldwide, e.g., the arid core of the Atacama Desert, Mt. Kilimanjaro glacier and Kibo area, Pali Aike caldera, and the western US volcanic and desert soils in Utah, Idaho, Nevada, and California. Our results are comparable to previous studies reported in the literature for the same environments. We find these extreme environments there have a base level of ∼103 - 104 cells/g. This is the lower limit of detectable life on terrestrial soils, as we did not observe any quantities less than this, even though the described instrumentation is capable of such measurements. Samples from more conventional environments show much higher microbial cell densities, ca. 108 cells/g or higher, with this same instrument. This base level of microbial life is nearly equal in all the measurements from the extreme environments both hot and cold, and is likely controlled primarily by the sparse nutrients rather than temperature

In Situ Real-Time Quantification of Microbial Communities: Applications to Cold and Dry Volcanic Habitats

We report field tests of an instrument using multi-wavelength excitation and detection of fluorescence capable of detection and discrimination of viable cells, non-viable cells (not metabolically active but not decomposed), and spores in extreme arid environments where low microbial abundances are expected. These new results are presented for cold and dry volcanic habitats worldwide, e.g., the arid core of the Atacama Desert, Mt. Kilimanjaro glacier and Kibo area, Pali Aike caldera, and the western US volcanic and desert soils in Utah, Idaho, Nevada, and California. Our results are comparable to previous studies reported in the literature for the same environments. We find these extreme environments there have a base level of ∼103 - 104 cells/g. This is the lower limit of detectable life on terrestrial soils, as we did not observe any quantities less than this, even though the described instrumentation is capable of such measurements. Samples from more conventional environments show much higher microbial cell densities, ca. 108 cells/g or higher, with this same instrument. This base level of microbial life is nearly equal in all the measurements from the extreme environments both hot and cold, and is likely controlled primarily by the sparse nutrients rather than temperature

The Sample Handling System for the Mars Icebreaker Life Mission: from Dirt to Data

The Mars icebreaker life mission will search for subsurface life on mars. It consists of three payload elements: a drill to retrieve soil samples from approx. 1 meter below the surface, a robotic sample handling system to deliver the sample from the drill to the instruments, and the instruments themselves. This paper will discuss the robotic sample handling system

Ladakh: Diverse, high-altitude extreme environments for off-earth analogue and astrobiology research

This paper highlights unique sites in Ladakh, India, investigated during our 2016 multidisciplinary pathfinding expedition to the region. We summarize our scientific findings and the site's potential to support science exploration, testing of new technologies and science protocols within the framework of astrobiology research. Ladakh has several accessible, diverse, pristine and extreme environments at very high altitudes (3000-5700 m above sea level). These sites include glacial passes, sand dunes, hot springs and saline lake shorelines with periglacial features. We report geological observations and environmental characteristics (of astrobiological significance) along with the development of regolith-landform maps for cold high passes. The effects of the diurnal water cycle on salt deliquescence were studied using the ExoMars Mission instrument mockup: HabitAbility: Brines, Irradiance and Temperature (HABIT). It recorded the existence of an interaction between the diurnal water cycle in the atmosphere and salts in the soil (which can serve as habitable liquid water reservoirs). Life detection assays were also tested to establish the best protocols for biomass measurements in brines, periglacial ice-mud and permafrost melt water environments in the Tso-Kar region. This campaign helped confirm the relevance of clays and brines as interest targets of research on Mars for biomarker preservation and life detection.The team would like to express its gratitude to BirbalSahni Institute of Palaeosciences, Department of Science and Technology,Office of Chief Wildlife Warden of Ladakh, Government of India for helpingarrange the requisite clearances and permits for the conducted work. Projectmentoring and guidance provided by Spaceward Bound members at NASAAmes Research Center. Financial and logistics support provided by TataMotors Ltd, Inspired Journeys Co, Pearl Travels Ltd and NationalGeographic Traveller India. Website and IT support provided by the BlueMarble Space Institute of Science. Audio-video documentation support pro-vided by Astroproject India and The H

SOLID2: An Antibody Array-Based Life-Detector Instrument in a Mars Drilling Simulation Experiment (MARTE)

A field prototype of an antibody array-based life-detector instrument, Signs Of LIfe Detector (SOLID2), has been tested in a Mars drilling mission simulation called MARTE (Mars Astrobiology Research and Technology Experiment). As one of the analytical instruments on the MARTE robotic drilling rig, SOLID2 performed automatic sample processing and analysis of ground core samples (0.5 g) with protein microarrays that contained 157 different antibodies. Core samples from different depths (down to 5.5 m) were analyzed, and positive reactions were obtained in antibodies raised against the Gram-negative bacterium Leptospirillum ferrooxidans, a species of the genus Acidithiobacillus (both common microorganisms in the Río Tinto area), and extracts from biofilms and other natural samples from the Río Tinto area. These positive reactions were absent when the samples were previously subjected to a high-temperature treatment, which indicates the biological origin and structural dependency of the antibody-antigen reactions. We conclude that an antibody array-based life-detector instrument like SOLID2 can detect complex biological material, and it should be considered as a potential analytical instrument for future planetary missions that search for life.The work was funded by the Spanish Ministerio de Educación y Ciencia (ESP2004-05008), the Centro de Astrobiología (CAB), the Instituto Nacional de Técnica Aeroespacial (INTA), and by the National Aeronautics and Space Administration (NASA) Astrobiology Science and Technology for Exploring Planets (ASTEP) program under NRA 02-OSS-01.Peer reviewe

Subsurface scientific exploration of extraterrestrial environments (MINAR 5): analogue science, technology and education in the Boulby Mine, UK

The deep subsurface of other planetary bodies is of special interest for robotic and human exploration. The subsurface provides access to planetary interior processes, thus yielding insights into planetary formation and evolution. On Mars, the subsurface might harbour the most habitable conditions. In the context of human exploration, the subsurface can provide refugia for habitation from extreme surface conditions. We describe the fifth Mine Analogue Research (MINAR 5) programme at 1 km depth in the Boulby Mine, UK in collaboration with Spaceward Bound NASA and the Kalam Centre, India, to test instruments and methods for the robotic and human exploration of deep environments on the Moon and Mars. The geological context in Permian evaporites provides an analogue to evaporitic materials on other planetary bodies such as Mars. A wide range of sample acquisition instruments (NASA drills, Small Planetary Impulse Tool (SPLIT) robotic hammer, universal sampling bags), analytical instruments (Raman spectroscopy, Close-Up Imager, Minion DNA sequencing technology, methane stable isotope analysis, biomolecule and metabolic life detection instruments) and environmental monitoring equipment (passive air particle sampler, particle detectors and environmental monitoring equipment) was deployed in an integrated campaign. Investigations included studying the geochemical signatures of chloride and sulphate evaporitic minerals, testing methods for life detection and planetary protection around human-tended operations, and investigations on the radiation environment of the deep subsurface. The MINAR analogue activity occurs in an active mine, showing how the development of space exploration technology can be used to contribute to addressing immediate Earth-based challenges. During the campaign, in collaboration with European Space Agency (ESA), MINAR was used for astronaut familiarization with future exploration tools and techniques. The campaign was used to develop primary and secondary school and primary to secondary transition curriculum materials on-site during the campaign which was focused on a classroom extra vehicular activity simulation

In Situ Real-Time Monitoring for Aseptic Drilling: Lessons Learned from the Atacama Rover Astrobiology Drilling Studies Contamination Control Strategy and Implementation and Application to the Icebreaker Mars Life Detection Mission

In 2019, the Atacama Rover Astrobiology Drilling Studies (ARADS) project field-tested an autonomous rover-mounted robotic drill prototype for a 6-Sol life detection mission to Mars (Icebreaker). ARADS drilled Mars-like materials in the Atacama Desert (Chile), one of the most life-diminished regions on Earth, where mitigating contamination transfer into life-detection instruments becomes critical. Our Contamination Control Strategy and Implementation (CCSI) for the Sample Handling and Transfer System (SHTS) hardware (drill, scoop and funnels) included out-of-simulation protocol testing (out-of-sim) for hardware decontamination and verification during the 6-Sol simulation (in-sim). The most effective five-step decontamination combined safer-to-use sterilants (3%_hydrogen-peroxide-activated 5%_sodium-hypochlorite), and in situ real-time verification by adenosine triphosphate (ATP) and Signs of Life Detector (SOLID) Fluorescence Immunoassay characterization of hardware bioburden and airborne contaminants. The 20- to 40-min protocol enabled a 4-log bioburden reduction down to <0.1 fmoles ATP detection limit (funnels and drill) to 0.2–0.7 fmoles (scoop) of total ATP. The (post-cleaning) hardware background was 0.3 to 1–2 attomoles ATP/cm2 (cleanliness benchmark background values) equivalent to ca. 1–10 colony forming unit (CFU)/cm2. Further, 60–100% of the in-sim hardware background was ≤3–4 bacterial cells/cm2, the threshold limit for Class <7 aseptic operations. Across the six Sols, the flux of airborne contaminants to the drill sites was ∼5 and ∼22 amoles ATP/(cm2·day), accounting for an unexpectedly high Fluorescence Intensity (FI) signal (FI: ∼6000) against aquatic cyanobacteria, but negligible anthropogenic contribution. The SOLID immunoassay also detected microorganisms from multiple habitats across the Atacama Desert (anoxic, alkaline/acidic microenvironments in halite fields, playas, and alluvial fans) in both airborne and post-cleaning hardware background. Finally, the hardware ATP background was 40–250 times lower than the ATP in cores. Similarly, the FI peaks (FImax) against the microbial taxa and molecular biomarkers detected in the post-cleaned hardware (FI: ∼1500–1600) were 5–10 times lower than biomarkers in drilled sediments, excluding significant interference with putative biomarker found in cores. Similar protocols enable the acquisition of contamination-free materials for ultra-sensitive instruments analysis and the integrity of scientific results. Their application can augment our scientific knowledge of the distribution of cryptic life on Mars-like grounds and support life-detection robotic and human-operated missions to Mars.This work was funded in part by the National Aeronautics and Space Administration PSTAR program project number 14-PSTAR142-0032 to Brian Glass. ARADS was a project under the auspices of the NASA PSTAR (NNH14ZDA001N-PSTAR) Program led by Dr. Mary Voytek. Support of the SOLID/LDChip instruments came from Grant number RTI2018-094368-B-I00 (SOLID) and MDM-2017-0737 under the Unidad de Excelencia “Maria de Maeztu” Centro de Astrobiología (CSIC-INTA) program by the Spanish Ministry of Science and Innovation/State Agency of Research (MCIN/AEI/5 10.13039/501100011033) and also with support from “ERDF: A way of making Europe.”Peer reviewe

MARTE: Technology development and lessons learned from a mars drilling mission simulation

29 pages, 21 figures, 2 tables.-- ISI Article Identifier: 000250768000006.-- Special issue: Mining Robotics.The NASA Mars Astrobiology Research and Technology Experiment (MARTE) performed a field test simulating a robotic drilling mission on Mars in September 2005. The experiment took place in Minas de Riotinto in southwestern Spain, a highly relevant Mars analog site. The experiment utilized a 10 m class dry auger coring drill, a robotic core sample handling system, onboard science and life detection instruments, and a borehole inspection probe, all of which were mounted to a simulated lander platform. Much of the operation of the system was automated, and the resulting data were transmitted via satellite to remote science teams for analysis. The science team used the data to characterize the subsurface geology and to search for signs of life. Based on the data being received and operational constraints, the science team also directed the daily operation of the equipment. The experiment was highly successful, with the drill reaching over 6 m in depth in 23 days of simulated mission. The science team analyzed remote sensing data obtained from 28 cores and detected biosignatures in 12 core subsamples. This experiment represents an important first step in understanding the technology and operational requirements for a future Mars drilling mission. In the past there have been numerous rover field tests that have helped guide the design and implementation of the highly successful rover missions to Mars. However, a drilling mission potentially adds a new level of complexity, and it is important to understand the associated challenges. This paper documents the design of the experimental system, highlighting some of the more important design criteria and design trades. It also discusses the results of the field testing and lists some of the key technological lessons learned.MARTE was jointly funded by the NASA Astrobiology Science and Technology for Exploring Planets (ASTEP) program through NRA 02-OSS-01. Partial funding for automation was provided by the NASA Intelligent Systems program. Spanish participation in MARTE was funded by the Centro de Astrobiología. We also thank the Museo Minero de Rio Tinto, Spain and the staff of the Vazquez Díaz Hotel in Nerva, Spain for providing field facilities and logistical support to the MARTE project.Peer reviewe