Planetary Protection Knowledge Gap Closure Enabling Crewed Missions to Mars.

As focus for exploration of Mars transitions from current robotic explorers to development of crewed missions, it remains important to protect the integrity of scientific investigations at Mars, as well as protect the Earth's biosphere from any potential harmful effects from returned martian material. This is the discipline of planetary protection, and the Committee on Space Research (COSPAR) maintains the consensus international policy and guidelines on how this is implemented. Based on National Aeronautics and Space Administration (NASA) and European Space Agency (ESA) studies that began in 2001, COSPAR adopted principles and guidelines for human missions to Mars in 2008. At that point, it was clear that to move from those qualitative provisions, a great deal of work and interaction with spacecraft designers would be necessary to generate meaningful quantitative recommendations that could embody the intent of the Outer Space Treaty (Article IX) in the design of such missions. Beginning in 2016, COSPAR then sponsored a multiyear interdisciplinary meeting series to address planetary protection "knowledge gaps" (KGs) with the intent of adapting and extending the current robotic mission-focused Planetary Protection Policy to support the design and implementation of crewed and hybrid exploration missions. This article describes the outcome of the interdisciplinary COSPAR meeting series, to describe and address these KGs, as well as identify potential paths to gap closure. It includes the background scientific basis for each topic area and knowledge updates since the meeting series ended. In particular, credible solutions for KG closure are described for the three topic areas of (1) microbial monitoring of spacecraft and crew health; (2) natural transport (and survival) of terrestrial microbial contamination at Mars, and (3) the technology and operation of spacecraft systems for contamination control. The article includes a KG data table on these topic areas, which is intended to be a point of departure for making future progress in developing an end-to-end planetary protection requirements implementation solution for a crewed mission to Mars. Overall, the workshop series has provided evidence of the feasibility of planetary protection implementation for a crewed Mars mission, given (1) the establishment of needed zoning, emission, transport, and survival parameters for terrestrial biological contamination and (2) the creation of an accepted risk-based compliance approach for adoption by spacefaring actors including national space agencies and commercial/nongovernment organizations.

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 for characterization 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.

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 approximately 1 m 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. Collecting samples from ice-rich soils on Mars in search of life presents two challenges: protection of that icy soil--considered a "special region" with respect to planetary protection--from contamination from Earth, and delivery of the icy, sticky soil to spacecraft instruments. We present a sampling device that meets these challenges. We built a prototype system and tested it at martian pressure, drilling into ice-cemented soil, collecting cuttings, and transferring them to the inlet port of the SOLID2 life-detection instrument. The tests successfully demonstrated that the Icebreaker drill, sample handling system, and life-detection instrument can collectively operate in these conditions and produce science data that can be delivered via telemetry--from dirt to data. Our results also demonstrate the feasibility of using an air gap to prevent forward contamination. We define a set of six analog soils for testing over a range of soil cohesion, from loose sand to basalt soil, with angles of repose of 27° and 39°, respectively. Particle size is a key determinant of jamming of mechanical parts by soil particles. Jamming occurs when the clearance between moving parts is equal in size to the most common particle size or equal to three of these particles together. Three particles acting together tend to form bridges and lead to clogging. Our experiments show that rotary-hammer action of the Icebreaker drill influences the particle size, typically reducing particle size by ≈ 100 µm.

Astrobiology through the ages of Mars: the study of terrestrial analogues to understand the habitability of Mars.

Mars has undergone three main climatic stages throughout its geological history, beginning with a water-rich epoch, followed by a cold and semi-arid era, and transitioning into present-day arid and very cold desert conditions. These global climatic eras also represent three different stages of planetary habitability: an early, potentially habitable stage when the basic requisites for life as we know it were present (liquid water and energy); an intermediate extreme stage, when liquid solutions became scarce or very challenging for life; and the most recent stage during which conditions on the surface have been largely uninhabitable, except perhaps in some isolated niches. Our understanding of the evolution of Mars is now sufficient to assign specific terrestrial environments to each of these periods. Through the study of Mars terrestrial analogues, we have assessed and constrained the habitability conditions for each of these stages, the geochemistry of the surface, and the likelihood for the preservation of organic and inorganic biosignatures. The study of these analog environments provides important information to better understand past and current mission results as well as to support the design and selection of instruments and the planning for future exploratory missions to Mars.

Design and practices for use of automated drilling and sample handling in MARTE while minimizing terrestrial and cross contamination.

Mars Astrobiology Research and Technology Experiment (MARTE) investigators used an automated drill and sample processing hardware to detect and categorize life-forms found in subsurface rock at Río Tinto, Spain. For the science to be successful, it was necessary for the biomass from other sources--whether from previously processed samples (cross contamination) or the terrestrial environment (forward contamination)-to be insignificant. The hardware and practices used in MARTE were designed around this problem. Here, we describe some of the design issues that were faced and classify them into problems that are unique to terrestrial tests versus problems that would also exist for a system that was flown to Mars. Assessment of the biomass at various stages in the sample handling process revealed mixed results; the instrument design seemed to minimize cross contamination, but contamination from the surrounding environment sometimes made its way onto the surface of samples. Techniques used during the MARTE Río Tinto project, such as facing the sample, appear to remove this environmental contamination without introducing significant cross contamination from previous samples.

Science results from a Mars drilling simulation (Río Tinto, Spain) and ground truth for remote science observations.

Science results from a field-simulated lander payload and post-mission laboratory investigations provided "ground truth" to interpret remote science observations made as part of the 2005 Mars Astrobiology Research and Technology Experiment (MARTE) drilling mission simulation. The experiment was successful in detecting evidence for life, habitability, and preservation potential of organics in a relevant astrobiological analogue of Mars. SCIENCE RESULTS: Borehole 7 was drilled near the Río Tinto headwaters at Peña de Hierro (Spain) in the upper oxidized remnant of an acid rock drainage system. Analysis of 29 cores (215 cm of core was recovered from 606 cm penetrated depth) revealed a matrix of goethite- (42-94%) and hematite-rich (47-87%) rocks with pockets of phyllosilicates (47-74%) and fine- to coarse-grained loose material. Post-mission X-ray diffraction (XRD) analysis confirmed the range of hematite:goethite mixtures that were visually recognizable (approximately 1:1, approximately 1:2, and approximately 1:3 mixtures displayed a yellowish-red color whereas 3:1 mixtures displayed a dark reddish-brown color). Organic carbon was poorly preserved in hematite/goethite-rich materials (C(org) <0.05 wt %) beneath the biologically active organic-rich soil horizon (C(org) approximately 3-11 wt %) in contrast to the phyllosilicate-rich zones (C(org) approximately 0.23 wt %). GROUND TRUTH VS. REMOTE SCIENCE ANALYSIS: Laboratory-based analytical results were compared to the analyses obtained by a Remote Science Team (RST) using a blind protocol. Ferric iron phases, lithostratigraphy, and inferred geologic history were correctly identified by the RST with the exception of phyllosilicate-rich materials that were misinterpreted as weathered igneous rock. Adenosine 5'-triphosphate (ATP) luminometry, a tool available to the RST, revealed ATP amounts above background noise, i.e., 278-876 Relative Luminosity Units (RLUs) in only 6 cores, whereas organic carbon was detected in all cores. Our manned vs. remote observations based on automated acquisitions during the project provide insights for the preparation of future astrobiology-driven Mars missions.

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 2005 MARTE Robotic Drilling Experiment in Río Tinto, Spain: objectives, approach, and results of a simulated mission to search for life in the Martian subsurface.

The Mars Astrobiology Research and Technology Experiment (MARTE) simulated a robotic drilling mission to search for subsurface life on Mars. The drill site was on Peña de Hierro near the headwaters of the Río Tinto river (southwest Spain), on a deposit that includes massive sulfides and their gossanized remains that resemble some iron and sulfur minerals found on Mars. The mission used a fluidless, 10-axis, autonomous coring drill mounted on a simulated lander. Cores were faced; then instruments collected color wide-angle context images, color microscopic images, visible-near infrared point spectra, and (lower resolution) visible-near infrared hyperspectral images. Cores were then stored for further processing or ejected. A borehole inspection system collected panoramic imaging and Raman spectra of borehole walls. Life detection was performed on full cores with an adenosine triphosphate luciferin-luciferase bioluminescence assay and on crushed core sections with SOLID2, an antibody array-based instrument. Two remotely located science teams analyzed the remote sensing data and chose subsample locations. In 30 days of operation, the drill penetrated to 6 m and collected 21 cores. Biosignatures were detected in 12 of 15 samples analyzed by SOLID2. Science teams correctly interpreted the nature of the deposits drilled as compared to the ground truth. This experiment shows that drilling to search for subsurface life on Mars is technically feasible and scientifically rewarding.