The National Aeronautics and Space Administration (NASA) and the European Space Agency (ESA) are studying how samples might be brought back to Earth from Mars safely. Backward planetary protection is key in this complex endeavour, as it is required to prevent potential adverse effects from returning materials to Earth's biosphere. As the question of whether or not life exists on Mars today or whether it ever did in the past is still unanswered, the effort to return samples from Mars is expected to be categorized as a ‘Restricted Earth Return’ mission, for which NASA policy requires the containment of any unsterilized material returned to Earth. NASA is investigating several solutions to contain Mars samples and sterilize any uncontained Martian particles. This effort has significant implications for both NASA's scientific mission, and the Earth's environment; and so special care and vigilance are needed in planning and execution in order to assure acceptance of safety to Earth's biosphere. To generate a technically acceptable sterilization process across a wide array of scientific and other stakeholders, on 30–31 January 2019, 10–11 June 2019 and 19–20 February 2020, NASA informally convened a Sterilization Working Group (SWG) composed of experts from industry, academia and government to assess methods for sterilization and inactivation, to identify future work needed to verify these methods against biological challenges, and to determine their feasibility for implementation on robotic spacecraft in deep space. The goals of the SWG were:

1. (1) Understand what it means to sterilize and/or inactivate Martian materials and how that understanding can be applied to the Mars Sample Return (MSR) mission.

2. (2) Assess methods for sterilization and inactivation, and identify future work needed to verify these methods.

3. (3) Provide an effective plan for communicating with other agencies and the public.

This paper provides a summary of the discussions and conclusions of the SWG over these three workshops. It reflects a consensus position based on qualitative discussion of how agencies might approach the problem of sterilization of Mars material. The SWG reached a consensus that sterilization options can be considered on the basis of biology as we know it, and that sterilization modalities that are effective on terrestrial materials and organisms should be part of the MSR planetary protection strategy. Conclusions pointed to several industry standards for sterilization to include heat, chemical, UV radiation and low-heat plasma. Technical trade-offs for each sterilization modality were discussed while simultaneously considering the engineering challenges and limitations for spaceflight. Future work includes more in-depth discussions on technical trade-offs of sterilization modalities, identifying and testing Earth analogue challenge organisms and proteinaceous molecules against chosen modalities, and executing collaborative agreements between NASA and external working group partners to help close data gaps, and to establish strong, scientifically grounded sterilization and inactivation standards for MSR.

The presence of perchlorate in the Martian soil may limit in-situ resource utilization (ISRU) technologies to support human outposts. In order to exploit the desiccation, radiation-tolerant cyanobacterium Chroococcidopsis in Biological Life Support Systems based on ISRU, we investigated the perchlorate tolerance of Chroococcidopsis sp. CCMEE 029 and its derivative CCMEE 029 P-MRS. This strain was obtained from dried cells mixed with Martian regolith simulant and exposed to Mars-like conditions during the BIOMEX space experiment. After a 55-day exposure of up to 200 mM perchlorate ions, a tolerance threshold value of 100 mM perchlorate ions was identified for both Chroococcidopsis strains. After 40-day incubation, a Mars-relevant perchlorate concentration of 2.4 mM perchlorate ions, provided as a 60 and 40% mixture of Mg- and Ca-perchlorate, had no negative effect on the growth rate of the two strains. A proof-of-concept experiment was conducted using Chroococcidopsis lysate in ISRU technologies to feed a heterotrophic bacterium, i.e. an Escherichia coli strain capable of metabolizing sucrose. The sucrose content was fivefold increased in Chroococcidopsis cells through air-drying and the yielded lysate successfully supported the bacterial growth. This suggested that Chroococcidopsis is a suitable candidate for ISRU technologies to support heterotrophic BLSS components in a Mars-relevant perchlorate environment that would prove challenging to many other cyanobacteria, allowing a ‘live off the land’ approach on Mars.

Previously conducted space missions revealed the presence of perchlorates, which are known to have a high oxidizing potential in Martian regolith, at the level of 0.5%. Due to hygroscopic properties and crystallization features of perchlorate-containing solutions, assumptions leading to the possibility of the existence of liquid water in the form of brines, which can contribute to the vital activity of microorganisms, have been made. At the same time, high concentrations of perchlorates can inhibit the growth of microorganisms and cause their death. Previously performed studies have discovered the presence of highly diverse microbial communities in terrestrial perchlorate-containing soils and have also demonstrated the stability and activity of some prokaryotes cultured on highly concentrated perchlorates media (over 10%). Nevertheless, the limits of microbial tolerance to perchlorates and whether microbial communities are able to withstand the effects of high concentrations of perchlorates remain uncertain. The aim of this research was to study the reaction of microbial communities of hot-arid and cryo-arid soils and sedimentary rocks to the adding of a highly concentrated solution of sodium perchlorate (5%) in situ. An increase in the total number of prokaryotes, the number of metabolically active Bacteria and Archaea, and the variety of the consumed substrates were revealed. It was observed that in samples incubated with sodium perchlorate, a high taxonomic diversity of the microbial community is preserved at a level comparable to control sample. The study shows that the presence of high concentrations of sodium perchlorate (5%) in the soil does not lead to the death or significant inhibition of microbial communities.

Planetary habitability may be affected by exposure to γ radiation from supernovae (SNe). Records of Earth history during the late Quaternary Period (40 000 years to present) allow testing for specific SN γ radiation effects. SNe include Type Ia white dwarf explosions, Type Ib, c and II core collapses, and many γ burst objects. Surveys of galactic SNe remnants offer a nearly complete accounting for this time and including SN distances and ages. Terrestrial changes in records of the cosmogenic isotope 14C are here compared to SN-predicted changes. SN γ emission occurs mainly within 3 years; average per-event total emissions of 4 × 1049 erg are used for comparison of close events There are 18 SNe ≤ 1.5 kpc, and brief 14C anomalies are reported for eight of the closest. Four are notable (BP is year before 1950 CE): the older Vela SNR and an abrupt 30‰ del 14C rise at 12 740 BP; S165 and a 20‰ rise at 7431 BP; Vela Jr. and a 14‰ rise at 2765 BP; and HB9 and a 9‰ rise at 5372 BP. Rapid-increase anomalies in 14C production have been attributed to cosmic rays from exceptionally large solar flares. However, the proximity and ages of these SNe, the probable size and duration of their γ emissions, the predicted effects on 14C, and the agreement with 14C records together support SNe causation. Also, the supposed solar-caused 14C anomalies at CE 774 and 993 may instead have been caused by the SNe associated with the G190.9-2.2 and G347.3-00.5 remnants. Both are of appropriate age and distance.

The formation mechanism of linear and isopropyl cyanide (hereafter n-PrCN and i-PrCN, respectively) in the interstellar medium (ISM) has been proposed from the reaction between some previously detected small cyanides/cyanide radicals and hydrocarbons/hydrocarbon radicals. n-PrCN and i-PrCN are nitriles therefore, they can be precursors of amino acids via Strecker synthesis. The chemistry of i-PrCN is especially important since it is the first and only branched molecule in ISM, hence, it could be a precursor of branched amino acids such as leucine, isoleucine, etc. Therefore, both n-PrCN and i-PrCN have significant astrobiological importance. To study the formation of n-PrCN and i-PrCN in ISM, quantum chemical calculations have been performed using density functional theory at the MP2/6-311++G(2d,p)//M062X/6-311+G(2d,p) level. All the proposed reactions have been studied in the gas phase and the interstellar water ice. It is found that reactions of small cyanide with hydrocarbon radicals result in the formation of either large cyanide radicals or ethyl and vinyl cyanide, both of which are very important prebiotic interstellar species. They subsequently react with the radicals CH2 and CH3 to yield n-PrCN and i-PrCN. The proposed reactions are efficient in the hot cores of SgrB2 (N) (where both n-PrCN and i-PrCN were detected) due to either being barrierless or due to the presence of a permeable entrance barrier. However, the formation of n-PrCN and i-PrCN from the ethyl and vinyl cyanide always has an entrance barrier impermeable in the dark cloud; therefore, our proposed pathways are inefficient in the deep regions of molecular clouds. It is also observed that ethyl and vinyl cyanide serve as direct precursors to n-PrCN and i-PrCN and their abundance in ISM is directly related to the abundance of both isomers of propyl cyanide in ISM. In all the cases, reactions in the ice have smaller barriers compared to their gas-phase counterparts.

Approximately 22% of sun-like stars have Earth-like exoplanets. Advanced civilizations may exist on these, and significant effort has been expended on the theoretical analysis of planetary systems, and accompanying practical detection instruments.

The longevity of technological civilizations is unknown, as is the probability of less advanced societies becoming technological. Accordingly, searching for pre-industrial extra-terrestrial societies may be more productive.

Using the earth as a model, a consideration of possible detectible proxies suggests that observation of seasonal agriculture may be possible in the near future – particularly in ideal circumstances, for which quantitative analysis is provided. More speculatively, other detectible processes may include: species introduction; climate change; large urban fires and land-use or aquatic changes.

Primitive societies may be both aware that their activities may be observed from other planets, and may deliberately adjust these activities to aid or conceal detection.

The origin of life from inanimate matter is still an open question, and our knowledge is still very limited. In this sense, prebiotic chemistry seeks to study and understand how chemical reactions may have contributed to the origin of life. Minerals are of great relevance to prebiotic chemistry, as they may have preconcentrated precursors of biomolecules or biomolecules from diluted solutions, provided protection for biomolecules against UV radiation and hydrolysis, catalysing their reactions and played the role of a primitive genetic code. Montmorillonite, a prebiotic mineral, was shown to be able to adsorb adenine and later also histidine. In addition, histidine adsorption did not displace adenine from the montmorillonite. Kinetic experiments showed that using a whole period of time (7 days) it was not possible to adjust the data to any mathematical kinetic model. Thus, the data were separated into four different adsorption ranges: range 1 (0–60 min), range 2 (60–4320 min), range 3 (4320–7200 min) and range 4 (7200–10 080 min). Range 1 showed adsorption that was too fast, meaning no variations in the adsorption data, and the data of range 3 did not fit in any model used in this work. Thus, range 2 (60–4320 min) and range 4 (7200–10 080 min) were analysed. The adsorption kinetics of histidine adsorption indicated two reaction steps, a quick step (60–4320 min), following the pseudo-first-order model, followed by a slower step (7200–10 080 min) of the pseudo-second order. With these results, isotherms were constructed with times of 1 h and 7 days. The results of the quick step (1 h) showed a reaction that was not thermodynamically favoured. For this time range, Gibbs energy values obtained ranged between 5 and 10 kJ mol−1 at temperatures of 20, 35 and 50°C, and the adsorption occurred due to the balance shift of increase in histidine concentrations. The isotherms of the slow step (7 days) presented negative values, showing a more favourable reaction with Gibbs energy values ranging between −5 and −11 kJ mol−1. The mathematical modelling of the data indicates that seawater ions are crucial in the adsorption process. Thus, the study provided essential information for prebiotic chemistry, showing that time and the reaction medium should always be taken into account.

Geomorphological studies of the hidden and protected subsurface environments are crucial to obtain a greater insight into the evolution of planetary landforms, hydrology, climate, geology and mineralogy. From an astrobiological point of view subsurface environments are of interest for their potential habitability as they are local environments that are partially or fully shielded from the high levels of space and solar radiation. Furthermore, in the case of Mars, there is an increasing interest in searching for the presence of past or extant life in its subsurface. These applications make it mandatory to investigate equipment and instrumentation that allow for the study of subsurface geomorphology, as well as organic chemical biomarkers, such as biomolecules, carbon, nitrogen and sulphur isotopes, and other biologically significant minerals and gases. Mines on Earth can be used as analogues to investigate the geomorphology of Martian subsurface environments and perform astrobiology studies. With that goal, we have developed a low-cost, robust, remotely operable subsurface rover called KORE (KOmpact Rover for Exploration). This work illustrates the studies of a terrestrial analogue for the exploration of Mars using KORE during the Mine Analogue Research 6 (MINAR 6) campaign with the low-cost 3D mapping technology InXSpace 3D (In situ 3D mapping tool eXploration of space 3D). InXSpace 3D utilizes an RGB-D camera that captures depth information in addition to the RGB data of an image, operating based on the structured light principle capable of providing depth information in mm scale resolution at sub 3 m mapping range. InXSpace 3D is used to capture point clouds of natural and artificial features, thereby obtaining information about geologically relevant structures and also to incorporate them in earth mining safety. We tested two of the dense simultaneous localization and mapping (SLAM) algorithms: Kintinuous and Real-Time Appearance-Based Mapping (RTAB-Map) to check the performance of InXSpace 3D in a dark mine environment. Also, the air accumulation of volatiles such as methane and formaldehyde due to thermogenic and mining process was measured with the environmental station payload on the rover platform, which caters to both astrobiological significance and mine safety. The main conclusions of this work are: (1) a comparison made between the RTAB-Map algorithm and Kintinuous algorithm showed the superiority of Kintinuous algorithm in providing better 3D reconstruction; although RTAB-Map algorithm captured more points than the Kintinuous algorithm in the dark mine environment; (2) a comparison of point cloud images captured with and without lighting conditions had a negligible effect on the surface density of the point clouds; (3) close-range imaging of the polygonal features occurring on the halite walls using InXSpace 3D provided mm-scale resolution to enable further characterization; (4) heuristic algorithms to quickly post-process the 3D point cloud data provided encouraging results for preliminary analyses; (5) we successfully demonstrated the application of KORE to mine safety; and (6) the multi-sensors platform on KORE successfully monitored the accumulated volatiles in the mine atmosphere during its operation. The findings obtained during this KORE campaign could be incorporated in designing and planning future subsurface rover explorations to potential planetary bodies such as Mars with synergistic applications to subsurface environments in mines on Earth.