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It may be that habitable planets are common but life is rare. If future advances in telescopes increasingly suggest this is so, humankind might feel motivated to seed lifeless planets with resilient terrestrial organisms or synthetic forms designed to thrive on the target planet. A useful mechanism for achieving this goal at a relatively low cost is to use interstellar comets transiting the Solar System to convey microbial cargoes toward nearby planetary systems, where they could disseminate the inoculum via their dust trails. Conversely, it is conceivable that terrestrial life was deliberately seeded in this matter, a hypothesis that could be tested if we found evidence for life on other Solar System bodies that displayed common basic biochemical signatures. Our scenario raises a number of ethical and technological challenges that need to be addressed.

Microbiological activities can be detected in various extreme environments on Earth, which suggest that extraterrestrial environments, such as on Mars, could host life. There have been proposed a number of biomarkers to detect extant life mostly based on specific molecules. Because terrestrial organisms have catalytic proteins (enzymes), enzymatic activity may also be a good indicator to evaluate biological activities in extreme environments. Phosphatases are essential for all terrestrial organisms because phosphate esters are ubiquitously used in genetic molecules (DNA/RNA) and membranes. In this study, we evaluated microbial activity in soils of the Atacama Desert, Chile, by analysing several biomarkers, including phosphatase activity. Phosphatases extracted with Tris buffer were assayed fluorometrically using 4-methylumbelliferyl phosphate as a substrate. The horizontal distribution of phosphatase activity and other parameters in soils from the Atacama Desert showed that phosphatase activity was positively correlated with amino acid concentration and colony-forming units and negatively correlated with precipitation amount. We found consistent that biochemical indicators including phosphatase significantly decreased in the extreme hyper-arid zone where rainfall of −1. The results were compared with phosphatase activities detected in extreme environments, such as submarine hydrothermal systems and Antarctic soils, as well as soils from ordinary environments. Overall, our results suggested that phosphatase activity could be a good indicator for evaluating biological activities in extreme environments.

We report 3 years of data from one meteorological and three smaller stations in University Valley, a high-elevation (1677 m) site in the Dry Valleys of Antarctica with extensive dry permafrost. Mean air temperature was -23.4°C. Summer air temperatures were virtually always < 0°C and were consistent with the altitude lapse rate and empirical relationships between summer temperature, distance from the coast and elevation. The measured frost point (-22.5°C) at the 42 cm deep ice table is equal to the surface frost point and above the atmospheric frost point (-29.6°C), providing direct evidence that surface conditions control ground ice depth. Observed peak surface soil temperatures reach 6°C for ice-cemented ground > 15 cm deep but stay < 0°C when it is shallower. We develop an energy balance model tuned to this rocky and dry environment. We find that differences in peak soil surface temperatures are primarily due to the higher thermal diffusivity of ice-cemented ground compared to dry soil. Sensitivity studies show that expected natural variability is insufficient for melt to form and significant excursions from current conditions are required. The site's ice table meets the criteria for a Special Region on Mars, with 30% of the year > -18°C and water activity > 0.6.

Plant growth experiments on near-term lunar landers need to be relatively small, lightweight, and self-contained. Here, we report on the design of a ~1 liter volume (1U Cubesat size) hermetically sealed habitat suitable for plant growth experiments during the first 10 days of seedling development of Arabidopsis thaliana and Brassica nigra. Images from a single interior camera show germination and provide quantitative data on seedling height, leaf area, and circumnutations. After 10 days with illumination from LEDs, the photosynthetic area of Arabidopsis cotyledons per seedling reached 300 mm2. Seedling height, inferred from the overhead camera using reference markers, reached was 15 ± 5 mm. Robust circumnutation in seedlings was observed. CO2 increased as expected due to respiration in the seeds during germination reaching levels of 5000 ppm after 3 days before declining to 3000 ppm on day 10 due to photosynthetic uptake. No CO2 was added to the sealed chamber during the experiments. These results show that fundamental studies of germination and initial growth can be conducted in a small volume (1 L) hermetically sealed unit with only an overhead camera and CO2 sensor. Hardware based on this approach would be suitable for lunar experiments on robotic landers.

The novel extremophilic yeast Rhodotorula frigidialcoholis, formerly R. JG1b, was isolated from ice-cemented permafrost in University Valley (Antarctic), one of coldest and driest environments on Earth. Phenotypic and phylogenetic analyses classified R. frigidialcoholis as a novel species. To characterize its cold-adaptive strategies, we performed mRNA and sRNA transcriptomic analyses, phenotypic profiling, and assessed ethanol production at 0 and 23 °C. Downregulation of the ETC and citrate cycle genes, overexpression of fermentation and pentose phosphate pathways genes, growth without reduction of tetrazolium dye, and our discovery of ethanol production at 0 °C indicate that R. frigidialcoholis induces a metabolic switch from respiration to ethanol fermentation as adaptation in Antarctic permafrost. This is the first report of microbial ethanol fermentation utilized as the major energy pathway in response to cold and the coldest temperature reported for natural ethanol production. R. frigidialcoholis increased its diversity and abundance of sRNAs when grown at 0 versus 23 °C. This was consistent with increase in transcription of Dicer, a key protein for sRNA processing. Our results strongly imply that post-transcriptional regulation of gene expression and mRNA silencing may be a novel evolutionary fungal adaptation in the cryosphere.

Life detection on Mars is an important topic that includes a direct search for biomarkers. This requires instruments for

We report infrared reflectance and ultraviolet fluorescence spectra of the surfaces and cleaved side of Beacon Sandstone from Antarctica that harbours a cryptoendolithic microbial community - a photosynthesis-based consortium of algae, lichen and bacteria present a few millimetres below the surface. Chlorophyll absorptions were present in the reflectance spectra of the exposed interior but not on the top or bottom surfaces and their band depths changed < 4% between measurements taken 19 years apart, indicating the stability of the microorganisms when the sample is kept dry. The presence of subsurface organic layers was detected in reflectance at 3.41 μm on the sample's surface. Fluorescence spectra of the cleaved side showed the blue fluorescence peaks expected from chlorophyll but no 0.65–0.80 μm peaks seen in fluorescence measurements of green vegetation. A weak fluorescence signal was detectable at the surface of the sample, presumably due to some light leaking into the subsurface through pores or cracks in the goethite coating the sample's surface. Theoretically, this weak fluorescence signal could possibly be observed in rock surfaces broken by erosion or meteor impacts on Mars. Sandstone outcrops have been reported on Mars and detection of organic layers in sandstones there would be of interest.

The martian surface has been continuously exposed to galactic cosmic radiation. Since organic compounds are degraded by ionizing radiation, knowledge of their decay constants is fundamental to pred...

Artículo escrito por un elevado número de autores, solo se referencian el que aparece en primer lugar, los autores pertenecientes a la UAM y el nombre del grupo de colaboración, si lo hubiere, The dispersion of microorganisms through the atmosphere is a continual and essential process that underpins biogeography and ecosystem development and function. Despite the ubiquity of atmospheric microorganisms globally, specific knowledge of the determinants of atmospheric microbial diversity at any given location remains unresolved. Here we describe bacterial diversity in the atmospheric boundary layer and underlying soil at twelve globally distributed locations encompassing all major biomes, and characterise the contribution of local and distant soils to the observed atmospheric community. Across biomes the diversity of bacteria in the atmosphere was negatively correlated with mean annual precipitation but positively correlated to mean annual temperature. We identified distinct non-randomly assembled atmosphere and soil communities from each location, and some broad trends persisted across biomes including the enrichment of desiccation and UV tolerant taxa in the atmospheric community. Source tracking revealed that local soils were more influential than distant soil sources in determining observed diversity in the atmosphere, with more emissive semi-arid and arid biomes contributing most to signatures from distant soil. Our findings highlight complexities in the atmospheric microbiota that are relevant to understanding regional and global ecosystem connectivity, This work was supported by the Singapore Ministry of Education and Yale-NUS College, grant number R-607-265-331-121

There has been considerable attention on how to detect life on other worlds by searching for biomolecules. However, there has been much less clarity as to when it becomes warranted to focus a mission on the search for life on another world. At a minimum, a life-detection mission should follow convincing evidence of (1) Liquid water of suitable salinity, past or present; (2) Carbon in the water; (3) Biologically available N in the water; (4) Biologically useful energy in the water; (5) Organic material that can possibly be of biological origin and a plausible strategy for sampling this material. Based on these prerequisites, the most promising targets for a life search are currently the plume of Enceladus and the subsurface of Mars-in equatorial lake bed sediments and in polar ice-cemented ground. Neither the surface of Europa nor the clouds of Venus meet the criteria listed here but may with further exploration.

The emerging understanding of microbial trace gas chemotrophy as a metabolic strategy to support energy and carbon acquisition for microbial survival and growth has significant implications in the search for past, and even extant, life beyond Earth. The use of trace gases, including hydrogen and carbon monoxide as substrates for microbial oxidation, potentially offers a viable strategy with which to support life on planetary bodies that possess a suitable atmospheric composition, such as Mars and Titan. Here, we discuss the current state of knowledge of this process and explore its potential in the field of astrobiological exploration.

Atmospheric transport is critical to dispersal of microorganisms between habitats and this underpins resilience in terrestrial and marine ecosystems globally. A key unresolved question is whether microorganisms assemble to form a taxonomically distinct, geographically variable, and functionally adapted atmospheric microbiota. Here we characterised inter-continental patterns of microbial taxonomic and functional diversity in air within and above the atmospheric boundary layer and in underlying soils for 596 globally sourced samples. Bacterial and fungal assemblages in air were taxonomically structured and deviated significantly from purely stochastic assembly. Patterns differed with location and reflected underlying surface cover and environmental filtering. Source-tracking indicated a complex recruitment process involving local soils plus globally distributed inputs from drylands and the phyllosphere. Assemblages displayed stress-response and metabolic traits relevant to survival in air, and taxonomic and functional diversity were correlated with macroclimate and atmospheric variables. Our findings highlight complexity in the atmospheric microbiota that is key to understanding regional and global ecosystem connectivity.

A key science priority for planetary exploration is to search for signs of life in our Solar System. Life-detection mission concepts aim to assess whether or not biomolecular signatures of life are present, which requires highly sensitive instrumentation. This introduces greater risk of false positives, and perhaps false negatives. Stringent science-derived contamination requirements for achieving science measurements on life-detection missions necessitate mitigation approaches that minimize, protect from, and prevent science-relevant contamination of critical surfaces of the science payload and provide high confidence to life-detection determinations. To this end, we report on technology advances that focus on understanding contamination transfer from pre-launch processing to end of mission using high-fidelity physics in the form of computational fluid dynamics and sorption physics for monolayer adsorption/desorption, and on developing a new full-spacecraft bio-molecular barrier design that restricts contamination of the spacecraft and instruments by the launch vehicle hardware. The bio-molecular barrier isolates the spacecraft from biological, molecular, and particulate contamination from the external environment. Models were used to evaluate contamination transport for a designs reference mission that utilizes the barrier. Results of the modeling verify the efficacy of the barrier and an in-cruise decontamination activity. Overall mission contamination tracking from launch to science operations demonstrated exceptionally low probability on contamination impacting science measurements, meeting the stringent contamination requirements of femtomolar levels of compounds. These advances will enable planetary missions that aim to detect and identify signatures of life in our Solar System.

Habitability has been generally defined as the capability of an environment to support life. Ecologists have been using Habitat Suitability Models (HSMs) for more than four decades to study the habitability of Earth from local to global scales. Astrobiologists have been proposing different habitability models for some time, with little integration and consistency among them, being different in function to those used by ecologists. Habitability models are not only used to determine if environments are habitable or not, but they also are used to characterize what key factors are responsible for the gradual transition from low to high habitability states. Here we review and compare some of the different models used by ecologists and astrobiologists and suggest how they could be integrated into new habitability standards. Such standards will help to improve the comparison and characterization of potentially habitable environments, prioritize target selections, and study correlations between habitability and biosignatures. Habitability models are the foundation of planetary habitability science and the synergy between ecologists and astrobiologists is necessary to expand our understanding of the habitability of Earth, the Solar System, and extrasolar planets., Published in Astrobiology, 21(8). arXiv admin note: substantial text overlap with arXiv:2007.05491

We have developed a compact instrument called the “COmpact COlor BIofinder”, or CoCoBi, for the standoff detection of biological materials and organics with polyaromatic hydrocarbons (PAHs) using a nondestructive approach in a wide area. The CoCoBi system uses a compact solid state, conductively cooled neodymium-doped yttrium aluminum garnet (Nd:YAG) nanosecond pulsed laser capable of simultaneously providing two excitation wavelengths, 355 and 532 nm, and a compact, sensitive-gated color complementary metal–oxide–semiconductor camera detector. The system is compact, portable, and determines the location of biological materials and organics with PAHs in an area 1590 cm2 wide, from a target distance of 3 m through live video using fast fluorescence signals. The CoCoBi system is highly sensitive and capable of detecting a PAH concentration below 1 part per billion from a distance of 1 m. The color images provide the simultaneous detection of various objects in the target area using shades of color and morphological features. We demonstrate that this unique feature successfully detected the biological remains present in a 150-million-year-old fossil buried in a fluorescent clay matrix. The CoCoBi was also successfully field-tested in Hawaiian ocean water during daylight hours for the detection of natural biological materials present in the ocean. The wide-area and video-speed imaging capabilities of CoCoBi for biodetection may be highly useful in future NASA rover–lander life detection missions.

NASA’s Dragonfly mission will send a rotorcraft lander to the surface of Titan in the mid-2030s. Dragonfly's science themes include investigation of Titan’s prebiotic chemistry, habitability, and potential chemical biosignatures from both water-based “life as we know it” (as might occur in the interior mantle ocean, potential cryovolcanic flows, and/or impact melt deposits) and potential “life, but not as we know it” that might use liquid hydrocarbons as a solvent (within Titan’s lakes, seas, and/or aquifers). Consideration of both of these solvents simultaneously led to our initial landing site in Titan’s equatorial dunes and interdunes to sample organic sediments and water ice, respectively. Ultimately, Dragonfly's traverse target is the 80 km diameter Selk Crater, at 7° N, where we seek previously liquid water that has mixed with surface organics. Our science goals include determining how far prebiotic chemistry has progressed on Titan and what molecules and elements might be available for such chemistry. We will also determine the role of Titan’s tropical deserts in the global methane cycle. We will investigate the processes and processing rates that modify Titan’s surface geology and constrain how and where organics and liquid water can mix on and within Titan. Importantly, we will search for chemical biosignatures indicative of past or extant biological processes. As such, Dragonfly, along with Perseverance, is the first NASA mission to explicitly incorporate the search for signs of life into its mission goals since the Viking landers in 1976.

The recent suggestion of phosphine in Venus’s atmosphere has regenerated interest in the idea of life in clouds. However, such analyses usually neglect the role of water activity, which is a measure of the relative availability of water, in habitability. Here we compute the water activity within the clouds of Venus and other Solar System planets from observations of temperature and water-vapour abundance. We find water-activity values of sulfuric acid droplets, which constitute the bulk of Venus’s clouds, of ≤0.004, two orders of magnitude below the 0.585 limit for known extremophiles. Considering other planets, ice formation on Mars imposes a water activity of ≤0.537, slightly below the habitable range, whereas conditions are biologically permissive (>0.585) at Jupiter’s clouds (although other factors such as their composition may play a role in limiting their habitability). By way of comparison, Earth’s troposphere conditions are, in general, biologically permissive, whereas the atmosphere becomes too dry for active life above the middle stratosphere. The approach used in the current study can also be applied to extrasolar planets., We are grateful to S. L. Clegg (University of East Anglia, England, UK) for helpful discussions on the use of the E-AIM at low water activity and the provision of some code; C. S. Cockell (University of Edinburgh, Scotland, UK), D. Y. Sorokin (Winogradsky Institute of Microbiology, Russia) and A. Ventosa (University of Seville, Spain) for providing information about thermotolerance of halophiles; M. S. Marley (NASA Ames Research Center, CA, USA) for information on Jupiter and exoplanets; A. Méndez (University of Puerto Rico, Puerto Rico) for inputs relating to analysis of Earth’s atmosphere; J. R. Lobry (University of Lyons, France) who helped with use of the cardinal pH model; N. J. Tosca (University of Cambridge, England, UK) for discussions about thermodynamic properties of aqueous sulfuric acid solutions; and E. L. J. Watkin (Curtin University, Australia) who provided information about stress tolerance of Acidihalobacter. J.E.H. was funded by the Biotechnology and Biological Sciences Research Council (BBSRC, United Kingdom) project BBF003471; M.-P.Z. was supported by projects PID2019-104205GB-C21 of Ministry of Science and Innovation and MDM-2017-0737 Unidad de Excelencia ‘María de Maeztu’- Centro de Astrobiología (CSIC-INTA) (Spain); and O.V.G. was supported by the Centre of Environmental Biotechnology Project (grant 810280) funded by the European Regional Development Fund (ERDF) through the Welsh Government.

We summarize the observations and context for molecular hydrogen (H2) in Titan's atmosphere where it is the third most abundant gas. Hydrogen escapes to space but is replenished by methane photochemistry. An open question is whether there are sources and/or sinks in the surface and subsurface: sources might include serpentinization reactions in the deep interior while sinks might involve reactions with acetylene mediated by chemical or even biological catalysts. Cassini data provide weak evidence of a surface sink, and also point to variations with latitude of the tropospheric hydrogen abundance, so further measurements would be of value. We demonstrate that a simple solid-state sensor can provide the required measurement precision in an oxygen-free atmosphere, and consider how measurements on a mobile platform may inform the question of sources and sinks. We underscore the importance of simultaneous methane and hydrogen measurements: whereas the stratosphere is a hydrogen source and methane sink, serpentinization could be a subsurface source of both gasses.

Dry permafrost - ground with temperature always below 0°C and containing negligible ice - overlying ice-cemented ground has been reported in the Dry Valleys of Antarctica and on Mars. Here we report on a new site (79°49.213'S, 83°18.860'W, 718 m elevation) located on the side of Mount Dolence in Ellsworth Land, Antarctica. Year-round temperature and humidity measurements indicate that dry permafrost is present between depths of 13.5 and 49.0 cm - the location of ice-cemented ground. The mean annual frost point of the ice-cemented ground is -17.0 ± 0.2°C and the mean annual frost point of the atmosphere is -22.7 ± 1°C. The corresponding mean annual temperatures are -19.2°C and -20.3°C. Neither the temperature of the ice-cemented ground nor the air rise above freezing. Both the dry permafrost and the ice table may be habitable. In the dry soil at 3 cm depth there are 80 hours in the summer when temperature exceeds -5°C and water activity exceeds 0.8. At the ice table, temperature exceeds -10°C and water activity exceeds 0.8 for 35 hours in the year. The ice table and the dry permafrost above it would be considered a ‘Special Region’ on Mars. Further microbial investigation of this site is indicated.

Lake Untersee is one of the largest perennially ice-covered lakes in Dronning Maud Land. We investigated the energy and water mass balance of Lake Untersee to understand its state of equilibrium. The thickness of the ice cover is strongly correlated with sublimation rates; variations in sublimation rates across the ice cover are largely determined by wind-driven turbulent heat fluxes and the number of snow-covered days. Lake extent and water level have remained stable for the past 20 years, indicating that the water mass balance is in equilibrium. The lake is damned by the Anuchin Glacier and mass balance calculation suggest that subaqueous melting of terminus ice contributes 40–45% of the annual water budget; since there is no evidence of streams flowing into the lake, the lake must be connected to a groundwater system that contributes 55–60% in order to maintain the lake budget in balance. The groundwater likely flows at a rate of ~8.8 × 10−2m3s−1, a reasonable estimate given the range of subglacial water flux in the region. The fate of its well-sealed ice cover is likely tied to changes in wind regime, whereas changes in water budget are more closely linked to the response of surrounding glaciers to climate change.

Humans want to go to Mars now. This entails many difficult but solvable challenges. However, human activity on Mars depends on the answers to three fundamental questions: (1) Is there life ...

Water tracks in the Antarctic Dry Valleys are dark linear features of increased soil moisture that flow downslope over the spring and summer, providing a source of moisture in a cold-arid desert. They are typically sourced from melting snow, ground ice, and deliquescence (Levy et al., 2011). This research presents the first in-depth study of the activity potential and diversity of microbial communities of Antarctic water tracks. We investigated whether these water track soils are more habitable to microbial communities by ascertaining the differences in diversity, total and culturable cell counts, and microbial respiratory activity in water track soils compared with the adjacent dry soils in Pearse Valley. Total cell counts ranged from 1.47 × 10³ to 4.17 × 10⁵ cells/g dry weight soil. Water track soils had higher total and culturable biomass, in addition to higher microbial activity at 5° and −5°C, compared with adjacent dry soils. Microbial respiration was positively correlated with soil moisture content, but total cell counts and plate counts were not. Surprisingly, microbial community composition did not differ between wet and dry soil communities, and was not related to soil moisture content. The microbial community composition instead appeared to differ spatially based on location and depth. Overall, the data suggest that cold water tracks are more habitable than the surrounding cold-arid soils. Our results suggest that recurring slope lineae, which are dark linear features that grow downslope on Mars over the spring and summer, where liquid water might be a recurring phenomenon, could be sites of astrobiological potential.

Shallow Radar soundings of the south polar layered deposits (SPLD) have revealed buried CO2 ice with as many as three distinct layers separated by thinner layers of water ice (Bierson et al., 2016). This layering is suggestive of formation by cyclical processes such as obliquity, eccentricity and advance of perihelion. If obliquity is the main driver, then there were many opportunities for CO2 deposits to have formed within the last 3.5-4 Myr. To persist, however, CO2 deposits must be followed by the deposition of an insulating layer of porous water ice that can seal in the CO2 to survive the obliquity maxima that follow. We suggest that the existing deposits were formed within the last 350 kyr. Each obliquity minimum was quickly followed by a period in which perihelion occurred in the northern summer (southern winter) solstice. Under these conditions, an enhanced deposition of low porosity, fine-grained water ice could occur on the winter pole. A similar series of depositions and burials by water ice could have occurred between 2.75 and 2.2 million years ago, but these deposits were unlikely to have survived the many high obliquity swings that occurred between 2.2 Ma and 400 ka. The formation and burial of CO2 ice could also have occurred in similar fashion earlier during the last ∼ 100 Myr in which the southern polar layered deposits were formed. Although at current times the geothermal flux may be too low to cause basal melting of CO2 deposits, in earlier times, deposits formed in deeper scarps and valleys could have led to basal CO2 melting and sequestration into the regolith. By extension, basal melting and sequestration in the regolith may provide an explanation for the fate of the thick CO2 atmosphere (pCO2 ≳ 0.5 bar) implied by climate models of early Mars.

Lake Untersee is a perennially ice-covered Antarctic lake that consists of two basins. The deepest basin, next to the Anuchin Glacier is aerobic to its maximum depth of 160 m. The shallower basin has a maximum depth of 100 m, is anoxic below 80 m, and is shielded from convective currents. The thermal profile in the anoxic basin is unusual in that the water temperature below 50 m is constant at 4°C but rises to 5°C between 70 m and 80 m depth, then drops to 3.7°C at the bottom. Field measurements were used to conduct a thermal and stability analysis of the anoxic basin. The shape of the thermal maximum implies two discrete locations of energy input, one of 0.11 W m-2 at 71 m depth and one of 0.06 W m-2 at 80 m depth. Heat from microbial activity cannot account for the required amount of energy at either depth. Instead, absorption of solar radiation due to an increase in water opacity at these depths can account for the required energy input. Hence, while microbial metabolism is not an important source of heat, biomass increases opacity in the water column resulting in greater absorption of sunlight.

Preferential uptake of either levorotatory (L) or dextrorotatory (D) enantiomer of a chiral molecule is a potential planetary life-detection method. On Earth, bacteria, as a rule, metabolize D-sugars and L-amino acids. Here, we use growth experiments to identify exceptions to the rule and their potential impact on the method's reliability. Our experiments involve six strains of Bacillus and collective uptake of the sugars glucose and arabinose, and the amino acids alanine, glutamic acid, leucine, cysteine, and serine-all of which are highly soluble. We find that selective uptake is not evident unless (1) each sugar is tested individually and (2) multiple amino acids are tested together in a mixture. Combining sugars should be avoided because, as we show in Bacillus bacteria, the same organisms may catabolize one sugar, glucose, in D-form and another sugar, arabinose, in L-form. Single amino acids should be avoided because bacteria can access certain proteinogenically incompatible enantiomers using specific racemases. Specifically, bacteria contain an alanine acid racemase and can catabolize D-alanine if no other D-amino acids are present. The proposed improvements would reliably separate nonselective chemical reactions from biological reactions and, if life is indicated, inform whether the selective patterns for amino acids and sugars are the same as on Earth.

Enceladus’s long-lived plume of ice grains and water vapor makes accessing oceanic material readily achievable from orbit (around Saturn or Enceladus) and from the moon’s surface. In preparation for the National Academies of Sciences, Engineering and Medicine 2023–2032 Planetary Science and Astrobiology Decadal Survey, we investigated four architectures capable of collecting and analyzing plume material from orbit and/or on the surface to address the most pressing questions at Enceladus: Is the subsurface ocean inhabited? Why, or why not? Trades specific to these four architectures were studied to allow an evaluation of the science return with respect to investment. The team found that Orbilander, a mission concept that would first orbit and then land on Enceladus, represented the best balance. Orbilander was thus studied at a higher fidelity, including a more detailed science operations plan during both orbital and landed phases, landing site characterization and selection analyses, and landing procedures. The Orbilander mission concept demonstrates that scientifically compelling but resource-conscious Flagship-class missions can be executed in the next decade to search for life at Enceladus.