Biological production of H2 , CH4 and CO2 in the deep subsurface of the Iberian Pyrite Belt.

Most of the terrestrial deep subsurfaces are oligotrophic environments in which some gases, mainly H2 , CH4 and CO2 , play an important role as energy and/or carbon sources. In this work, we assessed their biotic and abiotic origin in samples from subsurface hard-rock cores of the Iberian Pyrite Belt (IPB) at three different depths (414, 497 and 520 m). One set of samples was sterilized (abiotic control) and all samples were incubated under anaerobic conditions. Our results showed that H2 , CH4 and CO2 remained low and constant in the sterilized controls while their levels were 4, 4.1 and 2.5 times higher respectively, in the unsterilized samples compared to the abiotic controls. The δ13 CCH4 -values measured in the samples (range -31.2 to -43.0 ‰) reveals carbon isotopic signatures that are within the range for biological methane production. Possible microorganisms responsible for the biotic production of the gases were assessed by CARD-FISH. The analysis of sequenced genomes of detected microorganisms within the subsurface of the IPB allowed to identify possible metabolic activities involved in H2 (Rhodoplanes, Shewanella and Desulfosporosinus), CH4 (Methanobacteriales) and CO2 production. The obtained results suggest that part of the H2 , CH4 and CO2 detected in the deep subsurface has a biological origin.

Transitory microbial habitat in the hyperarid Atacama Desert.

Traces of life are nearly ubiquitous on Earth. However, a central unresolved question is whether these traces always indicate an active microbial community or whether, in extreme environments, such as hyperarid deserts, they instead reflect just dormant or dead cells. Although microbial biomass and diversity decrease with increasing aridity in the Atacama Desert, we provide multiple lines of evidence for the presence of an at times metabolically active, microbial community in one of the driest places on Earth. We base this observation on four major lines of evidence: (i) a physico-chemical characterization of the soil habitability after an exceptional rain event, (ii) identified biomolecules indicative of potentially active cells [e.g., presence of ATP, phospholipid fatty acids (PLFAs), metabolites, and enzymatic activity], (iii) measurements of in situ replication rates of genomes of uncultivated bacteria reconstructed from selected samples, and (iv) microbial community patterns specific to soil parameters and depths. We infer that the microbial populations have undergone selection and adaptation in response to their specific soil microenvironment and in particular to the degree of aridity. Collectively, our results highlight that even the hyperarid Atacama Desert can provide a habitable environment for microorganisms that allows them to become metabolically active following an episodic increase in moisture and that once it decreases, so does the activity of the microbiota. These results have implications for the prospect of life on other planets such as Mars, which has transitioned from an earlier wetter environment to today's extreme hyperaridity.

Interlaboratory test for stable carbon isotope analysis of dissolved inorganic carbon in geothermal fluids.

RATIONALE: Stable carbon isotope ratios have many applications in natural sciences. In the first worldwide interlaboratory proficiency test, the discrepancies in measured δ13 CDIC values of natural waters were up to σ = ±3‰. Therefore, we continued the investigation on the analytical data quality assurance of individual laboratories and internal consistency among laboratories worldwide. METHODS: We designed and performed an interlaboratory comparison exercise for δ13 C analyses of ten water and two solid samples (Na2 CO3 , CaCO3 ), including two synthetic samples prepared by dissolving the carbonates individually. Three laboratories analyzed an additional sample set to assess solution stability, at least one month after the first set analysis period. The δ13 C values were measured using dual inlet isotope ratio mass spectrometry (DI-IRMS) or continuous flow (CF)-IRMS. RESULTS: The δ13 C values of solid Na2 CO3 and its aqueous solution were -5.06 ± 0.21‰ and 5.32 ± 0.24‰, respectively, while the δ13 C value of solid CaCO3 was -4.49 ± 0.93‰. Similarly, the lake water has a consistent value (2.45 ± 0.19‰). The δ13 C values of geothermal water have a wide dispersion among individual laboratory measurements and among those of different laboratories; however, a trend exists in the δ13 C values measured at the three sampling points of each well. CONCLUSIONS: The δ13 C values of solid Na2 CO3 and its solution, and lake water (i.e. DIC concentration samples >100 mg/L carbon) are consistent among all the participating laboratories. The dispersion in the δ13 C values of solid CaCO3 is associated with its lower chemical affinity than that of Na2 CO3 . The poor reproducibility in the δ13 C values of geothermal fluids, collected at three points of a geothermal well, despite overall consistent trends regarding their collection points suggests inadequate sample handling (atmospheric CO2 exchange) and/or inappropriate analytical approaches (incomplete H3 PO4 acid reaction).

A neuropeptide modulates sensory perception in the entomopathogenic nematode Steinernema carpocapsae.

Entomopathogenic nematodes (EPNs) employ a sophisticated chemosensory apparatus to detect potential hosts. Understanding the molecular basis of relevant host-finding behaviours could facilitate improved EPN biocontrol approaches, and could lend insight to similar behaviours in economically important mammalian parasites. FMRFamide-like peptides are enriched and conserved across the Phylum Nematoda, and have been linked with motor and sensory function, including dispersal and aggregating behaviours in the free living nematode Caenorhabditis elegans. The RNA interference (RNAi) pathway of Steinernema carpocapsae was characterised in silico, and employed to knockdown the expression of the FMRFamide-like peptide 21 (GLGPRPLRFamide) gene (flp-21) in S. carpocapsae infective juveniles; a first instance of RNAi in this genus, and a first in an infective juvenile of any EPN species. Our data show that 5 mg/ml dsRNA and 50 mM serotonin triggers statistically significant flp-21 knockdown (-84%***) over a 48 h timecourse, which inhibits host-finding (chemosensory), dispersal, hyperactive nictation and jumping behaviours. However, whilst 1 mg/ml dsRNA and 50 mM serotonin also triggers statistically significant flp-21 knockdown (-51%**) over a 48 h timecourse, it does not trigger the null sensory phenotypes; statistically significant target knockdown can still lead to false negative results, necessitating appropriate experimental design. SPME GC-MS volatile profiles of two EPN hosts, Galleria mellonella and Tenebrio molitor reveal an array of shared and unique compounds; these differences had no impact on null flp-21 RNAi phenotypes for the behaviours assayed. Localisation of flp-21 / FLP-21 to paired anterior neurons by whole mount in situ hybridisation and immunocytochemistry corroborates the RNAi data, further suggesting a role in sensory modulation. These data can underpin efforts to study these behaviours in other economically important parasites, and could facilitate molecular approaches to EPN strain improvement for biocontrol.

Inter-laboratory test for oxygen and hydrogen stable isotope analyses of geothermal fluids: Assessment of reservoir fluid compositions.

RATIONALE: Knowledge of the accuracy and precision for oxygen (δ18 O values) and hydrogen (δ2 H values) stable isotope analyses of geothermal fluid samples is important to understand geothermal reservoir processes, such as partial boiling-condensation and encroachment of cold and reinjected waters. The challenging aspects of the analytical techniques for this specific matrix include memory effects and higher scatter of delta values with increasing total dissolved solids (TDS) concentrations, deterioration of Pt-catalysts by dissolved/gaseous H2 S for hydrogen isotope equilibration measurements and isotope salt effects that offset isotope ratios determined by gas equilibration techniques. METHODS: An inter-laboratory comparison exercise for the determination of the δ18 O and δ2 H values of nine geothermal fluid samples was conducted among eleven laboratories from eight countries (CeMIEGeo2017). The delta values were measured by dual inlet isotope ratio mass spectrometry (DI-IRMS), continuous flow IRMS (CF-IRMS) and/or laser absorption spectroscopy (LAS). Moreover, five of these laboratories analyzed an additional sample set at least one month after the analysis period of the first set. Statistical evaluation of all the results was performed to obtain the expected isotope ratios of each sample, which were then subsequently used in deep reservoir fluid composition calculations. RESULTS: The overall analytical precisions of the measurements were ± 0.2‰ for δ18 O values and ± 2.0‰ for δ2 H values within the 95% confidence interval. CONCLUSIONS: The measured and calculated δ18 O and δ2 H values of water sampled at the weir box, separator and wellhead of geothermal wells suggest the existence of hydrogen and oxygen isotope-exchange equilibrium between the liquid and vapor phases at all sampling points in the well. Thus, both procedures for calculating the isotopic compositions of the deep geothermal reservoir fluid - using either the analytical data of the liquid phase at the weir box together with those of vapor at the separator or the analytical data of liquid and vapor phases at the separator -are equally valid.

Spatial Distributions of DDTs in the Water Masses of the Arctic Ocean.

There is a scarcity of data on the amount and distribution of the organochlorine pesticide dichlorodiphenyltrichloroethane (DDT) and its metabolites in intermediate and deep ocean water masses. Here, the distribution and inventories of DDTs in water of the Arctic shelf seas and the interior basin are presented. The occurrence of ∑6DDT (0.10-66 pg L-1) in the surface water was dominated by 4,4'-DDE. In the Central Arctic Ocean increasing concentrations of DDE with depth were observed in the Makarov and Amundsen basins. The increasing concentrations down to 2500 m depth is in accordance with previous findings for PCBs and PBDEs. Similar concentrations of DDT and DDEs were found in the surface water, while the relative contribution of DDEs increased with depth, demonstrating a transformation over time and depth. Higher concentrations of DDTs were found in the European part of the Arctic Ocean; these distributions likely reflect a combination of different usage patterns, transport, and fate of these compounds. For instance, the elevated concentrations of DDTs in the Barents and Atlantic sectors of the Arctic Ocean indicate the northbound Atlantic current as a significant conveyor of DDTs. This study contributes to the very rare data on OCPs in the vast deep-water compartments and combined with surface water distribution across the Arctic Ocean helps to improve our understanding of the large-scale fate of DDTs in the Arctic.

Observation-Based Assessment of PBDE Loads in Arctic Ocean Waters.

Little is known about the distribution of polybrominated diphenyl ethers (PBDE) -also known as flame retardants- in major ocean compartments, with no reports yet for the large deep-water masses of the Arctic Ocean. Here, PBDE concentrations, congener patterns and inventories are presented for the different water masses of the pan-Arctic shelf seas and the interior basin. Seawater samples were collected onboard three cross-basin oceanographic campaigns in 2001, 2005, and 2008 following strict trace-clean protocols. ∑14PBDE concentrations in the Polar Mixed Layer (PML; a surface water mass) range from 0.3 to 11.2 pg·L(-1), with higher concentrations in the pan-Arctic shelf seas and lower levels in the interior basin. BDE-209 is the dominant congener in most of the pan-Arctic areas except for the ones close to North America, where penta-BDE and tetra-BDE congeners predominate. In deep-water masses, ∑14PBDE concentrations are up to 1 order of magnitude higher than in the PML. Whereas BDE-209 decreases with depth, the less-brominated congeners, particularly BDE-47 and BDE-99, increase down through the water column. Likewise, concentrations of BDE-71 -a congener not present in any PBDE commercial mixture- increase with depth, which potentially is the result of debromination of BDE-209. The inventories in the three water masses of the Central Arctic Basin (PML, intermediate Atlantic Water Layer, and the Arctic Deep Water Layer) are 158 ± 77 kg, 6320 ± 235 kg and 30800 ± 3100 kg, respectively. The total load of PBDEs in the entire Arctic Ocean shows that only a minor fraction of PBDEs emissions are transported to the Arctic Ocean. These findings represent the first PBDE data in the deep-water compartments of an ocean.

Serpentinization-associated travertines as spatio-temporal archives for lipid biomarkers key for the search for life on Mars.

Serpentinization is a well-known aqueous alteration process that may have played important roles in the origins and early evolution of life on Earth, and perhaps Mars, but there are still aspects related to biomarker distribution, partitioning, and preservation that merit further study. To assess the role that precipitation of carbonate phases in serpentinization settings may have on biomarker preservation, we search for life signs in one of the world's largest outcrops of subcontinental peridotites (Ronda, South Spain). We investigate the organic record of groundwater and associated carbonate deposits (travertines) in seven hyperalkaline springs, and reconstruct the biological activity and metabolic interactions of the serpentinization-hosted ecosystem. We identified lipid biomarkers and isotopic evidences of life, whose concentration and variety were much lower in groundwater than travertine deposits (ppb/ppt versus ppm level). Groundwater carried organics of abiotic (n-alkanes with values of CPI âˆ¼ 1) and/or biotic origin, of fresher (e.g. acids or alcohols) or more diagenetized (mature hopanes and n-alkanes) nature. In contrast, associated travertines held a more prolific record of biomarkers incorporating (molecular and isotopic) fingerprints of surface (mostly phototrophs) and subsurface (chemolithotrophs, methanogens and/or methanotrophs) life. Serpentinization-associated travertines seem to act as biomolecule archives over time fed by autochthonous and allochthonous sources, hence amplifying the dim biological signal of groundwater. These results illustrate the relevance of serpentinization-associated surface mineral deposits in searching for traces of life on analogous environments on Mars. We highlight the diversity of lipids produced in serpentinizing land environments and emphasize the potential of these geostable biomolecules to preserve fingerprints of life.

Lipid-based paleoecological and biogeochemical reconstruction of Store Saltsø, an extreme lacustrine system in SW Greenland.

Polar lakes harbour a unique biogeochemistry that reflects the implications of climatic fluctuations against a susceptible yet extreme environment. In addition to polar, Store Saltsø (Kangerlussuaq, southwestern Greenland) is an endorheic lake with alkaline and oligotrophic waters that host a distinctive ecology adapted to live in such particular physico-chemical and environmental conditions. By exploring the sedimentary record of Store Saltsø at a molecular and compound-specific isotopic level, we were able to understand its ecology and biogeochemical evolution upon climate change. We employed lipid biomarkers to identify biological sources and metabolic traits in different environmental samples (shore terrace, sediment core, and white precipitates at the shore), and their succession over time to reconstruct the lake paleobiology. Different molecular ratios and geochemical proxies provided further insights toward the evolution of environmental conditions in the frame of the deglaciation history of Kangerlussuaq. The relative abundance of terrestrial (i.e., plant derived) biomarkers (odd long-chain n-alkanes, even long-chain n-alkanols, and phytosterols) in the upper half of the shore terrace versus the relatively more present aquatic biomarkers (botryococcenes and long-chain alkenones) in its lower half revealed higher lake water levels in the past. Moreover, the virtual absence of organics in the deepest section of the sediment core (32-29 cm depth) suggested that the lake did not yet exist at the northwestern shore of Store Saltsø ∼5100 years ago. According to the relative abundance of lipid biomarkers detected in the adjacent section above (29-25 cm depth), we hypothesize that the northwestern shore of Store Saltsø formed ∼4900 years ago. By combining the molecular and compound-specific isotopic analysis of lipids in a ∼360 cm sedimentary sequence, we recreated the paleobiology and evolution of an extreme lacustrine environment suitable for the study of the limits of life and the effects of climate warming.

The Molecular Profile of Soil Microbial Communities Inhabiting a Cambrian Host Rock.

The process of soil genesis unfolds as pioneering microbial communities colonize mineral substrates, enriching them with biomolecules released from bedrock. The resultant intricate surface units emerge from a complex interplay among microbiota and plant communities. Under these conditions, host rocks undergo initial weathering through microbial activity, rendering them far from pristine and challenging the quest for biomarkers in ancient sedimentary rocks. In addressing this challenge, a comprehensive analysis utilizing Gas Chromatography Mass Spectrometry (GC-MS) and Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) was conducted on a 520-Ma-old Cambrian rock. This investigation revealed a diverse molecular assemblage with comprising alkanols, sterols, fatty acids, glycerolipids, wax esters, and nitrogen-bearing compounds. Notably, elevated levels of bacterial C16, C18 and C14 fatty acids, iso and anteiso methyl-branched fatty acids, as well as fungal sterols, long-chained fatty acids, and alcohols, consistently align with a consortium of bacteria and fungi accessing complex organic matter within a soil-type ecosystem. The prominence of bacterial and fungal lipids alongside maturity indicators denotes derivation from heterotrophic activity rather than ancient preservation or marine sources. Moreover, the identification of long-chain (>C22) n-alkanols, even-carbon-numbered long chain (>C20) fatty acids, and campesterol, as well as stigmastanol, provides confirmation of plant residue inputs. Furthermore, findings highlight the ability of contemporary soil microbiota to inhabit rocky substrates actively, requiring strict contamination controls when evaluating ancient molecular biosignatures or extraterrestrial materials collected.

Biosignature Detection and MinION Sequencing of Antarctic Cryptoendoliths After Exposure to Mars Simulation Conditions.

In the search for life in our Solar System, Mars remains a promising target based on its proximity and similarity to Earth. When Mars transitioned from a warmer, wetter climate to its current dry and freezing conditions, any putative extant life probably retreated into habitable refugia such as the subsurface or the interior of rocks. Terrestrial cryptoendolithic microorganisms (i.e., those inhabiting rock interiors) thus represent possible modern-day Mars analogs, particularly those from the hyperarid McMurdo Dry Valleys in Antarctica. As DNA is a strong definitive biosignature, given that there is no known abiotic chemistry that can polymerize nucleobases, we investigated DNA detection with MinION sequencing in Antarctic cryptoendoliths after an ∼58-sol exposure in MARTE, a Mars environmental chamber capable of simulating martian temperature, pressure, humidity, ultraviolet (UV) radiation, and atmospheric composition, in conjunction with protein and lipid detection. The MARTE conditions resulted in changes in community composition and DNA, proteins, and cell membrane-derived lipids remained detectable postexposure. Of the multitude of extreme environmental conditions on Mars, UV radiation (specifically UVC) is the most destructive to both cells and DNA. As such, we further investigated if a UVC exposure corresponding to ∼278 martian years would impede DNA detection via MinION sequencing. The MinION was able to successfully detect and sequence DNA after this UVC radiation exposure, suggesting its utility for life detection in future astrobiology missions focused on finding relatively recently exposed biomarkers inside possible martian refugia.

An Overview of Lipid Biomarkers in Terrestrial Extreme Environments with Relevance for Mars Exploration.

Lipid molecules are organic compounds, insoluble in water, and based on carbon-carbon chains that form an integral part of biological cell membranes. As such, lipids are ubiquitous in life on Earth, which is why they are considered useful biomarkers for life detection in terrestrial environments. These molecules display effective membrane-forming properties even under geochemically hostile conditions that challenge most of microbial life, which grants lipids a universal biomarker character suitable for life detection beyond Earth, where a putative biological membrane would also be required. What discriminates lipids from nucleic acids or proteins is their capacity to retain diagnostic information about their biological source in their recalcitrant hydrocarbon skeletons for thousands of millions of years, which is indispensable in the field of astrobiology given the time span that the geological ages of planetary bodies encompass. This work gathers studies that have employed lipid biomarker approaches for paleoenvironmental surveys and life detection purposes in terrestrial environments with extreme conditions: hydrothermal, hyperarid, hypersaline, and highly acidic, among others; all of which are analogous to current or past conditions on Mars. Although some of the compounds discussed in this review may be abiotically synthesized, we focus on those with a biological origin, namely lipid biomarkers. Therefore, along with appropriate complementary techniques such as bulk and compound-specific stable carbon isotope analysis, this work recapitulates and reevaluates the potential of lipid biomarkers as an additional, powerful tool to interrogate whether there is life on Mars, or if there ever was.

Ecological successions throughout the desiccation of Tirez lagoon (Spain) as an astrobiological time-analog for wet-to-dry transitions on Mars.

Tirez was a small and seasonal endorheic athalassohaline lagoon that was located in central Spain. In recent years, the lagoon has totally dried out, offering for the first time the opportunity to analyze its desiccation process as a "time-analog" to similar events occurred in paleolakes with varying salinity during the wet-to-dry transition on early Mars. On the martian cratered highlands, an early period of water ponding within enclosed basins evolved to a complete desiccation of the lakes, leading to deposition of evaporitic sequences during the Noachian and into the Late Hesperian. As Tirez also underwent a process of desiccation, here we describe (i) the microbial ecology of Tirez when the lagoon was still active 20 years ago, with prokaryotes adapted to extreme saline conditions; (ii) the composition of the microbial community in the dried lake sediments today, in many case groups that thrive in sediments of extreme environments; and (iii) the molecular and isotopic analysis of the lipid biomarkers that can be recovered from the sediments today. We discuss the implications of these results to better understanding the ecology of possible Martian microbial communities during the wet-to-dry transition at the end of the Hesperian, and how they may inform about research strategies to search for possible biomarkers in Mars after all the water was lost.

Assessing siliceous sinter matrices for long-term preservation of lipid biomarkers in opaline sinter deposits analogous to Mars in El Tatio (Chile).

Subaerial hydrothermal systems are of great interest for paleobiology and astrobiology as plausible candidate environments to support the origin of life on Earth that offer a unique and interrelated atmosphere-hydrosphere-lithosphere interface. They harbor extensive sinter deposits of high preservation potential that are promising targets in the search for traces of possible extraterrestrial life on Hesperian Mars. However, long-term quality preservation is paramount for recognizing biosignatures in old samples and there are still significant gaps in our understanding of the impact and extent of taphonomy processes on life fingerprints. Here, we propose a study based on lipid biomarkers -highly resistant cell-membrane components- to investigate the effects of silicification on their preservation in hydrothermal opaline sinter. We explore the lipid biomarkers profile in three sinter deposits of up to ~3000 years from El Tatio, one of the best Martian analogs on Earth. The lipid profile in local living biofilms is used as a fresh counterpart of the fossil biomarkers in the centuries-old sinter deposits to qualitatively assess the taphonomy effects of silicification on the lipid's preservation. Despite the geological alteration, the preserved lipids retained a depleted stable-carbon isotopic fingerprint characteristic of biological sources, result highly relevant for astrobiology. The data allowed us to estimate for the first time the degradation rate of lipid biomarkers in sinter deposits from El Tatio, and to assess the time preservation framework of opaline silica. Auxiliary techniques of higher taxonomic resolution (DNA sequencing and metaproteomics) helped in the reconstruction of the paleobiology. The lipids were the best-preserved biomolecules, whereas the detection of DNA and proteins dropped considerably from 5 cm depth. These findings provide new insights into taphonomy processes affecting life fingerprints in hydrothermal deposits and serves as a useful baseline for assessing the time window for recovering unambiguous signs of past life on Earth and beyond.

Life Detection and Microbial Biomarker Profiling with Signs of Life Detector-Life Detector Chip During a Mars Drilling Simulation Campaign in the Hyperarid Core of the Atacama Desert.

The low organic matter content in the hyperarid core of the Atacama Desert, together with abrupt temperature shifts and high ultraviolet radiation at its surface, makes this region one of the best terrestrial analogs of Mars and one of the best scenarios for testing instrumentation devoted to in situ planetary exploration. We have operated remotely and autonomously the SOLID-LDChip (Signs of Life Detector-Life Detector Chip), an antibody microarray-based sensor instrument, as part of a rover payload during the 2019 NASA Atacama Rover Astrobiology Drilling Studies (ARADS) Mars drilling simulation campaign. A robotic arm collected drilled cuttings down to 80 cm depth and loaded SOLID to process and assay them with LDChip for searching for molecular biomarkers. A remote science team received and analyzed telemetry data and LDChip results. The data revealed the presence of microbial markers from Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria, Firmicutes, and Cyanobacteria to be relatively more abundant in the middle layer (40-50 cm). In addition, the detection of several proteins from nitrogen metabolism indicates a pivotal role in the system. These findings were corroborated and complemented on "returned samples" to the lab by a comprehensive analysis that included DNA sequencing, metaproteomics, and a metabolic reconstruction of the sampled area. Altogether, the results describe a relatively complex microbial community with members capable of nitrogen fixation and denitrification, sulfur oxidation and reduction, or triggering oxidative stress responses, among other traits. This remote operation demonstrated the high maturity of SOLID-LDChip as a powerful tool for remote in situ life detection for future missions in the Solar System.

Dark microbiome and extremely low organics in Atacama fossil delta unveil Mars life detection limits.

Identifying unequivocal signs of life on Mars is one of the most important objectives for sending missions to the red planet. Here we report Red Stone, a 163-100 My alluvial fan-fan delta that formed under arid conditions in the Atacama Desert, rich in hematite and mudstones containing clays such as vermiculite and smectites, and therefore geologically analogous to Mars. We show that Red Stone samples display an important number of microorganisms with an unusual high rate of phylogenetic indeterminacy, what we refer to as "dark microbiome", and a mix of biosignatures from extant and ancient microorganisms that can be barely detected with state-of-the-art laboratory equipment. Our analyses by testbed instruments that are on or will be sent to Mars unveil that although the mineralogy of Red Stone matches that detected by ground-based instruments on the red planet, similarly low levels of organics will be hard, if not impossible to detect in Martian rocks depending on the instrument and technique used. Our results stress the importance in returning samples to Earth for conclusively addressing whether life ever existed on Mars.

Ecological variability based on lipid biomarkers in astrobiologically interesting wetlands from the Argentinian central Andes.

Andean wetlands hold extremophilic communities adapted to live in harsh conditions. Here, we investigated the microbial ecology of three high-altitude hypersaline ponds from La Puna region (Argentina) showing an increasing extent of desiccation by analyzing their lipid sedimentary record. We recreated the microbial community structure and the carbon metabolisms in each lacustrine system based on the molecular distribution of lipid biomarkers and their compound-specific carbon and hydrogen isotopic signatures. We detected lipid compounds considered to be biomarkers of cyanobacteria, sulfate-reducing bacteria, purple sulfur bacteria, and archaea in the three Andean ponds, as well as diatoms in the intermediate salinity system. The relative abundance of purple sulfur and sulfate-reducing bacteria decreased with salinity, whereas cyanobacteria and archaea decreased their relative abundance in the mid-saline pond to increase it again and became both prevailing at the highest salinity. Carbon fixation in the three ponds was driven by a combination of the reductive tricarboxylic acid cycle, the reductive pentose phosphate cycle, and the reductive acetyl-CoA pathway. This work is the first to describe molecular and isotopic lipid fingerprints in wetlands from the central Andean Puna, and serves as a basis for further biogeochemical studies in the area.

Diversity and Effect of Increasing Temperature on the Activity of Methanotrophs in Sediments of Fildes Peninsula Freshwater Lakes, King George Island, Antarctica.

Global warming has a strong impact on polar regions. Particularly, the Antarctic Peninsula and nearby islands have experienced a marked warming trend in the past 50 years. Therefore, higher methane (CH4) emissions from this area could be expected in the future. Since mitigation of these emissions can be carried out by microbial oxidation, understanding this biological process is crucial since to our knowledge, no related studies have been performed in this area before. In this work, the aerobic CH4 oxidation potential of five freshwater lake sediments of Fildes Peninsula (King George Island, South Shetland Islands) was determined with values from 0.07 to 10 µmol CH4 gdw-1 day-1 and revealed up to 100-fold increase in temperature gradients (5, 10, 15, and 20°C). The structure and diversity of the bacterial community in the sediments were analyzed by next-generation sequencing (Illumina MiSeq) of 16S rRNA and pmoA genes. A total of 4,836 ASVs were identified being Proteobacteria, Actinobacteriota, Acidobacteriota, and Bacteroidota the most abundant phyla. The analysis of the pmoA gene identified 200 ASVs of methanotrophs, being Methylobacter Clade 2 (Type I, family Methylococcaceae) the main responsible of the aerobic CH4 oxidation. Moreover, both approaches revealed the presence of methanotrophs of the classes Gammaproteobacteria (families Methylococcaceae and Crenotrichaceae), Alphaproteobacteria (family Methylocystaceae), Verrucomicrobia (family Methylacidiphilaceae), and the candidate phylum of anaerobic methanotrophs Methylomirabilota. In addition, bacterial phospholipid fatty acids (PLFA) biomarkers were studied as a proxy for aerobic methane-oxidizing bacteria and confirmed these results. Methanotrophic bacterial diversity was significantly correlated with pH. In conclusion, our findings suggest that aerobic methanotrophs could mitigate in situ CH4 emissions in a future scenario with higher temperatures in this climate-sensitive area. This study provides new insights into the diversity of methanotrophs, as well as the influence of temperature on the CH4 oxidation potential in sediments of freshwater lakes in polar regions of the southern hemisphere.

Raman spectroscopic peculiarities of Icelandic poorly crystalline minerals and their implications for Mars exploration.

In this work, we have analyzed natural samples collected at three hydrothermal areas of Iceland by Raman spectroscopy. The studied high-latitude regions are considered environmentally and mineralogically appropriate Martian analogues since they are rich in weathered basalts that have been altered by hydrothermalism to mineral phases such as silica, clay minerals, sulfates, oxides, and sulfur. The main objective of this work was to assess the relation of the spectroscopic signatures of alteration to hydrothermal processes and biomediation, considering previous studies focused on the detection of lipid biomarkers in the same samples. The recorded Raman spectra, taken with optical parameters similar to the ExoMars 2022 Raman spectrometer, showed structural modifications in all secondary minerals in the form of peak shifts (in the case of sulfur and clay minerals), changes in the relative ratio intensity (in anatase) and/or shape broadening (in sulfates and hematite). These results reveal the suitability of Raman spectroscopy to examine areas rich in water-altered minerals, where a mixture of crystalline and amorphous phases can co-exist. The detection of silica is singularly interesting since, on the one hand, it can imply the past existence of hydrothermal hot springs rich in nutrient and redox gradients and, on the other hand, provides excellent matrix for biosignature preservation. The data can be helpful as an astrobiological database for the forthcoming missions to Mars, where potential upwelling groundwater systems could have altered the mineral phases in a similar way to that observed in this work.

Biospeleothems Formed by Fungal Activity During the Early Holocene in the "Salar de Uyuni".

The Chiquini and Galaxias caves contain speleothems that are templated by long fungal structures. They have been associated with the carbonate lacustrine deposits in the margins of the Coipasa and Uyuni Salar basins. During a wetter episode, such carbonates formed at the end of the last glaciation raising the lake level to more than 100 m in the Tauca events (15-12 ky). Such an event flooded the caves that eventually became a cryptic habitat in the lake. The caves show bizarre speleothems framed by large (>1 m) fungal buildings covering the older algal mineralized structures. Although the origin of the caves is not fully understood, the occurrence of two carbonatic units with very distinctive fabric suggests that they formed in two separated humid events. In this regard, the mineralized algal structures, showing the same features as the lacustrine carbonates, likely formed during the Tauca flooding events in the terminal Pleistocene that inundated older caves. The different caves were exposed to the atmosphere after a drop in the lake level that promoted alluvial erosion by <12-10 ky (Ticaña episode) under arid conditions. A last humid episode rising the lake surface 10 m above the Salar level, which was not enough to inundate the caves a second time, drove the formation of the biospeleothems by fungi biomineralization. The abundance and size of the preserved fungal structures suggest that they were sustained by a stable hydrological activity plus a constant organic supply. While nutrients could have been primarily sourced from the vegetal communities that occupied the exhumated lake margins, they might have also been released from the lacustrine carbonatic unit. The combination of hydrology and biological activities were likely determinants for a fast rock dissolution and mineralization ending in the construction of the fungal biospeleothems.