Exobiology of the Venusian Clouds: New Insights into Habitability through Terrestrial Models and Methods of Detection.

The search for life beyond Earth has focused on Mars and the icy moons Europa and Enceladus, all of which are considered a safe haven for life due to evidence of current or past water. The surface of Venus, on the other hand, has extreme conditions that make it a nonhabitable environment to life as we know it. This is in contrast, however, to its cloud layer, which, while still an extreme environment, may prove to be a safe haven for some extreme forms of life similar to extremophiles on Earth. We consider the venusian clouds a habitable environment based on the presence of (1) a solvent for biochemical reactions, (2) appropriate physicochemical conditions, (3) available energy, and (4) biologically relevant elements. The diversity of extreme microbial ecosystems on Earth has allowed us to identify terrestrial chemolithoautotrophic microorganisms that may be analogs to putative venusian organisms. Here, we hypothesize and describe biological processes that may be performed by such organisms in the venusian clouds. To detect putative venusian organisms, we describe potential biosignature detection methods, which include metal-microbial interactions and optical methods. Finally, we describe currently available technology that can potentially be used for modeling and simulation experiments.

Effects of Radiation Intensity, Mineral Matrix, and Pre-Irradiation on the Bacterial Resistance to Gamma Irradiation under Low Temperature Conditions.

Ionizing radiation is one of the main factors limiting the survival of microorganisms in extraterrestrial conditions. The survivability of microorganisms under irradiation depends significantly on the conditions, in which the irradiation occurs. In particular, temperature, pressure, oxygen and water concentrations are of great influence. However, the influence of factors such as the radiation intensity (in low-temperature conditions) and the type of mineral matrix, in which microorganisms are located, has been practically unstudied. It has been shown that the radioresistance of bacteria can increase after their exposure to sublethal doses and subsequent repair of damage under favorable conditions, however, such studies are also few and the influence of other factors of extraterrestrial space (temperature, pressure) was not studied in them. The viability of bacteria Arthrobacter polychromogenes, Kocuria rosea and Xanthomonas sp. after irradiation with gamma radiation at a dose of 1 kGy under conditions of low pressure (1 Torr) and low temperature (-50 °C) at different radiation intensities (4 vs. 0.8 kGy/h) with immobilization of bacteria on various mineral matrices (montmorillonite vs. analogue of lunar dust) has been studied. Native, previously non-irradiated strains, and strains that were previously irradiated with gamma radiation and subjected to 10 passages of cultivation on solid media were irradiated. The number of survived cells was determined by culturing on a solid medium. It has been shown that the radioresistance of bacteria depends significantly on the type of mineral matrix, on which they are immobilized, wherein montmorillonite contributes to an increased survivability in comparison with a silicate matrix. Survivability of the studied bacteria was found to increase with decreasing radiation intensity, despite the impossibility of active reparation processes under experimental conditions. Considering the low intensity of radiation on various space objects in comparison with radiobiological experiments, this suggests a longer preservation of the viable microorganisms outside the Earth than is commonly believed. An increase in bacterial radioresistance was revealed even after one cycle of irradiation of the strains and their subsequent cultivation under favourable conditions. This indicates the possibility of hypothetical microorganisms on Mars increasing their radioresistance.

Phobos LIFE (Living Interplanetary Flight Experiment).

The Planetary Society's Phobos Living Interplanetary Flight Experiment (Phobos LIFE) flew in the sample return capsule of the Russian Federal Space Agency's Phobos Grunt mission and was to have been a test of one aspect of the hypothesis that life can move between nearby planets within ejected rocks. Although the Phobos Grunt mission failed, we present here the scientific and engineering design and motivation of the Phobos LIFE experiment to assist with the scientific and engineering design of similar future experiments. Phobos LIFE flew selected organisms in a simulated meteoroid. The 34-month voyage would have been the first such test to occur in the high-radiation environment outside the protection of Earth's magnetosphere for more than a few days. The patented Phobos LIFE "biomodule" is an 88 g cylinder consisting of a titanium outer shell, several types of redundant seals, and 31 individual Delrin sample containers. Phobos LIFE contained 10 different organisms, representing all three domains of life, and one soil sample. The organisms are all very well characterized, most with sequenced genomes. Most are extremophiles, and most have flown in low Earth orbit. Upon return from space, the health and characteristics of organisms were to have been compared with controls that remained on Earth and have not yet been opened.

Microbial activity in Martian analog soils after ionizing radiation: implications for the preservation of subsurface life on Mars.

At present, the surface of Mars is affected by a set of factors that can prevent the survival of Earth-like life. However, the modern concept of the evolution of the planet assumes the existence more favorable for life climate in the past. If in the past on Mars had formed a biosphere, similar to the one that originated in the early Earth, it is supposed that it is preserved till now in anabiotic state in the bowels of the planet, like microbial communities inhabiting the ancient permafrost of Arctic and Antarctic. In the conditions of modern Martian regolith, this relic life seems to be deprived of the possibility of damage reparation (or these processes occur on a geological time scale), and ionizing radiation should be considered the main factor inhibiting such anabiotic life. In the present study, we studied soil samples, selected in two different extreme habitats of the Earth: ancient permafrost from the Dry Valleys of Antarctica and Xerosol soil from the mountain desert in Morocco, gamma-irradiated with 40 kGy dose at low pressure (1 Torr) and low temperature (-50 °C). Microbial communities inhabiting these samples showed in situ high resistance to the applied effects, retained high number of viable cells, metabolic activity, and high biodiversity. Based on the results, it is assumed that the putative biosphere could be preserved in the dormant state for at least 500 thousand years and 8 million years in the surface layer of Mars regolith and at 5 m depth, respectively, at the current level of ionizing radiation intensity.

Soil bacterial communities of Sahara and Gibson deserts: Physiological and taxonomical characteristics.

The purpose of this research was to investigate the structure of soil bacteria communities present in the Gibson (Australia) and the Sahara (Egypt) deserts, as well as to estimate strain survivability under different environmental factors. It should be noticed that the screening of bacterial resistance to wide spectra of principally different stress conditions was performed for the first time. Experiments were conducted with culturable bacterial communities. Strains were identified using 16S rRNA sequencing, and stress-tolerance was estimated by growing strains in various nutrient media. In order to characterize the community the epifluorescent microscopy and multisubstrate testing were also performed. High bacterial abundance in the desert soils was detected, and there was seen a significant proportion of culturable cells. The close numbers of psychotropic and mesophilic bacteria in arid ecosystems were revealed. The representatives of the Actinobacteria phylum were dominant in the microbial communities, and Firmicutes, Proteobacteria, and Bacteroidetes phyla representatives were also identified. Tolerance of the axenic bacterial cultures, isolated from arid desert ecotopes, to temperature, pH, salts (KCl, NaCl, MgSO4, NaHCO3), strong oxidizers (Mg(ClO4)2), and antibiotics (ampicillin, cephalexin, chloramphenicol, tetracycline, doxycycline, kanamycin, rifampicin) was studied. The bacterial isolates were characterized by polyextremotolerance and by the ability to maintain metabolic activity in vitro while influenced by a wide range of physicochemical and biotic factors.

100 kGy gamma-affected microbial communities within the ancient Arctic permafrost under simulated Martian conditions.

This research aimed to investigate the viability and biodiversity of microbial communities within ancient Arctic permafrost after exposure to a gamma-radiation dose of 100 kGy at low temperature (- 50 °C), low pressure (1 Torr) and dehydration conditions. The main objective was to assess the possibility for long-term survival of Earth-bound microorganisms in the subsurface of Martian regolith or inside small space bodies at constant absorption and accumulation of the gamma radiation dose. Investigated microbial communities had shown high resistance to a simulated Martian environment. After irradiation the total count of prokaryotic cells and number of metabolically active bacterial cells remained at the control level, while the number of bacterial CFUs decreased by 2 orders of magnitude, and the number of metabolically active cells of archaea decreased threefold. Besides, the abundance of culturable bacteria after irradiation was kept at a high level: not less than 3.7 × 105 cells/g. Potential metabolic activity of irradiated microbial communities in general were higher than in the control sample. A fairly high biodiversity of bacteria was detected in the exposed sample of permafrost, although the microbial community structure underwent significant changes after irradiation. In particular, actinobacteria populations of the genus Arthrobacter, which was not revealed in the control samples, became predominant in bacterial communities following the exposure. The results of the study testify that long-term preservation of microbial life inside Martian permafrost is possible. The data obtained can also be evaluated from the perspective of the potential for discovering viable Earth-bound microorganisms on other objects in the Solar system and inside of small bodies in outer space.

The structure of resting bacterial populations in soil and subsoil permafrost.

The structure of individual cells in microbial populations in situ of the Arctic and Antarctic permafrost was studied by scanning and transmission electron microscopy methods and compared with that of cyst-like resting forms generated under special conditions by the non-spore-forming bacteria Arthrobacter and Micrococcus isolated from the permafrost. Electron microscopy examination of microorganisms in situ revealed several types of bacterial cells having no signs of damage, including "dwarf" curved forms similar to nanoforms. Intact bacterial cells in situ and frozen cultures of the permafrost isolates differed from vegetative cells by thickened cell walls, the altered structure of cytoplasm, and the compact nucleoid, and were similar in these features to cyst-like resting forms of non-spore-forming "permafrost" bacterial strains of Arthrobacter and Micrococcus spp. Cyst-like cells, being resistant to adverse external factors, are regarded as being responsible for survival of the non-spore-formers under prolonged exposure to subzero temperatures and can be a target to search for living microorganisms in natural environments both on the Earth and on extraterrestrial bodies.