The differentiation between living and dead bacterial endospores is crucial in many research areas of microbiology. The identification of inactivated, non-pathogenic Bacillus anthracis spores is one reason why improvement of decontamination protocols is so desirable. Another field interested in spore viability is planetary protection, a sub-discipline of astrobiology that estimates the bioburden of spacecraft prior to launch in order to avoid interplanetary cross-contamination. We developed a dedicated, rapid and cost-effective method for identifying bacterial endospores that have been inactivated and consequently show a compromised spore wall. This novel protocol is culture-independent and is based on fluorescence microscopy and propidium monoazide (PMA) as a fluorescent marker, which is suggested to bind to DNA of spores with compromised spore coat, cortex and membranes based on our results. Inactivated preparations (treated with wet heat, irradiation, ultracentrifugation) showed a significant increase in spores that were PMA stained in their core; moreover, Bacillus atrophaeus, Bacillus safensis and Geobacillus stearothermophilus seemed to be best suited for this technique, as the spore cores of all these endospores could be positively stained after inactivation. Lastly, we describe an additional counter-staining protocol and provide an example of the application of the coupled staining methods for planetary protection purposes. The introduction of this novel protocol is expected to provide an initial insight into the various possible future applications of PMA as a non-viability marker for spores in, for example, B. anthracis-related studies, food microbiology and astrobiology.

Raman spectrometry has been established as an instrument of choice for studying the structure and bond type of known molecules, and identifying the composition of unknown substances, whether geological or biological. This versatility has led to its strong consideration for planetary exploration. In the context of the ExoGeoLab and ExoHab pilot projects of ESA-ESTEC & ILEWG (International Lunar Exploration Working Group), we investigated samples of astrobiological interest using a portable Raman spectrometer lasing at 785 nm and discuss implications for planetary exploration. We find that biological samples are typically best observed at wavenumbers >1100 cm−1, but their Raman signals are often affected by fluorescence effects, which lowers their signal-to-noise ratio. Raman signals of minerals are typically found at wavenumbers <1100 cm−1, and tend to be less affected by fluorescence. While higher power and/or longer signal integration time improve Raman signals, such power settings are detrimental to biological samples due to sample thermal degradation. Care must be taken in selecting the laser wavelength, power level and integration time for unknown samples, particularly if Raman signatures of biological components are anticipated. We include in the Appendices tables of Raman signatures for astrobiologically relevant organic compounds and minerals.

Synchrotron radiation-based spectroscopic techniques are discussed. Their relevance to obtain information regarding the prebiotic evolution problem is pointed out. We present photoelectron–photoion coincidence (PEPICO) spectra of adenine and glycine obtained using 12 and 21 eV photons. The fragmentation pattern belonging to these molecules was found to present striking differences, which are discussed. Adenine partial ion yield in the energy region 12–21 eV is also presented. The neutral fragments were found to have very simple assignment. The importance of hydrogen cyanide (HCN) as a building block of these molecules is confirmed. A special instrumentation allowing precise comparisons between photon-induced desorption and energetic ion bombardment desorption is described. As an example, we show, for the first time, the frozen CO2 ice mass spectra bombarded by photons and energetic ions, under the same experimental conditions. The comparison shows that prebiotic evolution may only be properly understood if more than one particle, as energy source, is considered.

Most chemical reactions on asteroids, from which meteors and meteorites originate, are hypothesized to occur primarily in the solid mixtures. Some secondary chemical reactions may have occurred during the periods of the aqueous alteration of the asteroids. A myriad of organic compounds have been isolated from the meteorites, but the chemical conditions during which they were formed are only partially elucidated. In this paper, we propose that numerous meteoritic organic compounds were formed by the solventless and solid-state reactions that were only recently explored in conjunction with the green chemistry. A typical solventless approach exploits the phenomenon of the mixed melting points. As the solid materials are mixed together, the melting point of the mixture becomes lower than the melting points of its individual components. In some cases, the entire mixture may melt upon mixing. These reactions could then occur in a melted state. In the traditional solid-state reactions, the solids are mixed together, which allows for the intimate contact of the reactants, but the reaction occurs without melting. We have shown various examples of the known solventless and solid-state reactions that are particularly relevant to the meteoritic chemistry. We have also placed them in a prebiotic context and evaluated them for their astrobiological significance.

Nuclear astrophysics is a relatively young science; it is about half a century old. It is a multidisciplinary subject, since it combines nuclear physics with astrophysics and observations in astronomy. It also addresses fundamental issues in astrobiology through the formation of elements, in particular those required for a carbon-based life. In this paper, a rapid overview of nucleosynthesis is given, mainly from the point of view of nuclear physics. A short historical introduction is followed by the definition of the relevant nuclear parameters, such as nuclear reaction cross sections, astrophysical S-factors, the energy range defined by the Gamow peak and reaction rates. The different astrophysical scenarios that are the sites of nucleosynthesis, and different processes, cycles and chains that are responsible for the building of complex nuclei from the elementary hydrogen nuclei are then briefly described.

For over 50 years, astronomers have searched the skies for evidence of electromagnetic signals from extraterrestrial (ET) civilizations that have reached or surpassed our level of technological development. Although often overlooked or given as granted, the parallel use of an equivalent communication technology is a necessary prerequisite for establishing contact in both leakage and deliberate messaging strategies. Civilization advancements, especially accelerating change and exponential growth, lessen the perspective for a simultaneous technological status of civilizations thus putting hard constraints on the likelihood of a dialogue. In this paper, we consider the mathematical probability of technological synchronicity of our own and a number of other hypothetical ET civilizations and explore the most likely scenarios for their concurrency. If Search for Extraterrestrial Intelligence (SETI) projects rely on a fortuitous detection of leaked interstellar signals (so-called ‘eavesdropping’) then with minimum prior assumptions N⩾138–4991 Earth-like civilizations have to exist at this moment in the Galaxy for the technological usage synchronicity probability p⩾0.95 in the next 20 years. We also show that since the emergence of complex life, coherent with the hypothesis of the Galactic habitable zone (GHZ), N⩾1497 ET civilizations had to be created in the Galaxy in order to achieve the same estimated probability in the technological possession synchronicity that corresponds to the deliberate signalling scenario.

Planetary protection regulations are in place to control the contamination of planets and moons with terrestrial micro-organisms in order to avoid jeopardizing future scientific investigations relating to the search for life. One environmental chemical factor of relevance in extraterrestrial environments, specifically in the moons of the outer solar system, is ammonia (NH3). Ammonia is known to be highly toxic to micro-organisms and may disrupt proton motive force, interfere with cellular redox reactions or cause an increase of cell pH. To test the survival potential of terrestrial micro-organisms exposed to such cold, ammonia-rich environments, and to judge whether current planetary protection regulations are sufficient, soil samples were exposed to concentrations of NH3 from 5 to 35% (v/v) at −80°C and room temperature for periods up to 11 months. Following exposure to 35% NH3, diverse spore-forming taxa survived, including representatives of the Firmicutes (Bacillus, Sporosarcina, Viridibacillus, Paenibacillus, Staphylococcus and Brevibacillus) and Actinobacteria (Streptomyces). Non-spore forming organisms also survived, including Proteobacteria (Pseudomonas) and Actinobacteria (Arthrobacter) that are known to have environmentally resistant resting states. Clostridium spp. were isolated from the exposed soil under anaerobic culture. High NH3 was shown to cause a reduction in viability of spores over time, but spore morphology was not visibly altered. In addition to its implications for planetary protection, these data show that a large number of bacteria, potentially including spore-forming pathogens, but also environmentally resistant non-spore-formers, can survive high ammonia concentrations.

Although the field of synthetic biology is still in its infancy, there are expectations for great advances in the coming decades, both on Earth and potentially in space. Promising applications for long duration space missions include a variety of biologically engineered products and biologically aided processes and technologies, which will undoubtedly be scrutinized for risks and benefits in the broad context of ethical, legal and social realms. By comparing and contrasting features of Earth-based and space-applied synthetic biology, it is possible to identify the likely similarities and differences, and to identify possible challenges ahead for space applications that will require additional research, both in the short and long terms. Using an analytical framework associated with synthetic biology and new technologies on Earth, this paper analyses the kinds of issues and concerns ahead, and identifies those areas where space applications may require additional examination. In general, while Earth- and space-based synthetic biology share many commonalities, space applications have additional challenges such as those raised by space microbiology and environmental factors, legal complications, planetary protection, lack of decision-making infrastructure(s), long duration human missions, terraforming and the possible discovery of extraterrestrial (ET) life. For synthetic biology, the way forward offers many exciting opportunities, but is not without legitimate concerns – for life, environments and society, both on Earth and beyond.

Astrobiology is a relatively recent scientific field that seeks to understand the origin and dynamics of life in the Universe. Several hypotheses have been proposed to explain life in the cosmic context throughout human history, but only now, technology has allowed many of them to be tested. Laboratory experiments have been able to show how chemical elements essential to life, such as carbon, nitrogen, oxygen and hydrogen combine in biologically important compounds. Interestingly, these compounds are ubiquitous. How these compounds were combined to the point of originating cells and complex organisms is still to be unveiled by science. However, our 4.5 billion years old Solar system appeared in a 10 billion years old Universe. Thus, simple cells such as micro-organisms may have had time to form in planets older than ours or in other suitable places in the Universe. One hypothesis related to the appearance of life on Earth is called panspermia, which predicts that microbial life could have been formed in the Universe billions of years ago, travelling between planets, and inseminating units of life that could have become more complex in habitable planets such as Earth. A project designed to test the viability of extremophile micro-organisms exposed to simulated extraterrestrial environments is in progress at the Carlos Chagas Filho Institute of Biophysics (UFRJ, Brazil) to test whether microbial life could withstand inhospitable environments. Radiation-resistant (known or novel ones) micro-organisms collected from extreme terrestrial environments have been exposed (at synchrotron accelerators) to intense radiation sources simulating Solar radiation, capable of emitting radiation in a few hours equivalent to many years of accumulated doses. The results obtained in these experiments reveal an interesting possibility of the existence of microbial life beyond Earth.

A high research priority in astrobiology is the search and eventual identification of biomarkers in the Solar System. In spite of numerous steps forward, lunar science remains largely disjoint from the main stream of astrobiology, but in recent years the Moon has begun to emerge as a novel target for astrobiologists. We discuss an overlap between lunar geology and terrestrial geomicrobiology that arises from analysis of lunar soils and some uncertainties in chemical evolution and the origin of life scenarios. Unexpected isotopic heterogeneity of nitrogen (N) was found to be remarkable in samples from Apollo and the Luna programme. Both the stable isotope geochemical data of the biogenic elements, as well as the noble gases trapped in lunar soils added valuable new and relevant data. These discoveries are potential sources of information on early Earth evolution. The elusive ratio of N's two stable isotopes 15N/14N has played a fundamental role in this research. The analysis of individual grains of ilmenite suggests that 90% of all the trapped N does not originate from solar wind (SW). We discuss the significance of these stable isotopes from the point of view of astrobiology in the light of the next generation of lunar exploration. We underline the high priority of testing the origin of non-solar N source trapped in the regolith of the lunar farside. In the proposals of new lunar missions, the characterization of the geochemistry at several lunar sites is a major objective. Some arguments are presented in favour of using novel space technologies in a search for biomarkers in geographical distinct lunar landing sites. We restrict our attention to one aspect of the science requirements for the forthcoming missions by focusing on a very limited objective: to take a closer look at the geochemical characterization of the chemical element N on the soils of the lunar farside.

Life inhabits the subsurface of the Earth down to depths where temperature precludes it. Similar conditions are likely to exist within the traditional habitable zone for objects between 0.1 Earth mass (Mars) and 10 Earth masses (superearth). Long-term cooling and internal radioactivity maintain surface heat flow on the Earth. These heat sources are comparable and likely to be comparable in general within old rocky planets. Surface heat flow scales with mass divided by surface area and hence with surface gravity. The average absolute habitable subsurface thickness scales inversely with heat flow and gravity. Surface gravity varies by only 0.4 g for Mars to 3.15 g for a superearth. This range is less than the regional variation of heat flow on the Earth. Still ocean-boiling asteroid impacts (if they occur) are more likely to sterilize the thin habitable subsurface of large objects than thick habitable subsurface of small ones. Tectonics self-organizes to maintain subsurface permeability and habitability within both stable and active regions on the Earth. Small earthquakes within stable regions allow sudden mixing of water masses. Large earthquakes at plate boundaries allow surface water to descend to great habitable depths. Seismic shaking near major faults cracks shallow rock forming permeable regolith. Strong tidal strains form a similar porous regolith on small bodies such as Enceladus with weak stellar heating. This regolith may be water-saturated within rocky bodies and thus habitable.

The survival strategies of extremophilic organisms in terrestrially stressed locations and habitats are critically dependent on the production of protective chemicals in response to desiccation, low wavelength radiation insolation, temperature and the availability of nutrients. The adaptation of life to these harsh prevailing conditions involves the control of the substratal geology; the interaction between the rock and the organisms is critical and the biological modification of the geological matrix plays a very significant role in the overall survival strategy. Identification of these biological and biogeological chemical molecular signatures in the geological record is necessary for the recognition of the presence of extinct or extant life in terrestrial and extraterrestrial scenarios. Raman spectroscopic techniques have been identified as valuable instrumentation for the detection of life extra-terrestrially because of the use of non-invasive laser-based excitation of organic and inorganic molecules, and molecular ions with high discrimination characteristics; the interactions effected between biological organisms and their environments are detectable through the molecular entities produced at the interfaces, for which the vibrational spectroscopic band signatures are unique. A very important attribute of Raman spectroscopy is the acquisition of molecular experimental data non-destructively without the need for chemical or mechanical pre-treatment of the specimen; this has been a major factor in the proposal for the adoption of Raman instrumentation on robotic landers and rovers for planetary exploration, particularly for the forthcoming European Space Agency (ESA)/National Aeronautics and Space Administration (NASA) ExoMars mission. In this paper, the merits of using Raman spectroscopy for the recognition of key molecular biosignatures from several terrestrial extremophile specimens will be illustrated. The data and specimens used in this presentation have been acquired from Arctic and Antarctic cold deserts and a meteorite crater, from which it will be possible to assess spectral data relevant for the detection of extra-terrestrial extremophilic life signatures.

With the possible exception of meteor impacts, high-energy astrophysical events such as supernovae, gamma-ray bursts (GRB) and flares are usually not taken into account for biological and evolutionary studies due to their low rates of occurrence. We show that a class of these events may occur at distances and time scales in which their biological effects are non-negligible, maybe more frequent than the impacts of large asteroids. We review the effects of four transient astrophysical sources of ionizing radiation on biospheres – stellar flares, giant flares from soft gamma repeaters (SGR), supernovae and GRB. The main damaging features of them are briefly discussed and illustrated. We point out some open problems and ongoing work.

A key open question in the study of life is the origin of biomolecular homochirality: almost every life-form on Earth has exclusively levorotary amino acids and dextrorotary sugars. Will the same handedness be preferred if life is found elsewhere? We review some of the pertinent literature and discuss recent results suggesting that life's homochirality resulted from sequential chiral symmetry breaking triggered by environmental events. In one scenario, autocatalytic prebiotic reactions undergo stochastic fluctuations due to environmental disturbances, in a mechanism reminiscent of evolutionary punctuated equilibrium: short-lived destructive events may lead to long-term enantiomeric excess. In another, chiral-selective polymerization reaction rates influenced by environmental effects lead to substantial chiral excess even in the absence of autocatalysis. Applying these arguments to other potentially life-bearing platforms has implications to the search for extraterrestrial life: we predict that a statistically representative sampling of extraterrestrial stereochemistry will be racemic (chirally neutral) on average.