Bacillus subtilis spore survival and expression of germination-induced bioluminescence after prolonged incubation under simulated Mars atmospheric pressure and composition: implications for planetary protection and lithopanspermia

Bacterial endospores in the genus Bacillus are considered good models for studying interplanetary transfer of microbes by natural or human processes. Although spore survival during transfer itself has been the subject of considerable study, the fate of spores in extraterrestrial environments has received less attention. In this report we subjected spores of a strain of Bacillus subtilis, containing luciferase resulting from expression of an sspB-luxAB gene fusion, to simulated martian atmospheric pressure (7-18 mbar) and composition (100% CO(2)) for up to 19 days in a Mars simulation chamber. We report here that survival was similar between spores exposed to Earth conditions and spores exposed up to 19 days to simulated martian conditions. However, germination-induced bioluminescence was lower in spores exposed to simulated martian atmosphere, which suggests sublethal impairment of some endogenous spore germination processes.     

Survival of spores of the UV-resistant Bacillus subtilis strain MW01 after exposure to low-earth orbit and simulated martian conditions: data from the space experiment ADAPT on EXPOSE-E

In the space experiment "Molecular adaptation strategies of microorganisms to different space and planetary UV climate conditions" (ADAPT), bacterial endospores of the highly UV-resistant Bacillus subtilis strain MW01 were exposed to low-Earth orbit (LEO) and simulated martian surface conditions for 559 days on board the European Space Agency's exposure facility EXPOSE-E, mounted outside the International Space Station. The survival of B. subtilis MW01 spores from both assays (LEO and simulated martian conditions) was determined by a colony-formation assay after retrieval. It was clearly shown that solar extraterrestrial UV radiation (λ≥110?nm) as well as the martian UV spectrum (λ≥200?nm) was the most deleterious factor applied; in some samples only a few spore survivors were recovered from B. subtilis MW01 spores exposed in monolayers. However, if shielded from solar irradiation, about 8% of MW01 spores survived in LEO conditions, and 100% survived in simulated martian conditions, compared to the laboratory controls. The results demonstrate the effect of shielding against the high inactivation potential of extraterrestrial solar UV radiation, which limits the chances of survival of even the highly UV-resistant strain of B. subtilis MW01 in the harsh environments of outer space and the martian surface.     

UV photochemistry of DNA in vitro and in Bacillus subtilis spores at earth-ambient and low atmospheric pressure: implications for spore survival on other planets or moons in the solar system

Two major parameters influencing the survival of Bacillus subtilis spores in space and on bodies within the Solar System are UV radiation and vacuum, both of which induce inactivating damage to DNA. To date, however, spore survival and DNA photochemistry have been explored only at the extremes of Earth-normal atmospheric pressure (101.3 kPa) and at simulated space vacuum (10(-3)-10(-6) Pa). In this study, wild-type spores, mutant spores lacking alpha/beta-type small, acid-soluble spore proteins (SASP), naked DNA, and complexes between SASP SspC and DNA were exposed simultaneously to UV (254 nm) at intermediate pressure (1-2 Pa), and the UV photoproducts cis,syn-thymine-thymine cyclobutane dimer (c,sTT), trans,syn-thymine-thymine cyclobutane dimer (t,sTT), and "spore photoproduct" (SP) were quantified. At 101.3 kPa, UV-treated wild-type spores accumulated only SP, but spores treated with UV radiation at 1-2 Pa exhibited a spectrum of DNA damage similar to that of spores treated at 10(-6) Pa, with accumulation of SP, c,sTT, and t,sTT. The presence or absence of alpha/beta-type SASP in spores was partly responsible for the shift observed between levels of SP and c,sTT, but not t,sTT. The changes observed in spore DNA photochemistry at 1-2 Pa in vivo were not reproduced by irradiation of naked DNA or SspC:DNA complexes in vitro, suggesting that factors other than SASP are involved in spore DNA photochemistry at low pressure.     

Resistance of bacterial endospores to outer space for planetary protection purposes--experiment PROTECT of the EXPOSE-E mission

Spore-forming bacteria are of particular concern in the context of planetary protection because their tough endospores may withstand certain sterilization procedures as well as the harsh environments of outer space or planetary surfaces. To test their hardiness on a hypothetical mission to Mars, spores of Bacillus subtilis 168 and Bacillus pumilus SAFR-032 were exposed for 1.5 years to selected parameters of space in the experiment PROTECT during the EXPOSE-E mission on board the International Space Station. Mounted as dry layers on spacecraft-qualified aluminum coupons, the "trip to Mars" spores experienced space vacuum, cosmic and extraterrestrial solar radiation, and temperature fluctuations, whereas the "stay on Mars" spores were subjected to a simulated martian environment that included atmospheric pressure and composition, and UV and cosmic radiation. The survival of spores from both assays was determined after retrieval. It was clearly shown that solar extraterrestrial UV radiation (λ≥110?nm) as well as the martian UV spectrum (λ≥200?nm) was the most deleterious factor applied; in some samples only a few survivors were recovered from spores exposed in monolayers. Spores in multilayers survived better by several orders of magnitude. All other environmental parameters encountered by the "trip to Mars" or "stay on Mars" spores did little harm to the spores, which showed about 50% survival or more. The data demonstrate the high chance of survival of spores on a Mars mission, if protected against solar irradiation. These results will have implications for planetary protection considerations.     

Survival and germinability of Bacillus subtilis spores exposed to simulated Mars solar radiation: implications for life detection and planetary protection

Bacterial spores have been considered as microbial life that could survive interplanetary transport by natural impact processes or human spaceflight activity. Deposition of terrestrial microbes or their biosignature molecules onto the surface of Mars could negatively impact life detection experiments and planetary protection measures. Simulated Mars solar radiation, particularly the ultraviolet component, has been shown to reduce spore viability, but its effect on spore germination and resulting production of biosignature molecules has not been explored. We examined the survival and germinability of Bacillus subtilis spores exposed to simulated martian conditions that include solar radiation. Spores of B. subtilis that contain luciferase resulting from expression of an sspB-luxAB gene fusion were deposited on aluminum coupons to simulate deposition on spacecraft surfaces and exposed to simulated Mars atmosphere and solar radiation. The equivalent of 42 min of simulated Mars solar radiation exposure reduced spore viability by nearly 3 logs, while germination-induced bioluminescence, a measure of germination metabolism, was reduced by less than 1 log. The data indicate that spores can retain the potential to initiate germination-associated metabolic processes and produce biological signature molecules after being rendered nonviable by exposure to Mars solar radiation.     

Survival of Bacillus pumilus spores for a prolonged period of time in real space conditions

To prevent forward contamination and maintain the scientific integrity of future life-detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated surfaces, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UV radiation and peroxide treatment. Subsequently, B. pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions via the European Technology Exposure Facility (EuTEF). After 18 months of exposure in the EXPOSE facility of the European Space Agency (ESA) on EuTEF under dark space conditions, SAFR-032 spores showed 10-40% survivability, whereas a survival rate of 85-100% was observed when these spores were kept aboard the ISS under dark simulated martian atmospheric conditions. In contrast, when UV (>110?nm) was applied on SAFR-032 spores for the same time period and under the same conditions used in EXPOSE, a ～7-log reduction in viability was observed. A parallel experiment was conducted on Earth with identical samples under simulated space conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (～3-log reduction in viability for "UV-Mars," and ～4-log reduction in viability for "UV-Space"). A comparative proteomics analysis indicated that proteins conferring resistant traits (superoxide dismutase) were present in higher concentration in space-exposed spores when compared to controls. Also, the first-generation cells and spores derived from space-exposed samples exhibited elevated UVC resistance when compared with their ground control counterparts. The data generated are important for calculating the probability and mechanisms of microbial survival in space conditions and assessing microbial contaminants as risks for forward contamination and in situ life detection.     

Surface characteristics of spacecraft components affect the aggregation of microorganisms and may lead to different survival rates of bacteria on Mars landers

Layers of dormant endospores of Bacillus subtilis HA101 were applied to eight different spacecraft materials and exposed to martian conditions of low pressure (8.5 mbar), low temperature (-10 degrees C), and high CO(2) gas composition and irradiated with a Mars-normal ultraviolet (UV-visible- near-infrared spectrum. Bacterial layers were exposed to either 1 min or 1 h of Mars-normal UV irradiation, which simulated clear-sky conditions on equatorial Mars (0.1 tau). When exposed to 1 min of Mars UV irradiation, the numbers of viable endospores of B. subtilis were reduced three to four orders of magnitude for two brands of aluminum (Al), stainless steel, chemfilm-treated Al, clear-anodized Al, and black-anodized Al coupons. In contrast, bacterial survival was reduced only one to two orders of magnitude for endospores on the non-metal materials astroquartz and graphite composite when bacterial endospores were exposed to 1 min of Mars UV irradiation. When bacterial monolayers were exposed to 1 h of Mars UV irradiation, no viable bacteria were recovered from the six metal coupons listed above. In contrast, bacterial survival was reduced only two to three orders of magnitude for spore layers on astroquartz and graphite composite exposed to 1 h of Mars UV irradiation. Scanning electron microscopy images of the bacterial monolayers on all eight spacecraft materials revealed that endospores of B. subtilis formed large aggregates of multilayered spores on astroquartz and graphite composite, but not on the other six spacecraft materials. It is likely that the formation of multilayered aggregates of endospores on astroquartz and graphite composite is responsible for the enhanced survival of bacterial cells on these materials.     

Mutagenesis in bacterial spores exposed to space and simulated martian conditions: data from the EXPOSE-E spaceflight experiment PROTECT

As part of the PROTECT experiment of the EXPOSE-E mission on board the International Space Station (ISS), the mutagenic efficiency of space was studied in spores of Bacillus subtilis 168. After 1.5 years' exposure to selected parameters of outer space or simulated martian conditions, the rates of induced mutations to rifampicin resistance (Rif(R)) and sporulation deficiency (Spo(-)) were quantified. In all flight samples, both mutations, Rif(R) and Spo(-), were induced and their rates increased by several orders of magnitude. Extraterrestrial solar UV radiation (>110?nm) as well as simulated martian UV radiation (>200?nm) led to the most pronounced increase (up to nearly 4 orders of magnitude); however, mutations were also induced in flight samples shielded from insolation, which were exposed to the same conditions except solar irradiation. Nucleotide sequencing located the Rif(R) mutations in the rpoB gene encoding the β-subunit of RNA polymerase. Mutations isolated from flight and parallel mission ground reference (MGR) samples were exclusively localized to Cluster I. The 21 Rif(R) mutations isolated from the flight experiment showed all a C to T transition and were all localized to one hotspot: H482Y. In mutants isolated from the MGR, the spectrum was wider with predicted amino acid changes at residues Q469K/L/R, H482D/P/R/Y, and S487L. The data show the unique mutagenic power of space and martian surface conditions as a consequence of DNA injuries induced by solar UV radiation and space vacuum or the low pressure of Mars.     

Project Icarus: A review of local interstellar medium properties of relevance for space missions to the nearest stars

I review those properties of the interstellar medium within 15 light-years of the Sun, which will be relevant for the planning of future rapid (v≥0.1c) interstellar space missions to the nearest stars. As the detailed properties of the local interstellar medium (LISM) may only become apparent after interstellar probes have been able to make in situ measurements, the first such probes will have to be designed conservatively with respect to what can be learned about the LISM from the immediate environment of the Solar System. It follows that studies of interstellar vehicles should assume the lowest plausible density when considering braking devices, which rely on transferring momentum from the vehicle to the surrounding medium, but the highest plausible densities when considering possible damage caused by the impact of the vehicle with interstellar material. Some suggestions for working values of these parameters are provided. This paper is a submission of the Project Icarus Study Group.     

Microcolonial fungi: survival potential of terrestrial vegetative structures

So far mainly spores or other "differentiated-for-survival" structures were considered to be resistant against extreme environmental constraints (including extraterrestrial challenges). Microcolonial fungi (MCF) are unique growth structures formed by eukaryotic microorganisms inhabiting rock varnish surfaces in terrestrial deserts. They are here proposed as a new object for exobiological study. Sun-exposed desert rocks provide surface habitats with intense solar radiation, a scarce water supply, drastic changes in temperature, and episodic to sporadic availability of nutrients. These challenging conditions reduce the diversity of life to MCF, whose resistance to desiccation and tolerance for ultraviolet (UV) radiation make them survival specialists. Based upon our studies of MCF, we propose that the following mechanisms are universally employed for survival on rock surfaces: (1) compact tissue-like colony organization formed by thermodynamically optimal round cells embedded in extracellular polymeric substances, (2) the presence of several types of UV-absorbing compounds (melanins and mycosporines) and antioxidants (carotenoids, melanins, and mycosporines) that convey multiple stress resistance to desiccation, temperature, and irradiation changes, and (3) intracellular developmental mechanisms typical for these structures.     

Microbial survival rates of Escherichia coli and Deinococcus radiodurans under low temperature, low pressure, and UV-Irradiation conditions, and their relevance to possible Martian life

Viability rates were determined for microbial populations of Escherichia coli and Deinococcus radiodurans under the environmental stresses of low temperature (-35 degrees C), low-pressure conditions (83.3 kPa), and ultraviolet (UV) irradiation (37 W/m(2)). During the stress tests the organisms were suspended in saltwater soil and freshwater soil media, at variable burial depths, and in seawater. Microbial populations of both organisms were most susceptible to dehydration stress associated with low-pressure conditions, and to UV irradiation. However, suspension in a liquid water medium and burial at larger depths (5 cm) improved survival rates markedly. Our results indicate that planetary surfaces that possess little to no atmosphere and have low water availability do not constitute a favorable environment for terrestrial microorganisms.     

掺铈玻璃型二次表面镜热控性能稳定性研究

对卫星用掺铈玻璃型二次表面镜(SSM)的热控性能在空间辐照环境中的长期稳定性进行了研究。给出了经紫外、电子和质子模拟辐照试验后，导电和非导电掺传玻璃型二次表面镜热控性能的数据，并对其衰退机理进行了分析。结果表明，掺铈玻璃型二次表面镜的热控性能在地球同步轨道空间辐照环境中的长期稳定性可以满足卫星设计要求。     

A comment on "the far future of exoplanet direct characterization"--the case for interstellar space probes

Following on from ideas presented in a recent paper by Schneider et al. on "The Far Future of Exoplanet Direct Characterization," I argue that they have exaggerated the technical obstacles to performing such "direct characterization" by means of fast (order 0.1c) interstellar space probes. A brief summary of rapid interstellar spaceflight concepts that may be found in the literature is presented. I argue that the presence of interstellar dust grains, while certainly something that will need to be allowed for in interstellar vehicle design, is unlikely to be the kind of showstopper suggested by Schneider et al. Astrobiology as a discipline would be a major beneficiary of developing an interstellar spaceflight capability, albeit in the longer term, and I argue that astrobiologists should keep an open mind to the possibilities.     

含磷聚酰亚胺薄膜在原子氧环境中的降解研究

文章考察了自行合成的含磷聚酰亚胺（PI）薄膜在模拟原子氧环境中的降解行为。扫描电子显微镜（SEM）、X射线光电子能谱（XPS）等测试结果表明:在原子氧辐照过程中,含磷PI薄膜表面的磷元素与氧元素含量增加,原子结合能也增大,意味着在PI表面形成了含磷钝化层。该钝化层进一步阻止了PI次表面层被侵蚀,使含磷PI薄膜表现出了抗原子氧侵蚀能力,其在模拟原子氧环境中的质量损失率远低于Kapton薄膜。     

太阳模拟器复杂构型卫星稳态 温度场的数值模拟

文章基于蒙特卡罗-区域分解法,建立太阳模拟器内复杂几何构型卫星稳态温度场数学模型;利用数值模拟求解卫星表面太阳辐照面积、红外和太阳辐射传递系数,得到不同辐照条件下卫星的温度分布;分析太阳辐照的非均匀性和不稳定性对温度分布的影响。计算结果表明:蒙特卡罗-区域分解法对于太阳模拟器模拟空间外热流的仿真研究是一种非常有效的方法;太阳辐照非均匀性对温度分布的影响大于不稳定性所产生的影响。     

A narratological approach to interpreting and designing interstellar messages

Contemporary narratology (narrative theory) offers a useful framework for interpreting interstellar messages that have already been sent to potential extraterrestrial recipients, as well as for designing messages that may be transmitted in the future. In this paper, narratological concepts are used to analyze in depth a single interstellar message sequence, elucidating methods by which various parts of speech (nouns, verbs, adjectives, and adverbs) can be paired with pictures to describe the human body in motion. The concept of focalization is applied to the message sequence's use of isolation and magnification, which highlight the structure and function of the human body and its constituent parts. The challenges of interpreting gaps within narratives, as well as the setting in which events occur, are considered. The importance of closure in providing a fitting end to narratives is examined, and the plausibility of creating images that could be interpreted correctly by extraterrestrial intelligence is assessed. Narratological concepts examined here, as well as additional aspects of narrative, provide important resources for future work in interpreting and designing interstellar messages.     

A search for optical beacons: implications of null results

Over the past few years a series of searches for interstellar radio beacons have taken place using the Parkes radio telescope. Here we report hitherto unpublished results from a search for optical beacons from 60 solar-type stars using the Perth-Lowell telescope. We discuss the significance of the null results from these searches, all of which were based on the interstellar contact channel hypothesis. While the null results of all searches to date can be explained simply by the nonexistence of electromagnetically communicating life elsewhere in the Milky Way, four other possible explanations that do not preclude its existence are proposed: (1) Extraterrestrial civilizations desiring to make contact through the use of electromagnetic beacons have a very low density in the Milky Way. (2) The interstellar contact channel hypothesis is incorrect, and beacons exist at frequencies that have not yet been searched. (3) The search has been incomplete in terms of sensitivity and/or target directions: Beacons exist, but more sensitive equipment and/or more searching is needed to achieve success. (4) The search has occurred before beacon signals can be expected to have arrived at the Earth, and beacon signals may be expected in the future. Based on consideration of the technology required for extraterrestrial civilizations to identify target planets, we argue that the fourth possibility is likely to be valid and that powerful, easily detectable beacons could be received in coming centuries.     

Radiation hazard of relativistic interstellar flight

From the point of view of radiation safety, interstellar space is not an empty void. Interstellar gas and cosmic rays, which consist of hydrogen and helium nucleons, present a severe radiation hazard to crew and electronics aboard a relativistic interstellar ship. Of the two, the oncoming relativistic flow of interstellar gas produces the most intense radiation. A protection shield will be needed to block relativistic interstellar gas that can also absorb most of the cosmic rays which, as a result of relativistic aberration, forms into a beamed flow propagating toward the front of the spaceship.     

Is interstellar archeology possible?

Searching for signatures of cosmic-scale archeological artifacts such as Dyson spheres is an interesting alternative to conventional radio SETI. Uncovering such an artifact does not require the intentional transmission of a signal on the part of the original civilization. This type of search is called interstellar archeology or sometimes cosmic archeology. A variety of interstellar archeology signatures is discussed including non-natural planetary atmospheric constituents, stellar doping, Dyson spheres, as well as signatures of stellar, and galactic-scale engineering. The concept of a Fermi bubble due to interstellar migration is reviewed in the discussion of galactic signatures. These potential interstellar archeological signatures are classified using the Kardashev scale. A modified Drake equation is introduced. With few exceptions interstellar archeological signatures are clouded and beyond current technological capabilities. However SETI for so-called cultural transmissions and planetary atmosphere signatures are within reach.