Establishment of a rearing system of the extremotolerant tardigrade Ramazzottius varieornatus: a new model animal for astrobiology

Studies on the ability of multicellular organisms to tolerate specific environmental extremes are relatively rare compared to those of unicellular microorganisms in extreme environments. Tardigrades are extremotolerant animals that can enter an ametabolic dry state called anhydrobiosis and have high tolerance to a variety of extreme environmental conditions, particularly while in anhydrobiosis. Although tardigrades have been expected to be a potential model animal for astrobiological studies due to their excellent anhydrobiotic and extremotolerant abilities, few studies of tolerance with cultured tardigrades have been reported, possibly due to the absence of a model species that can be easily maintained under rearing conditions. We report the successful rearing of the herbivorous tardigrade, Ramazzottius varieornatus, by supplying the green alga Chlorella vulgaris as food. The life span was 35 +/- 16.4 d, deposited eggs required 5.7 +/- 1.1 d to hatch, and animals began to deposit eggs 9 d after hatching. The reared individuals of this species had an anhydrobiotic capacity throughout their life cycle in egg, juvenile, and adult stages. Furthermore, the reared adults in an anhydrobiotic state were tolerant of temperatures of 90 degrees C and -196 degrees C, and exposure to 99.8% acetonitrile or irradiation with 4000 Gy (4)He ions. Based on their life history traits and tolerance to extreme stresses, R. varieornatus may be a suitable model for astrobiological studies of multicellular organisms.     

Tardigrades as a potential model organism in space research

Exposure of living organisms to open space requires a high level of tolerance to desiccation, cold, and radiation. Among animals, only anhydrobiotic species can fulfill these requirements. The invertebrate phylum Tardigrada includes many anhydrobiotic species, which are adapted to survive in very dry or cold environmental conditions. As a likely by-product of the adaptations for desiccation and freezing, tardigrades also show a very high tolerance to a number of other, unnatural conditions, including exposure to ionizing radiation. This makes tardigrades an interesting candidate for experimental exposure to open space. This paper reviews the tolerances that make tardigrades suitable for astrobiological studies and the reported radiation tolerance in other anhydrobiotic animals. Several studies have shown that tardigrades can survive gamma-irradiation well above 1 kilogray, and desiccated and hydrated (active) tardigrades respond similarly to irradiation. Thus, tolerance is not restricted to the dry anhydrobiotic state, and I discuss the possible involvement of an efficient, but yet undocumented, mechanism for DNA repair. Other anhydrobiotic animals (Artemia, Polypedium), when dessicated, show a higher tolerance to gamma-irradiation than hydrated animals, possibly due to the presence of high levels of the protective disaccharide trehalose in the dry state. Tardigrades and other anhydrobiotic animals provide a unique opportunity to study the effects of space exposure on metabolically inactive but vital metazoans.     

Implications of subzero metabolic activity on long-term microbial survival in terrestrial and extraterrestrial permafrost

The survival of microorganisms over extended time frames in frozen subsurface environments may be limited by chemical (i.e., via hydrolysis and oxidation) and ionizing radiation-induced damage to chromosomal DNA. In an effort to improve estimates for the survival of bacteria in icy terrestrial and extraterrestrial environments, we determined rates of macromolecular synthesis at temperatures down to -15°C in bacteria isolated from Siberian permafrost (Psychrobacter cryohalolentis K5 and P. arcticus 273-4) and the sensitivity of P. cryohalolentis to ionizing radiation. Based on experiments conducted over ≈400 days at -15°C, the rates of protein and DNA synthesis in P. cryohalolentis were <1 to 16 proteins cell(-1) d(-1) and 83 to 150 base pairs (bp) cell(-1) d(-1), respectively; P. arcticus synthesized DNA at rates of 20 to 1625?bp cell(-1) d(-1) at -15°C under the conditions tested. The dose of ionizing radiation at which 37% of the cells survive (D(37)) of frozen suspensions of P. cryohalolentis was 136?Gy, which was ～2-fold higher (71?Gy) than identical samples exposed as liquid suspensions. Laboratory measurements of [(3)H]thymidine incorporation demonstrate the physiological potential for DNA metabolism at -15°C and suggest a sufficient activity is possible to offset chromosomal damage incurred in near-subsurface terrestrial and martian permafrost. Thus, our data imply that the longevity of microorganisms actively metabolizing within permafrost environments is not constrained by chromosomal DNA damage resulting from ionizing radiation or entropic degradation over geological time.     

Time profile of cosmic radiation exposure during the EXPOSE-E mission: the R3DE instrument

The aim of this paper is to present the time profile of cosmic radiation exposure obtained by the Radiation Risk Radiometer-Dosimeter during the EXPOSE-E mission in the European Technology Exposure Facility on the International Space Station's Columbus module. Another aim is to make the obtained results available to other EXPOSE-E teams for use in their data analysis. Radiation Risk Radiometer-Dosimeter is a low-mass and small-dimension automatic device that measures solar radiation in four channels and cosmic ionizing radiation as well. The main results of the present study include the following: (1) three different radiation sources were detected and quantified-galactic cosmic rays (GCR), energetic protons from the South Atlantic Anomaly (SAA) region of the inner radiation belt, and energetic electrons from the outer radiation belt (ORB); (2) the highest daily averaged absorbed dose rate of 426 μGy d(-1) came from SAA protons; (3) GCR delivered a much smaller daily absorbed dose rate of 91.1 μGy d(-1), and the ORB source delivered only 8.6 μGy d(-1). The analysis of the UV and temperature data is a subject of another article (Schuster et al., 2012 ).     

Effects of sublethal irradiation with helium ions (300 MeV/nucleon) on basic hematological parameters of mice

The aim of the experiment was to obtain new knowledge on the biological effectiveness of high-energy (300 MeV/nucleon) helium ions, which represent a part of the spectrum of cosmic rays. Male (CBA x C57BL)F1 mice, 4 months old, were exposed to a dose of 4 Gy helium ions (exposure rate 0.05 Gy/min). As a comparative standard irradiation the same dose of 4 Gy of 137Cs gamma-rays (exposure rate 0.07 Gy/min) was used. Material sampling was performed 6-8 h, 4 days and 9 days after irradiation for both experimental groups mentioned above. There were 7 animals in each group including the control group of non-irradiated mice. Eight basic hematological parameters of peripheral blood, bone marrow, spleen and thymus were studied. On day 4 after the irradiation with helium ions, the values of leukocyte counts in peripheral blood, bone marrow cellularity and spleen cellularity were reduced to about 10% of the respective control values while the decline after irradiation with gamma-rays amounted to about 50%. These and other results presented reflect a high relative biological effectiveness of 300 MeV/nucleon helium ions.     

大容量Flash存储器空间辐射效应试验研究

分析了商用Flash存储器应用于航天器时应考虑的空间辐射效应和机理,并利用钴-60γ射线和重离子加速器对韩国三星公司生产的大容量Flash存储器K9XXG08UXA系列进行了抗电离总剂量试验和抗单粒子试验,以评估其空间应用可行性。试验结果显示：这一系列存储器的累积电离总剂量为50krad（Si）时,器件部分数据丢失,重...     

Comparative survival analysis of Deinococcus radiodurans and the haloarchaea Natrialba magadii and Haloferax volcanii exposed to vacuum ultraviolet irradiation

The haloarchaea Natrialba magadii and Haloferax volcanii, as well as the radiation-resistant bacterium Deinococcus radiodurans, were exposed to vacuum UV (VUV) radiation at the Brazilian Synchrotron Light Laboratory. Cell monolayers (containing 10(5) to 10(6) cells per sample) were prepared over polycarbonate filters and irradiated under high vacuum (10(-5) Pa) with polychromatic synchrotron radiation. N. magadii was remarkably resistant to high vacuum with a survival fraction of (3.77±0.76)×10(-2), which was larger than that of D. radiodurans (1.13±0.23)×10(-2). The survival fraction of the haloarchaea H. volcanii, of (3.60±1.80)×10(-4), was much smaller. Radiation resistance profiles were similar between the haloarchaea and D. radiodurans for fluences up to 150?J m(-2). For fluences larger than 150?J m(-2), there was a significant decrease in the survival of haloarchaea, and in particular H. volcanii did not survive. Survival for D. radiodurans was 1% after exposure to the higher VUV fluence (1350?J m(-2)), while N. magadii had a survival lower than 0.1%. Such survival fractions are discussed regarding the possibility of interplanetary transfer of viable microorganisms and the possible existence of microbial life in extraterrestrial salty environments such as the planet Mars and Jupiter's moon Europa. This is the first work to report survival of haloarchaea under simulated interplanetary conditions.     

Response of Phaseolus vulgaris L. plants to low-let ionizing radiation: Growth and oxidative stress

The scenarios for the long-term habitation of space platforms and planetary stations involve plants as fundamental part of Bioregenerative Life Support Systems (BLSS) to support the crew needs. Several constraints may limit plant growth in space: among them ionizing radiation is recognized to severely affect plant cell at morphological, physiological and biochemical level. In this work, plants of Phaseolus vulgaris L. were subjected to four different doses of X-rays (0.3, 10, 50 and 100 Gy) in order to assess the effects of ionizing radiation on this species and to analyze possible mechanisms carried out to overcome the radiation injuries. The effects of X-rays on plant growth were assessed by measuring stem elongation, number of internodes and leaf dry weight. The integrity of photosynthetic apparatus was evaluated by photosynthetic pigment composition and ribulose 1,5-bisphosphate carboxylase (Rubisco) activity, whereas changes in total antioxidant pool and glutathione S transferase activity (GST) were utilized as markers of oxidative stress. The distribution of phenolic compounds in leaf tissues as natural shielding against radiation was also determined.Irradiation of plants at 0.3 and 10 Gy did not determine differences in all considered parameters as compared to control. On the contrary, at 50 and 100 Gy a reduction of plant growth and a decrease in photosynthetic pigment content, as well as an increase in phenolic compounds and a decrease in total antioxidant content and GST activity were found. Only a slight reduction of Rubisco activity in leaves irradiated at 50 and 100 Gy was found. The overall results indicate P. vulgaris as a species with a good potential to face ionizing radiation and suggest its suitability for utilization in BLSSs.     

Microbial diversity of Indian Ocean hydrothermal vent plumes: microbes tolerant of desiccation, peroxide exposure, and ultraviolet and gamma-irradiation

The microbial diversity of Kali chimney plumes, part of a hydrothermal vent field in the Rodriguez Triple Junction, Indian Ocean (depth approximately 2,240 m), was examined in an attempt to discover "extremotolerant" microorganisms that have evolved unique resistance capabilities to this harsh environment. Water and sediment samples were collected from the vent and from sediments located at various distances (2-20 m) away from and surrounding the chimney. Samples were screened for hypertolerant microbes that are able to withstand multiple stresses. A total of 46 isolates were selected for exposure to a number of perturbations, such as heat shock, desiccation, H(2)O(2), and ultraviolet (UV) and gamma-irradiation. The survival of Psychrobacter sp. L0S3S-03b following exposure to >1,000 J/m(2) UV(254) radiation was particularly intriguing amid a background of varying levels of resistance. Vegetative cells of this non-spore-forming microbe not only survived all of the treatments, but also exhibited a 90% lethal dose of 30 s when exposed to simulated martian UV radiation and a 100% lethal dose of 2 min when exposed to full spectrum UV, which is comparable to findings for bacterial endospores.     

Cosmic radiation exposure of biological test systems during the EXPOSE-E mission

In the frame of the EXPOSE-E mission on the Columbus external payload facility EuTEF on board the International Space Station, passive thermoluminescence dosimeters were applied to measure the radiation exposure of biological samples. The detectors were located either as stacks next to biological specimens to determine the depth dose distribution or beneath the sample carriers to determine the dose levels for maximum shielding. The maximum mission dose measured in the upper layer of the depth dose part of the experiment amounted to 238±10 mGy, which relates to an average dose rate of 408±16 μGy/d. In these stacks of about 8?mm height, the dose decreased by 5-12% with depth. The maximum dose measured beneath the sample carriers was 215±16 mGy, which amounts to an average dose rate of 368±27 μGy/d. These values are close to those assessed for the interior of the Columbus module and demonstrate the high shielding of the biological experiments within the EXPOSE-E facility. Besides the shielding by the EXPOSE-E hardware itself, additional shielding was experienced by the external structures adjacent to EXPOSE-E, such as EuTEF and Columbus. This led to a dose gradient over the entire exposure area, from 215±16 mGy for the lowest to 121±6 mGy for maximum shielding. Hence, the doses perceived by the biological samples inside EXPOSE-E varied by 70% (from lowest to highest dose). As a consequence of the high shielding, the biological samples were predominantly exposed to galactic cosmic heavy ions, while electrons and a significant fraction of protons of the radiation belts and solar wind did not reach the samples.     

Recent results of the joint ESA-DARA/IBMP experiments Biokosmos "Seeds"

Comparison of experimental data obtained from short (SDEF) and long duration exposure flights (LDEF) have recently led to results which will be significant for longer and/or repeated sojourn of man in space. Under orbital conditions biological stress and damage are induced in test subjects by cosmic radiation, especially the high energetic, densely ionizing component of heavy ions. Plant seeds were successful model systems for a biotest in studying the physiological damages and mutagenic effect caused by ionizing cosmic radiation in particular stem cells. Dosimetrically, the subdivision into charge- and Let-groups reveals the contribution of the intermediate group (LET = 350-1000 MeV/cm) due to the medium heavy ions (Z = 6-10). Their relative contribution increases with the lower inclination of the orbit of LDEF-1; on the other hand, the total fluence becomes higher with longer duration of the flight. The observed endpoints of the biological radiation damage hint at a correlation with particle dose rate rather than with the dose; additionally, data on shielding effects inside and outside the space craft and its exposure were gained from the different SDEF- and LDEF-missions.     

Assessing antibody microarrays for space missions: effect of long-term storage, gamma radiation, and temperature shifts on printed and fluorescently labeled antibodies

Antibody microarrays are becoming frequently used tools for analytical purposes. A key factor for optimal performance is the stability of the immobilized (capturing) antibodies as well as those that have been fluorescently labeled to achieve the immunological test (tracers). This is especially critical for long-distance transport, field testing, or planetary exploration. A number of different environmental stresses may affect the antibody integrity, such as dryness, sudden temperature shift cycles, or, as in the case of space science, exposure to large quantities of the highly penetrating gamma radiation. Here, we report on the effect of certain stabilizing solutions for long-term storage of printed antibody microarrays under different conditions. We tested the effect of gamma radiation on printed and freeze- or vacuum-dried fluorescent antibodies at working concentrations (tracer antibodies), as well as the effect of multiple cycles of sudden and prolonged temperature shifts on the stability of fluorescently labeled tracer antibody cocktails. Our results show that (i) antibody microarrays are stable at room temperature when printed on stabilizing spotting solutions for at least 6 months, (ii) lyophilized and vacuum-dried fluorescently labeled tracer antibodies are stable for more than 9 months of sudden temperature shift cycles (-20°C to 25°C and 50°C), and (iii) both printed and freeze- or vacuum-dried fluorescent tracer antibodies are stable after several-fold excess of the dose of gamma radiation expected during a mission to Mars. Although different antibodies may exhibit different susceptibilities, we conclude that, in general, antibodies are suitable for use in planetary exploration purposes if they are properly treated and stored with the use of stabilizing substances.     

Low-temperature ionizing radiation resistance of Deinococcus radiodurans and Antarctic Dry Valley bacteria

The high flux of cosmic rays onto the unshielded surface of Mars poses a significant hazard to the survival of martian microbial life. Here, we determined the survival responses of several bacterial strains to ionizing radiation exposure while frozen at a low temperature characteristic of the martian near-subsurface. Novel psychrotolerant bacterial strains were isolated from the Antarctic Dry Valleys, an environmental analogue of the martian surface, and identified by 16S rRNA gene phylogeny as representatives of Brevundimonas, Rhodococcus, and Pseudomonas genera. These isolates, in addition to the known radioresistant extremophile Deinococcus radiodurans, were exposed to gamma rays while frozen on dry ice (-79°C). We found D. radiodurans to exhibit far greater radiation resistance when irradiated at -79°C than was observed in similar studies performed at higher temperatures. This greater radiation resistance has important implications for the estimation of potential survival times of microorganisms near the martian surface. Furthermore, the most radiation resistant of these Dry Valley isolates, Brevundimonas sp. MV.7, was found to show 99% 16S rRNA gene similarity to contaminant bacteria discovered in clean rooms at both Kennedy and Johnson Space Centers and so is of prime concern to efforts in the planetary protection of Mars from our lander probes. Results from this experimental irradiation, combined with previous radiation modeling, indicate that Brevundimonas sp. MV.7 emplaced only 30?cm deep in martian dust could survive the cosmic radiation for up to 100,000 years before suffering 10? population reduction.     

轨道分子屏极高真空实验室设计技术研究

给出了一种新型的近地轨道极高真空分子屏实验室设计。作者计算了可变翼分子屏周围大气分子的入射及散射、分子屏材料放气在屏内的分子数流密度。通过调整分子屏的翼角,分子屏内的压力可以达到10-12Pa。计算结果指出:在近地轨道选择合适的分子屏参数,可以在极高真空环境下加工超纯材料。     

Using radiation risk for assessment of space radiation hazard

Radiation hazard caused by exposure during a spaceflight is characterized by radiobiological consequences at all levels of organism. These consequences have a stochastic nature. Even deterministic effects are basically random quantity having all attributes of such mathematical values. The radiation risk is defined in this case as an additional probability of health damage or as a death probability in extreme case. For the manned spaceflight additional peculiarity of a human’s exposure is added. A natural space radiation environment has a stochastic character because solar particle events and crew of a spacecraft can be exposed to dose from background level up to lethal one.The report presents a procedure of radiation risk assessment for quantitative expression of radiation hazard level during a flight and using this value for developing protection recommendations. It is emphasized that the risk value is connected specifically with the time interval of possible hazard’s existent. The form of risk representation must be chosen depending on a time scale of radiobiological processes induced by the exposure (expressing in fact the radiation hazard model). Surviving function specified for the crewmember mortality rate changed by the professional exposure must be used for risk calculation. Solar particle events determine a stochastic character of radiation environment in space that must be taken into account for a risk assessment. The reliability of radiation risk assessment can be used for this goal.     

Space radiation concerns for manned exploration.

Spaceflight exposes astronaut crews to natural ionizing radiation. To date, exposures in manned spaceflight have been well below the career limits recommended to NASA by the National Council of Radiation Protection and Measurements (NCRP). This will not be the case for long-duration exploratory class missions. Additionally. International Space Station (ISS) crews will receive higher doses than earlier flight crews. Uncertainties in our understanding of long-term bioeffects, as well as updated analyses of the Hiroshima. Nagasaki and Chernobyl tumorigenesis data, have prompted the NCRP to recommend further reductions by 30-50% for career dose limit guidelines. Intelligent spacecraft design and material selection can provide a shielding strategy capable of maintaining crew exposures within recommended guidelines. Current studies on newer radioprotectant compounds may find combinations of agents which further diminish the risk of radiation-induced bioeffects to the crew.     

Was Earth ever infected by martian biota? Clues from radioresistant bacteria

Here we propose that the radioresistance (tolerance to ionizing radiation) observed in several terrestrial bacteria has a martian origin. Multiple inconsistencies with the current view of radioresistance as an accidental side effect of tolerance to desiccation are discussed. Experiments carried out 25 years ago were reproduced to demonstrate that "ordinary" bacteria can develop high radioresistance ability after multiple cycles of exposure to high radiation dosages followed by cycles of recovery of the bacterial population. We argue that "natural" cycles of this kind could have taken place only on the martian surface, and we hypothesize that Mars microorganisms could have developed radioresistance in just several million years' time and, subsequently, have undergone transfer to Earth by way of martian meteorites. Our mechanism implies multiple and frequent exchanges of biota between Mars and Earth.     

A technique for estimating the probability of radiation-stimulated failures of integrated microcircuits in low-intensity radiation fields: Application to the Spektr-R spacecraft

In developing radio-electronic devices (RED) of spacecraft operating in the fields of ionizing radiation in space, one of the most important problems is the correct estimation of their radiation tolerance. The “weakest link” in the element base of onboard microelectronic devices under radiation effect is the integrated microcircuits (IMC), especially of large scale (LSI) and very large scale (VLSI) degree of integration. The main characteristic of IMC, which is taken into account when making decisions on using some particular type of IMC in the onboard RED, is the probability of non-failure operation (NFO) at the end of the spacecraft’s lifetime. It should be noted that, until now, the NFO has been calculated only from the reliability characteristics, disregarding the radiation effect. This paper presents the so-called “reliability” approach to determination of radiation tolerance of IMC, which allows one to estimate the probability of non-failure operation of various types of IMC with due account of radiation-stimulated dose failures. The described technique is applied to RED onboard the Spektr-R spacecraft to be launched in 2007.     

空间原子氧对低地球轨道航天器用粘结剂的腐蚀效应影响

原子氧是低地球轨道(leo)-种重要的空间环境因素,地面模拟试验是用于开展leo航天器备选材料原子氧环境评价的主要手段.利用北京卫星环境研究所的原子氧设备,对牌号为e-32的环氧树脂和牌号为gs414的硅橡胶两种粘结剂进行了评价试验.依据轨道参数与航天器两年的设计寿命,选定试验中原子氧的等效积分通量为1.4×1021atom/cm2.试验结果表明,环氧树脂样品出现了较大的质量损失,而硅橡胶样品的质量损失则相对较小.根据质量损失计算环氧树脂样品的原子氧反应率处于3.2×10-24～3.8×10-24cm3/atom之间.试验后的环氧树脂样品表面颜色变浅,硅橡胶样品的表面则呈现了玻璃化.对两种材料都进行了表面形貌分析,试验前后测试结果的对比分析表明原子氧的氧化作用使材料表面发生了较大的变化. abstract： atomic oxygen (ao) is prevalent in low earth orbit (leo). ground-based testing on ao exposure was performed to investigate the effects of atomic oxygen on materials to be used in leo spacecraft. two types of adhesive materials, e-32 epoxy and gs414 silicone, were tested in the atomic oxygen testing facility at beijing institute of spacecraft environment engineering (bisee). the equivalent atomic oxygen fluence in test was approximately 1.4×1021 atom/cm2 as determined based on two years' exposure on leo. significant mass losses of the epoxy adhesive samples were observed after the exposure to atomic oxygen, but relatively small mass changes were found in the silicone samples. the erosion yields of epoxy samples range from 3.2×10-24 cm3/atom to 3.8×10-24 cm3/atom. bleaching by atomic oxygen was found in the epoxy samples. the surface on the silicone sample was glossed after ao exposure. the external appearance of both kinds of materials was analyzed using scanning electron microscope (sem) and x-ray photoelectron spectroscopy (xps). a comparison between unexposed and exposed samples shows noticeable changes on the s     

环氧树脂648和TDE-85的质子辐照损伤效应研究

通过地面模拟空间环境质子辐照条件(质子能量Ep=150keV,束流密度A=2.0×1012cm-2·s-1,辐照剂量范围Φ=(0～5.0)×1016cm-2;真空度10-6Pa,环境温度120 K),研究了不同辐照剂量下环氧树脂浇注体648和TDE-85的质量损失、弯曲强度和表面相糙度的变化.试验结果表明,随着辐照剂量的增加,质量损失呈现先加速递增后趋于平缓的趋势;648的弯曲强度呈单边下降趋势,TDE-85的弯曲强度呈现先上升后下降的趋势,树脂表面产生了碳化效应,表面粗糙度发生了不同程度的变化.