Critical issues in connection with human missions to Mars: protection of and from the Martian environment.

Human missions to Mars are planned to happen within this century. Activities associated therewith will interact with the environment of Mars in two reciprocal ways: (i) the mission needs to be protected from the natural environmental elements that can be harmful to human health, the equipment or to their operations; (ii) the specific natural environment of Mars should be protected so that it retains its value for scientific and other purposes. The following environmental elements need to be considered in order to protect humans and the equipment on the planetary surface: (i) cosmic ionizing radiation, (ii) solar particle events; (iii) solar ultraviolet radiation; (iv) reduced gravity; (v) thin atmosphere; (vi) extremes in temperatures and their fluctuations; and (vii) surface dust. In order to protect the planetary environment, the requirements for planetary protection as adopted by COSPAR for lander missions need to be revised in view of human presence on the planet. Landers carrying equipment for exobiological investigations require special consideration to reduce contamination by terrestrial microorganisms and organic matter to the greatest feasible extent. Records of human activities on the planet's surface should be maintained in sufficient detail that future scientific experimenters can determine whether environmental modifications have resulted from explorations.     

Planetary protection considerations for MarsNet and Mars sample return missions.

The ESA MarsNet mission proposal consists most probably of a trio of Mars landers. These landers each contain a variety of scientific equipment. The network of stations demands for a definition of its planetary protection requirements. With respect to the MarsNet mission only forward contamination problems will be considered. Future involvement of European efforts in planetary exploration including sample returns will also raise the problem of back contamination. A tradeoff study for the overall scientific benefit with respect to the approximative cost is necessary. Planetary protection guide-lines will be proposed by an interdisciplinary and international board of experts working in the fields of both biology and planetary science. These guide-lines will have to be flexible in order to be modified with respect to new research results, e.g. on adaptation of microorganisms to extreme (space) conditions. Experiments on the survival of microorganisms at conditions of simulated Mars surface and subsurface will have to be conducted in order to obtain a baseline data collection as a reference standard for future guide-lines.     

The conceptual design of a hybrid life support system based on the evaluation and comparison of terrestrial testbeds.

This report summarizes a trade study of different options of a bioregenerative Life Support System (LSS) and a subsequent conceptual design of a hybrid LSS. The evaluation was based mainly on the terrestrial testbed projects MELISSA (ESA) and BIOS (Russia). In addition, some methods suggested by the Advanced Life Support Project (NASA) were considered. Computer models, including mass flows were established for each of the systems with the goal of closing system loops to the extent possible. In order to cope with the differences in the supported crew size and provided nutrition, all systems were scaled for supporting a crew of six for a 780 day Mars mission (180 days transport to Mars; 600 days surface period) as given in the NASA Design Reference Mission Scenario [Hoffman, S.J., Kaplan, D.L. Human exploration of Mars: the Reference Mission of the NASA Mars Exploratory Study, 1997]. All models were scaled to provide the same daily allowances, as of calories, to the crew. Equivalent System Mass (ESM) analysis was used to compare the investigated system models against each other. Following the comparison of the terrestrial systems, the system specific subsystem options for Food Supply, Solid Waste Processing, Water Management and Atmosphere Revitalization were evaluated in a separate trade study. The best subsystem technologies from the trade study were integrated into an overall design solution based on mass flow relationships. The optimized LSS is mainly a bioregenerative system, complemented by a few physico-chemical elements, with a total ESM of 18,088 kg, which is about 4 times higher than that of a pure physico-chemical LSS, as designed in an earlier study.     

Embryogenesis and organogenesis of Carausius morosus under spaceflight conditions

The influence of cosmic radiation and/or microgravity on insect development was studied during the 7 day German Spacelab Mission D1. Eggs of Carausius morosus of five stages differing in sensitivity to radiation and in capacity to regeneration were allowed to continue their development in the BIORACK 22°C incubator, either at microgravity conditions or on the 1 g reference centrifuge. Using the Biostack concept - eggs in monolayers were sandwiched between visual track detectors - and the 1 g reference centrifuge, we were able to separate radiation effects from microgravity effects and also from combined effects of these two factors in space. After retrieval, hatching rates, growth kinetics and anomaly frequencies were determined in the different test samples. The early stages of development turned out to be highly sensitive to single hits of cosmic ray particles as well as to the temporary exposure to microgravity during their development. In some cases, the combined action of radiation and microgravity even amplified the effects exerted by the single parameters of space. Hits by single HZE particles caused early effects, such as body anomalies, as well as late effects, such as retarded growth after hatching. Microgravity exposure lead to a reduced hatching rate. A synergistic action of HZE particle hits and microgravity was established in the unexpectedly high frequency of anomal larvae. However, it cannot be excluded, that cosmic background radiation or low LET HZE particles are also causally involved in damage observed in the microgravity samples.     

The potential of the lichen symbiosis to cope with the extreme conditions of outer space II: germination capacity of lichen ascospores in response to simulated space conditions.

Complementary to the already well-studied microorganisms, lichens, symbiotic organisms of the mycobiont (fungi) and the photobiont (algae), were used as "model systems" in which to examine the ecological potential to resist to extreme environments of outer space. Ascospores (sexual propagules of the mycobiont) of the lichens Fulgensia bracteata, Xanthoria elegans and Xanthoria parietina were exposed to selected space-simulating conditions (up to 16 h of space vacuum at 10(-3) Pa and UV radiation at 160 nm < or = lambda < or = 400 nm), while embedded in the lichen fruiting bodies. After exposure, the ascospores were discharged and their viability was tested as germination capacity on different culture media including those containing Mars regolith simulant. It was found that (i) the germination rate on media containing Mars regolith simulant was as high as on other mineral-containing media, (ii) if enclosed in the ascocarps, the ascospores survived the vacuum exposure, the UV-irradiation as well as the combined treatment of vacuum and UV to a high degree. In general, 50 % or more viable spores were recovered, with ascospores of X. elegans showing the highest survival. It is suggested that ascospores inside the ascocarps are well protected by the anatomical structure, the gelatinous layer and the pigments (parietin and carotene) against the space parameters tested.     

Thymine photoproduct formation and inactivation of intact spores of Bacillus subtilis irradiated with short wavelength UV (200-300nm) at atmospheric pressure and in vacuo.

Vacuum exposure renders the survival of spores of Bacillus subtilis approximately five times more sensitive to ultraviolet light irradiation than exposure under atmospheric conditions. The photoproduct formation in spores irradiated under ultrahigh vacuum (UHV) conditions is compared to the photoproduct formation in spores irradiated at atmospheric pressure. Compared to irradiation at atmospheric pressure, where only the "spore photoproduct" 5-thyminyl-5,6-dihydrothymine (TDHT) can be detected, two additional photoproducts, known as the c,s and t,s isomers of thymine dimer (T<>T) are produced in vacuo. The spectral efficiencies for photoproduct formation in spores under atmospheric and vacuum conditions are compared. Since there is no increased formation of TDHT after irradiation in vacuum, TDHT cannot be made responsible for the observed vacuum effect. "Vacuum specific" photoproducts may cause a synergistic response of spores to the simultaneous action of ultraviolet light (UV) and UHV. Three different mechanisms are discussed for the enhanced sensitivity of B. subtilis spores to UV radiation in vacuum. The experiments described contribute valuable research information on the chance for survival of microorganisms in outer space.     

Biological responses to space: results of the experiment "Exobiological Unit" of ERA on EURECA I.

Spores of different strains of Bacillus subtilis and the Escherichia coli plasmid pUC19 were exposed to selected conditions of space (space vacuum and/or defined wavebands and intensities of solar ultraviolet radiation) in the experiment ER 161 "Exobiological Unit" of the Exobiology Radiation Assembly (ERA) on board of the European Retrievable Carrier (EURECA). After the approximately 11 months lasting mission, their responses were studied in terms of survival, mutagenesis in the his (B. subtilis) or lac locus (pUC19), induction of DNA strand breaks, efficiency of DNA repair systems, and the role of external protective agents. The data were compared with those of a simultaneously running ground control experiment. The survival of spores treated with the vacuum of space, however shielded against solar radiation, is substantially increased, if they are exposed in multilayers and/or in the presence of glucose as protective, whereas all spores in "artificial meteorites", i.e. embedded in clays or simulated Martian soil, are killed. Vacuum treatment leads to an increase of mutation frequency in spores, but not in plasmid DNA. Extraterrestrial solar ultraviolet radiation is mutagenic, induces strand breaks in the DNA and reduces survival substantially; however, even at the highest fluences, i.e. 3 x 10(8) J m-2, a small but significant fraction of spores survives the insolation. Action spectroscopy confirms results of previous space experiments of a synergistic action of space vacuum and solar UV radiation with DNA being the critical target.     

Long-term survival of bacterial spores in space.

On board of the NASA Long Duration Exposure Facility (LDEF), spores of Bacillus subtilis in monolayers (10(6)/sample) or multilayers (10(8)/sample) were exposed to the space environment for nearly six years and their survival was analyzed after retrieval. The response to space parameters, such as vacuum (10(-6) Pa), solar electromagnetic radiation up to the highly energetic vacuum-ultraviolet range (10(9) J/m2) and/or cosmic radiation (4.8 Gy), was studied and compared to the results of a simultaneously running ground control experiment. If shielded against solar ultraviolet (UV)-radiation, up to 80 % of spores in multilayers survive in space. Solar UV-radiation, being the most deleterious parameter of space, reduces survival by 4 orders of magnitude or more. However, up to 10(4) viable spores were still recovered, even in completely unprotected samples. Substances, such as glucose or buffer salts serve as chemical protectants. With this 6 year study in space, experimental data are provided to the discussion on the likelihood of "Panspermia".     

Survival of microorganisms in space: A review

Spores of $\text{Bacillus subtilis}$ were exposed to selected factors of space (vacuum, solar UV radiation, heavy ions of cosmic radiation), and their response was studied after recovery. These investigations were supplemented by ground-based studies under simulated space conditions. The vacuum of space did not inactivate the spores. However, vacuum-induced structural changes in the DNA, and probably in the proteins, caused a supersensitivity to solar UV radiation. This phenomenon is caused by the production of specific photoproducts in DNA and protein, which cannot be removed by normal cellular repair processes. In vegetative bacterial cells, exposed to vacuum, cell dehydration led to damage of the cell membrane, which could be partly repaired during subsequent incubation. The high local effectiveness of the cosmic heavy ions further decreases the chance that spores can survive for any length of time in space. Nonetheless, a spore travelling through space and protected from ultraviolet radiation could possibly survive an interplanetary journey. Such a situation favors panspermia as a possible explanation for the origin of life.     

Inactivation, mutation induction and repair in Bacillus subtilis spores irradiated with heavy ions.

Studies on the response of bacterial spores to accelerated heavy ions (HZE particles) help in understanding problems of space radiobiology and exobiology. Layers of spores of Bacillus subtilis strains, differing in repair capabilities, were irradiated with accelerated boron, carbon and neon ions of linear energy transfer (LET) values up to 14000 MeV cm2/g. Inactivation as measured by loss of colony forming ability and induction of mutations as measured by reversion to histidine prototrophy and resistance to 150 micrograms/ml sodium azide were tested, as well as the influence of repair processes on these effects. For inactivation, the cross-sectional values sigma plotted as a function of LET follow a saturation curve. The plateau, which is reached around a LET of 2000 MeV cm2/g, occurs at 2.5 x 10(-9) cm2, a value in good agreement with the dimensions of the spore protoplast. Lethal damage produced at LET values < 2000 MeV cm2/g is reparable. Recombination repair is more effective than excision repair. At higher LET values, lethal damage could not be reconstituted by the repair mechanisms studied. In addition, at these high LET values, the frequency of induced mutations was drastically decreased. The data support the assumption of at least two qualitatively different types of lesion, depending on the LET of the affecting heavy ion.