Atmospheric dynamics and bioregenerative technologies in a soil-based ecological life support system: initial results from Biosphere 2.

Biosphere 2 is the first man-made, soil-based, bioregenerative life support system to be developed and tested. The utilization and amendment of local space resources, e.g. martian soil or lunar regolith, for agricultural and other purposes will be necessary if we are to minimize the requirement for Earth materials in the creation of long-term off-planet bases and habitations. Several of the roles soil plays in Biosphere 2 are 1) for air purification 2) as a key component in created wetland systems to recycle human and animal wastes and 3) as nutrient base for a sustainable agricultural cropping program. Initial results from the Biosphere 2 closure experiment are presented. These include the accelerated cycling rates due to small reservoir sizes, strong diurnal and seasonal fluxes in atmospheric CO2, an unexpected and continuing decline in atmospheric oxygen, overall maintenance of low levels of trace gases, recycling of waste waters through biological regeneration systems, and operation of an agriculture designed to provide diverse and nutritionally adequate diets for the crew members.     

Interaction of physical-chemical and biological regeneration processes in ecological Life Support Systems.

Catalytic combustion of inedible biomass of plants in ecological Life Support Systems (LSS) gives rise to gaseous oxides (CO2, NO2, SO2, etc.). Some of them are toxic for plants suppressing their photosynthesis and productivity. Experiments with "Bios-3" experimental LSS demonstrate that a decrease of photosynthetic productivity in a system with straw incineration can jeopardize its steady operation. Analysis of the situation by a mathematical model taking into account absorption parameters of the system in terms of toxic elements makes it possible to formulate requirements for the structure and operation of LSS to provide for its stability. Avenues for further investigation of the problem of toxic stability of LSS are proposed.     

Self-restoration of biocomponents as a mean to enhance Biological Life Support Systems reliability.

One of the key problems of long-term space missions is limited service life of units. The only exceptions are biological components of biological Life Support Systems--higher plants or microorganisms. These components are capable of self-restoration: after complete disintegration, they can appear again from seeds or spores. The estimate of failure intensity of BLSS regeneration component includes: a number of self-sustained sections of the regeneration component; permissible boost (how many times can productivity of a component be increased); time required to repair (restore) a component; the crew existence time, when all LSS regeneration components fail; failure rate of one section of a regeneration component. Evaluations show that for hydrogen-oxidizing bacteria and micro-algae very high reliability is achieved even for one or two sections. In the case of higher plants (due to low rate of self-restoration) bio-regenerative module has to be divided into 10 self-sustained sections operating simultaneously. These measures can decrease the probability of catastrophe by a factor of 10(6).     

Selection of candidate salad vegetables for controlled ecological life support system

Higher plants, as one of the essential biological components of CELSS, can supply food, oxygen and water for human crews during future long-duration space missions and Lunar/Mars habitats. In order to select suitable leaf vegetable varieties for our CELSS Experimental Facility (CEF), five varieties of lettuce (“Nenlvnaiyou”, “Dasusheng”, “Naichoutai”, “Dongfangkaixuan” and “Siji”), two of spinach (“Daye” and “Quanneng”), one of rape (“Jingyou No. 1”) and one of common sowthistle were grown and compared on the basis of edible biomass, and nutrient content. In addition, two series of experiments were conducted to study single leaf photosynthetic rates and transpiration rates at 30 days after planting, one which used various concentrations of CO₂ (500, 1000, 1500 and 2000 μmol mol⁻¹) and another which used various light intensities (100, 300, 500 and 700 μmol m⁻² s⁻¹). Results showed that lettuce cvs. “Nenlvnaiyou”, “Siji” and “Dasusheng” produced higher yields of edible biomass; common sowthisle would be a good source of β-carotene for the diet. Based on the collective findings, we selected three varieties of lettuce (“Nenlvnaiyou”, “Dasusheng” and “Siji”) and one of common sowthistle as the candidate crops for further research in our CEF. In addition, elevated CO₂ concentration increased the rates of photosynthesis and transpiration, and elevated light intensity increased the rate of photosynthesis for these varieties. These results can be useful for determining optimal conditions for controlling CO₂ and water fluxes between the crops and the overall CELSS.     

Phosphorus as a potential guide in the search for extinct life on Mars.

In contrast to the search for extant organisms, the quest for fossil remains of life on Mars need not be guided by the presence of water and organic compounds on the present surface. An appropriate tracer might be the element phosphorus which is a common constituent of living systems. Utilizing terrestrial analogues, it should preferentially exist in the form of sedimentary calcium phosphate (phosphorites), which would have readily resisted changing conditions on Mars. Moreover, higher ratios of P/Th in phosphorites in comparison to calcium phosphates from magmatic rocks give us the possibility to distinguish them from inorganically formed phosphorus deposits at or close to the Martian surface. Identification of anomalous phosphorus enrichments by remote sensing or in situ analysis could be promising approaches for selecting areas preferentially composed of rocks with remains of extinct life.     

Utilization of sweet potatoes in controlled ecological life support systems (CELSS).

A number of studies have selected the sweet potato as a potentially important crop for CELSS. Most hydroponic studies of sweet potatoes have been short term (<80 days). Full term (90 to 150 days) studies of sweet potatoes in hydroponic systems were needed to understand the physiology of storage root enlargement and to evaluate sweet potato production potential for CELSS. Early and late maturing sweet potato varieties were crown in hydroponic systems of different types--static with periodic replacement, flowing with and without recirculation, aggregate, and non-aggregate. In a flowing system with recirculation designed at Tuskegee University using the nutrient film technique (NFT), storage root yields as high as 1790 g were produced with an edible growth rate of up to 66 g m-2 d-1 and a harvest index as high as 89% under greenhouse conditions. Preliminary experiments indicated high yields can be obtained in controlled environmental chambers. Significant cultivar differences were found in all systems studied. Nutritive composition of storage roots and foliage were similar to field-grown plants. The results indicate great potential for sweet potato in CELSS.     

Simulated cometary matter as a test for enantiomer separating chromatography for use on comet 46P/Wirtanen.

The Cometary Sampling and Composition Experiment on board of European Space Agency's cornerstone mission ROSETTA is designed to identify organic molecules in cometary matter in situ by a combined pyrolysis gas chromatographic and mass spectrometric technique. Its capillary columns coated with chiral stationary phases received considerable attention, because they are designed for separations of non-complex enantiomers to allow the determination of enantiomeric ratios of cometary chiral organic compounds and consequently to provide information about the origin of molecular parity violation in biomolecules. To get gas chromatographic access to organic compounds on the comet, where macromolecules and complex organic polymers of low volatility are expected to make up the main organic ingredients, the combination of two injection techniques will be applied. The pyrolysis technique performed by heating cometary samples stepwise to defined temperatures in specific ovens resulting in thermochemolysis reactions of polymers and a chemical derivatization technique, in which the reagent dimethylformamide dimethylacetal assists pyrolysis derivatization reactions in producing methyl esters of polar monomers. The combination of the reagent assisted pyrolysis gas chromatographic technique with enantiomer separating chromatography was tested with laboratory-produced simulated cometary matter.     

The MELISSA pilot plant facility as as integration test-bed for advanced life support systems.

The different advances in the Micro Ecological Life Support System Alternative project (MELISSA), fostered and coordinated by the European Space Agency, as well as in other associated technologies, are integrated and demonstrated in the MELISSA Pilot Plant laboratory. During the first period of operation, the definition of the different compartments at an individual basis has been achieved, and the complete facility is being re-designed to face a new period of integration of all these compartments. The final objective is to demonstrate the potentiality of biological systems such as MELISSA as life support systems. The facility will also serve as a test bed to study the robustness and stability of the continuous operation of a complex biological system. This includes testing of the associated instrumentation and control for a safe operation, characterization of the chemical and microbial safety of the system, as well as tracking the genetic stability of the microbial strains used. The new period is envisaged as a contribution to the further development of more complete biological life support systems for long-term manned missions, that should be better defined from the knowledge to be gained from this integration phase. This contribution summarizes the current status of the Pilot Plant and the planned steps for the new period.     

Microbiological characterization of a regenerative life support system.

A Variable Pressure Plant Growth Chamber (VPGC), at the Johnson Space Center's (JSC) ground-based Regenerative Life Support Systems (RLSS) test bed, was used to produce crops of soil-grown lettuce. The crops and chamber were analyzed for microbiological diversity during lettuce growth and after harvest. Bacterial counts for the rhizosphere, spent nutrient medium, heat exchanger condensate, and atmosphere were approximately 10(11) Colony Forming Units (CFU) g-1 10(5) CFU ml-1, 10(5) CFU ml-1, and 600 CFU m-3, respectively. Pseudomonas was the predominant bacterial genus. Numbers of fungi were about 10(5) CFU g-1 in the rhizosphere, 4-200 CFU ml-1 in the spent nutrient medium, 110 CFU ml-1 in the heat exchanger condensate, and 3 CFU m-3 in the atmosphere. Fusarium and Trichoderma were the predominant fungal genera.     

NASA's Biomass Production Chamber: a testbed for bioregenerative life support studies.

The Biomass Production Chamber (BPC) located at Kennedy Space Center, FL, USA provides a large (20 m2 area, 113 m3 vol.), closed environment for crop growth tests for NASA's Controlled Ecological Life Support System (CELSS) program. Since the summer of 1988, the chamber has operated on a near-continuous basis (over 1200 days) without any major failures (excluding temporary power losses). During this time, five crops of wheat (64-86 days each), three crops of soybean (90 to 97 days), five crops of lettuce (28-30 days), and four crops of potato (90 to 105 days were grown, producing 481 kg of dry plant biomass, 196 kg edible biomass, 540 kg of oxygen, 94,700 kg of condensed water, and fixing 739 kg of carbon dioxide. Results indicate that total biomass yields were close to expected values for the given light input, but edible biomass yields and harvest indices were slightly lower than expected. Stand photosynthesis, respiration, transpiration, and nutrient uptake rates were monitored throughout growth and development of the different crops, along with the build-up of ethylene and other volatile organic compounds in the atmosphere. Data were also gathered on system hardware maintenance and repair, as well as person-hours required for chamber operation. Future tests will include long-term crop production studies, tests in which nutrients from waste treatment systems will be used to grow new crops, and multi-species tests.     

Neoplastic cell transformation by high-LET radiation: molecular mechanisms.

Experimental data on molecular mechanisms are essential for understanding the bioeffects of radiation and for developing biophysical models, which can help in determining the shape of dose-response curves at very low doses, e.g., doses less than 1 cGy. Although it has been shown that ionizing radiation can cause neoplastic cell transformation directly, that high-LET heavy ions in general can be more effective than photons in transforming cells, and that the radiogenic cell transformation is a multi-step process [correction of processes], we know very little about the molecular nature of lesions important for cell transformation, the relationship between lethal and transformational damages, and the evolution of initial damages into final chromosomal aberrations which alter the growth control of cells. Using cultured mouse embryo cells (C3H10T1/2) as a model system, we have collected quantitative data on dose-response curves for heavy ions with various charges and energies. An analysis of these quantitative data suggested that two DNA breaks formed within 80 angstroms may cause cell transformation and that two DNA breaks formed within 20 angstroms may be lethal. Through studies with restriction enzymes which produce DNA damages at specific sites, we have found that DNA double strand breaks, including both blunt- and cohesive-ended breaks, can cause cell transformation in vitro. These results indicate that DNA double strand breaks can be important primary lesions for radiogenic cell transformation and that blunt-ended double strand breaks can form lethal as well as transformational damages due to misrepair or incomplete repair in the cell. The RBE-LET relationship is similar for HGPRT gene mutation, chromosomal deletion, and cell transformation, suggesting common lesions may be involved in these radiation effects. The high RBE of high-LET radiation for cell killing and neoplastic cell transformation is most likely related to its effectiveness in producing DNA double strand breaks in mammalian cells. At present the role of oncogenes in radiation cell transformation is unclear.     

Biosphere 2: a prototype project for a permanent and evolving life system for Mars base.

As part of the ground-based preparation for creating long-term life systems needed for space habitation and settlement, Space Biospheres Ventures (SBV) is undertaking the Biosphere 2 project near Oracle, Arizona. Biosphere 2, currently under construction, is scheduled to commence its operations in 1991 with a two-year closure period with a crew of eight people. Biosphere 2 is a facility which will be essentialy materially-closed to exchange with the outside environment. It is open to information and energy flow. Biosphere 2 is designed to achieve a complex life-support system by the integration of seven areas or "biomes"--rainforest, savannah, desert, marsh, ocean, intensive agriculture and human habitat. Unique bioregenerative technologies, such as soil bed reactors for air purification, aquatic waste processing systems, real-time analytic systems and complex computer monitoring and control systems are being developed for the Biosphere 2 project. Its operation should afford valuable insight into the functioning of complex life systems necessary for long-term habitation in space. It will serve as an experimental ground-based prototype and testbed for the stable, permanent life systems needed for human exploration of Mars.     

Search for life on Mars: evaluation of techniques.

An important question for exobiology is, did life evolve on Mars? To answer this question, experiments must be conducted on the martian surface. Given current mission constraints on mass, power, and volume, these experiments can only be performed using proposed analytical techniques such as: electron microscopy, X-ray fluorescence, X-ray diffraction, alpha-proton backscatter, gamma-ray spectrometry, differential thermal analysis, differential scanning calorimetry, pyrolysis gas chromatography, mass spectrometry, and specific element detectors. Using prepared test samples consisting of 1% organic matter (bovine serum albumin) in palagonite and a mixture of palagonite, clays, iron oxides, and evaporites, it was determined that a combination of X-ray diffraction and differential thermal analysis coupled with gas chromatography provides the best insight into the chemistry, mineralogy, and geological history of the samples.     

The use of mineral crystals as bio-markers in the search for life on Mars.

Photographs that depict presumed fluvial features on the martian surface have led geologists to hypothesize that water flowed across the early martian terrain. From this, it has been further hypothesized that the surface and atmospheric conditions on early Mars were similar to those on early Earth. Because the oldest fossil evidence of life on Earth dates back to this early period, at least 3.5 billion years ago, the possibility exists that the early Martian environment could have also been conducive to the origin of life. To investigate this possibility, universal signatures or bio-markers indicative of past (or present) biological activity must be identified for use in the search for life on Mars. Several potentially applicable biomarkers have been identified and include: organics (e.g., specific classes of lipids and hopanes), suites of specific inorganic and organic compounds, as well as the isotopic ratios of C, N, and S. Unfortunately, all of these bio-markers may be of biologic or abiotic origin; these origins are often difficult to distinguish. Thus, the discovery of any one of these compounds alone is not a bio-marker. Because minerals produced under biologic control have distinctive crystallographies, morphologies, and isotopic ratios that distinguishable from abiotically produced minerals with the same chemical composition, and are stable through geologic time, we propose the use of minerals resulting from biologically controlled mineralization processes as bio-markers.     

Exponential evolution: implications for intelligent extraterrestrial life.

Some measures of biologic complexity, including maximal levels of brain development, are exponential functions of time through intervals of 10(6) to 10(9) yrs. Biological interactions apparently stimulate evolution but physical conditions determine the time required to achieve a given level of complexity. Trends in brain evolution suggest that other organisms could attain human levels within approximately 10(7) yrs. The number (N) and longevity (L) terms in appropriate modifications of the Drake Equation, together with trends in the evolution of biological complexity on Earth, could provide rough estimates of the prevalence of life forms at specified levels of complexity within the Galaxy. If life occurs throughout the cosmos, exponential evolutionary processes imply that higher intelligence will soon (10(9) yrs) become more prevalent than it now is. Changes in the physical universe become less rapid as time increases from the Big Bang. Changes in biological complexity may be most rapid at such later times. This lends a unique and symmetrical importance to early and late universal times.     

Volatile metabolites of higher plant crops as a photosynthesizing life support system component under temperature stress at different light intensities.

The effect of elevated temperatures of 35 and 45 degrees C (at the intensities of photosynthetically active radiation 322, 690 and 1104 micromoles m-2 s-1) on the photosynthesis, respiration, and qualitative and quantitative composition of the volatiles emitted by wheat (Triticum aestuvi L., cultivar 232) crops was investigated in growth chambers. Identification and quantification of more than 20 volatile compounds (terpenoids--alpha-pinene, delta 3 carene, limonene, benzene, alpha- and trans-caryophyllene, alpha- and gamma-terpinene, their derivatives, aromatic hydrocarbons, etc.) were conducted by gas chromatograph/mass spectrometry. Under light intensity of 1104 micromoles m-2 s-1 heat resistance of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. The action of elevated temperatures brought about variations in the rate and direction of the synthesis of volatile metabolites. The emission of volatile compounds was the greatest under a reduced irradiation of 322 micromoles m-2 s-1 and the smallest under 1104 micromoles m-2 s-1 at 35 degrees C. During the repair period, the contents and proportions of volatile compounds were different from their initial values, too. The degree of disruption and the following recovery of the functional state depended on the light intensity during the exposure to elevated temperatures. The investigation of the atmosphere of the growth chamber without plants has revealed the substances that were definitely technogenic in origin: tetramethylurea, dimethylsulfide, dibutylsulfide, dibutylphthalate, and a number of components of furan and silane nature.     

Significant proton events of solar cycle 22 and a comparison with events of previous solar cycles.

The sun has produced several high energy and large fluence solar proton events during solar cycle 22. This recent activity is similar to activity that occurred in the 19th solar cycle before the advent of routine space measurements. In a review of the recent events and a comparison with significant solar proton events of previous solar cycles, it appears that the 20th and 21st solar cycles were deficient in the total fluence of solar particles as detected at the earth. Therefore, when models of maximum solar proton fluxes to be encountered for deep space missions are developed, solar proton data acquired during the present solar cycle should be incorporated.     

Consistency of gas exchange of man and plants in a closed ecological system: lines of attack on the problem.

Gas exchange between man and plants in a closed ecological system based on atmosphere regeneration by plant photosynthesis is made consistent by attaining the equilibrium of human CO2 discharge and the productivity of the gas consuming bioregenerator. In this case the gas exchange might be, however, qualitatively disturbed from the equilibrium in terms of oxygen making it accumulate or decrease continuously in the air of the system. Gas exchange equilibrium in terms of O2 was attained in long-term experiments by equality of the human respiration coefficient and the plant assimilation coefficient. Varying the ratio of these parameters it is possible to control the oxygen concentration in the atmosphere to be reclaimed.     

Consideration in selecting crops for the human-rated life support system: a Linear Programming model.

A Linear Programming model has been constructed which aids in selecting appropriate crops for CELSS (Controlled Environment Life Support System) food production. A team of Controlled Environment Agriculture (CEA) faculty, staff, graduate students and invited experts representing more than a dozen disciplines, provided a wide range of expertise in developing the model and the crop production program. The model incorporates nutritional content and controlled-environment based production yields of carefully chosen crops into a framework where a crop mix can be constructed to suit the astronauts' needs. The crew's nutritional requirements can be adequately satisfied with only a few crops (assuming vitamin mineral supplements are provided) but this will not be satisfactory from a culinary standpoint. This model is flexible enough that taste and variety driven food choices can be built into the model.     

The implantation of life on Mars: feasibility and motivation.

Environmental conditions on Mars are extremely hostile, and would be destructive to any organisms which might arrive there unprotected to-day. However, it is a biocompatible planet. Its unalterable astrophysical parameters would allow the maintenance of a much thicker, warmer carbon dioxide atmosphere than that which currently exists. Though very cold (averaging about -60 degrees C), highly oxidizing and desiccated, Mars may possess substantial quantities of the materials needed to support life--in particular, water and carbon dioxide. A general scenario for implanting life on Mars would include three main phases: (1) robotic and human exploration to determine whether sufficiently large and accessible volatile inventories are available; (2) planetary engineering designed to warm the planet, release liquid water and produce a thick carbon dioxide atmosphere; and (3) if no indigenous Martian organisms emerge as liquid water becomes available, a program of biological engineering designed to construct and implant pioneering microbial communities able to proliferate in the newly clement, though still anaerobic, Martian environment. The process of establishing an ecosystem, or biosphere, on a lifeless planet is best termed 'ecopoiesis.' This new word, derived from Greek, means 'the making of an abode for life.' It is by no means clear whether ecopoiesis on Mars is scientifically possible or technologically achievable. Thus we urge that it be one of the objectives of space research during the next century to assess the feasibility of ecopoiesis on Mars.