Potential of salt-accumulating and salt-secreting halophytic plants for recycling sodium chloride in human urine in bioregenerative life support systems

This study addresses the possibility of growing different halophytic plants on mineralized human urine as a way to recycle NaCl from human wastes in a bioregenerative life support system (BLSS). Two halophytic plant species were studied: the salt-accumulating Salicornia europaea and the salt-secreting Limonium gmelinii. During the first two weeks, plants were grown on Knop’s solution, then an average daily amount of urine produced by one human, which had been preliminarily mineralized, was gradually added to the experimental solutions. Nutrient solutions simulating urine mineral composition were gradually added to control solutions. NaCl concentrations in the stock solutions added to the experimental and control solutions were 9 g/L in the first treatment and 20 g/L in the second treatment. The mineralized human urine showed some inhibitory effects on S. europaea and L. gmelinii. The biomass yield of experimental plants was lower than that of control ones. If calculated for the same time period (120 d) and area (1 m²), the amount of sodium chloride taken up by S. europaea plants would be 11.7 times larger than the amount taken up by L. gmelinii plants (486 g/m² vs. 41 g/m²). Thus, S. europaea is the better choice of halophyte for recycling sodium chloride from human wastes in BLSS.     

Crop productivities and radiation use efficiencies for bioregenerative life support

NASA’s Biomass Production Chamber (BPC) at Kennedy Space Center was decommissioned in 1998, but several crop tests were conducted that have not been reported in the open literature. These include several monoculture studies with wheat, soybean, potato, lettuce, and tomato. For all of these studies, either 10 or 20 m² of plants were grown in an atmospherically closed chamber (113 m³ vol.) using a hydroponic nutrient film technique along with elevated CO₂ (1000 or 1200 μmol mol⁻¹). Canopy light (PAR) levels ranged from 17 to 85 mol m⁻² d⁻¹ depending on the species and photoperiod. Total biomass (DM) productivities reached 39.6 g m⁻² d⁻¹ for wheat, 27.2 g m⁻² d⁻¹ for potato, 19.6 g m⁻² d⁻¹ for tomato, 15.7 g m⁻² d⁻¹ for soybean, and 7.7 g m⁻² d⁻¹ for lettuce. Edible biomass (DM) productivities reached 18.4 g m⁻² d⁻¹ for potato, 11.3 g m⁻² d⁻¹ for wheat, 9.8 g m⁻² d⁻¹ for tomato, 7.1 g m⁻² d⁻¹ for lettuce, and 6.0 g m⁻² d⁻¹ for soybean. The corresponding radiation (light) use efficiencies for total biomass were 0.64 g mol⁻¹ PAR for potato, 0.59 g DM mol⁻¹ for wheat, 0.51 g mol⁻¹ for tomato, 0.46 g mol⁻¹ for lettuce, and 0.43 g mol⁻¹ for soybean. Radiation use efficiencies for edible biomass were 0.44 g mol⁻¹ for potato, 0.42 g mol⁻¹ for lettuce, 0.25 g mol⁻¹ for tomato, 0.17 g DM mol⁻¹ for wheat, and 0.16 g mol⁻¹ for soybean. By initially growing seedlings at a dense spacing and then transplanting them to the final production area could have saved about 12 d in each production cycle, and hence improved edible biomass productivities and radiation use efficiencies by 66% for lettuce (to 11.8 g m⁻² d⁻¹ and 0.70 g mol⁻¹), 16% for tomato (to 11.4 g m⁻² d⁻¹and 0.29 g mol⁻¹), 13% for soybean (to 6.9 g m⁻² d⁻¹ and 0.19 g mol⁻¹), and 13% for potato (to 20.8 g m⁻² d⁻¹ and 0.50 g mol⁻¹). Since wheat was grown at higher densities, transplanting seedlings would not have improved yields. Tests with wheat resulted in a relatively low harvest index of 29%, which may have been caused by ethylene or other organic volatile compounds (VOCs) accumulating in the chamber. Assuming a higher harvest index of 40% could be achieved by scrubbing VOCs, productivity of wheat seed could have been improved nearly 40% to 15.8 g m⁻² d⁻¹ and edible biomass radiation use efficiency to 0.30 g mol⁻¹.     

Aquatic food production modules in bioregenerative life support systems based on higher plants.

Most bioregenerative life support systems (BLSS) are based on gravitropic higher plants which exhibit growth and seed generation disturbances in microgravity. Even when used for a lunar or martian base the reduced gravity may induce a decreased productivity in comparison to Earth. Therefore, the implementation of aquatic biomass production modules in higher plant and/or hybrid BLSS may compensate for this and offer, in addition, the possibility to produce animal protein for human nutrition. It was shown on the SLS-89 and SLS-90 space shuttle missions with the C.E.B.A.S.-MINI MODULE that the edible non gravitropic rootless higher aquatic plant Ceratophyllum demeresum exhibits an undisturbed high biomass production rate in space and that the teleost fish species, Xiphophorus helleri, adapts rapidly to space conditions without loss of its normal reproductive functions. Based on these findings a series of ground-based aquatic food production systems were developed which are disposed for utilization in space. These are plant production bioreactors for the species mentioned above and another suitable candidate, the lemnacean (duckweed) species, Wolffia arrhiza. Moreover, combined intensive aquaculture systems with a closed food loop between herbivorous fishes and aquatic and land plants are being developed which may be suitable for integration into a BLSS of higher complexity. Grant numbers: WS50WB9319-3, IVA1216-00588.     

Plants survive rapid decompression: Implications for bioregenerative life support

Radish (Raphanus sativus), lettuce (Latuca sativa), and wheat (Triticum aestivum) plants were grown at either 98 kPa (ambient) or 33 kPa atmospheric pressure with constant 21 kPa oxygen and 0.12 kPa carbon dioxide in atmospherically closed pressure chambers. All plants were grown rockwool using recirculating hydroponics with a complete nutrient solution. At 20 days after planting, chamber pressures were pumped down as rapidly as possible, reaching 5 kPa after about 5 min and ∼1.5 kPa after about 10 min. The plants were held at 1.5 kPa for 30 min and then pressures were restored to their original settings. Temperature (22 °C) and humidity (65% RH) controls were engaged throughout the depressurization, although temperatures dropped to near 16 °C for a brief period. CO₂ and O₂ were not detectable at the low pressure, suggesting that most of the 1.5 kPa atmosphere consisted of water vapor. Following re-pressurization, plants were grown for another 7 days at the original pressures and then harvested. The lettuce, radish, and wheat plants showed no visible effects from the rapid decompression, and there were no differences in fresh or dry mass when compared to control plants maintained continuously at 33 or 98 kPa. But radish storage root fresh mass and lettuce head fresh and dry masses were less at 33 kPa compared to 98 kPa for both the controls and decompression treatment. The results suggest that plants are extremely resilient to rapid decompression, provided they do not freeze (from evaporative cooling) or desiccate. The water of the hydroponic system was below the boiling pressure during these tests and this may have protected the plants by preventing pressures from dropping below 1.5 kPa and maintaining humidity near 1.5 kPa. Further testing is needed to determine how long plants can withstand such low pressure, but the results suggest there are at least 30 min to respond to catastrophic pressure losses in a plant production chamber that might be used for life support in space.     

Gas exchange rates of potato stands for bioregenerative life support

Plants can provide a means for removing carbon dioxide (CO₂) while generating oxygen (O₂) and clean water for life support systems in space. To study this, 20 m² stands of potato (Solanum tuberosum L.) plants were grown in a large (113 m³ vol.), atmospherically closed chamber. Photosynthetic uptake of CO₂ by the stands was detected about 10 DAP (days after planting), after which photosynthetic rates rose rapidly as stand ground cover and total light interception increased. Photosynthetic rates peaked ca. 50 DAP near 45 μmol CO₂ m⁻² s⁻¹ under 865 μmol m⁻² s⁻¹ PPF (average photosynthetic photon flux), and near 35 μmol CO₂ m⁻² s⁻¹ under 655 μmol m⁻² s⁻¹ PPF. Short term changes in PPF caused a linear response in stand photosynthetic rates up to 1100 μmol m⁻² s⁻¹ PPF, with a light compensation point of 185 μmol m⁻² s⁻¹ PPF. Comparisons of stand photosynthetic rates at different CO₂ concentrations showed a classic C₃ response, with saturation occurring near 1200 μmol mol⁻¹ CO₂ and compensation near 100 μmol mol⁻¹ CO₂. In one study, the photoperiod was changed from 12 h light/12 h dark to continuous light at 58 DAP. This caused a decrease in net photosynthetic rates within 48 h and eventual damage (scorching) of upper canopy leaves, suggesting the abrupt change stressed the plants and/or caused feedback effects on photosynthesis. Dark period (night) respiration rates increased during early growth as standing biomass increased and peaked near 9 μmol CO₂ m⁻² s⁻¹ ca. 50 DAP, after which rates declined gradually with age. Stand transpiration showed a rapid rise with canopy ground cover and peaked ca. 50 DAP near 8.9 L m⁻² d⁻¹ under 860 μmol m⁻² s⁻¹ PPF and near 6.3 L m⁻² d⁻¹ under 650 μmol m⁻² s⁻¹ PPF. Based on the best photosynthetic rates from these studies, approximately 25 m² of potato plants under continuous cultivation would be required to support the CO₂ removal and O₂ requirements for one person.     

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.     

Material recycling in a regenerative life support system for space use: its issues and waste processing.

First, sources of wastes produced in future human activities in space will be discussed. The quantity and quality of each source will be evaluated. Secondly, water quality requirements for human safety will be shown. Then, techniques to be used for processing and/or recycling the wastes to attain the requirements will be listed. Specific characteristics and limitations of each will be explained. Last, examples of system configurations of the techniques will be shown. The material balances will also be calculated. The issues to be solved will be made clear.     

Material balance and diet in bioregenerative life support systems: connection with coefficient of closure.

Bioregenerative life support systems (BLSS) with different coefficients of closure are considered. The 66.2% coefficient of closure achieved in "BIOS-3" facility experiments has been taken as a base value. The increase in coefficient of closure up to 72.6-93.0% is planned due to use of soil-like substrate (SLS) and concentrating of urine. Food values were estimated both in a base variant ("BIOS-3"), and with increases in the coefficient of closure. It is shown that food requirements will be more fully satisfied by internal crop production with an increase in the coefficient of closure of the BLSS. Changes of massflow rates on an 'input-output' and inside BLSS are considered. Equations of synthesis and degradation of organic substances in BLSS were examined using a stoichiometric model. The paper shows that at incomplete closure of BLSS containing SLS there is a problem of nitrogen balancing. To compensate for the removal of nitrogen from the system in urine and feces, it is necessary to introduce food and a nitrogen-containing additive.     

Possible applications of aquatic bioregenerative life support modules for food production in a Martian base.

Water is the essential precondition of life in general and also for the establishment of a Martian base suitable for long duration stays of humans. It is not yet proven if there is indeed a "frozen ocean" under the surface of Mars but if this could be verified it would open innovative aspects for the construction of bioregenerative life support systems (BLSS). In a general concept higher plants will play the predominant role in a Martian BLSS. It is not clear, however, how these will grow and bring seed in reduced gravity and there may be differences in the productivity in comparison to Earth conditions. Therefore, organisms which are already adapted to low gravity conditions, namely non-gravitropic aquatic plants and also aquatic animals may be used to enhance the functionality of the Martian BLSS as a whole. It has been shown already with the so-called C.E.B.A.S. MINIMODULE in the STS-89 and STS-90 spaceshuttle missions that the water plant Ceratophyllum demersum has an undisturbed and high biomass production under space conditions. Moreover, the teleost fish species Xiphophorus helleri adapted easily to the micro-g environment and maintained its normal reproductive functions. Based on this findings a possible scenario is presented in which aquatic plant production modules and combined animal-plant production systems may be used for human food production and water and air regeneration in a Martian base.     

Selection and hydroponic growth of potato cultivars for bioregenerative life support systems

As part of the ESA-funded MELiSSA program, Ghent University and the Université catholique de Louvain investigated the suitability, growth and development of four potato cultivars in hydroponic culture under controlled conditions with the aim to incorporate such cultivation system in an Environmental Control and Life Support System (ECLSS). Potato plants can fulfill three major functions in an ECLSS in space missions: (a) fixation of CO₂ and production of O₂, (b) production of tubers for human nutrition and (c) production of clean water after condensation of the water vapor released from the plants by transpiration. Four cultivars (Annabelle, Bintje, Desiree and Innovator) were selected and grown hydroponically in nutrient film technique (NFT) gullies in a growth chamber under controlled conditions. The plant growth parameters, tuber harvest parameters and results of tuber nutritional analysis of the four cultivars were compared. The four potato cultivars grew well and all produced tubers. The growth period lasted 127 days for all cultivars except for Desiree which needed 145 days. Annabelle (1.45 kg/m²) and Bintje (1.355 kg/m²) were the best performing of the four cultivars. They also produced two times more tubers than Desiree and Innovator. Innovator produced the biggest tubers (20.95 g/tuber) and Desiree the smallest (7.67 g/tuber). The size of Annabelle and Bintje potatoes were intermediate. Bintje plants produced the highest total biomass in term of DW. The highest non-edible biomass was produced by Desiree, which showed both the highest shoot and root DW. The manual length and width measurements were also used to predict the total tuber mass. The energy values of the tubers remained in the range of the 2010 USDA and Souci-Fachmann-Kraut food composition databases. The amount of Ca determined was slightly reduced compared to the USDA value, but close to the Souci-Fachmann-Kraut value. The concentration of Cu, Zn and P were high compared to both databases.     

Conceptual design of a bioregenerative life support system containing crops and silkworms

This article summarizes a conceptual design of a bioregenerative life support system for permanent lunar base or planetary exploration. The system consists of seven compartments – higher plants cultivation, animal rearing, human habitation, water recovery, waste treatment, atmosphere management, and storages. Fifteen kinds of crops, such as wheat, rice, soybean, lettuce, and mulberry, were selected as main life support contributors to provide the crew with air, water, and vegetable food. Silkworms fed by crop leaves were designated to produce partial animal nutrition for the crew. Various physical-chemical and biological methods were combined to reclaim wastewater and solid waste. Condensate collected from atmosphere was recycled into potable water through granular activated carbon adsorption, iodine sterilization, and trace element supplementation. All grey water was also purified though multifiltration and ultraviolet sterilization. Plant residue, human excrement, silkworm feces, etc. were decomposed into inorganic substances which were finally absorbed by higher plants. Some meat, ingredients, as well as nitrogen fertilizer were prestored and resupplied periodically. Meanwhile, the same amount and chemical composition of organic waste was dumped to maintain the steady state of the system. A nutritional balanced diet was developed by means of the linear programming method. It could provide 2721 kcal of energy, 375.5 g of carbohydrate, 99.47 g of protein, and 91.19 g of fat per capita per day. Silkworm powder covered 12.54% of total animal protein intakes. The balance of material flows between compartments was described by the system of stoichiometric equations. Basic life support requirements for crews including oxygen, food, potable and hygiene water summed up to 29.68 kg per capita per day. The coefficient of system material closure reached 99.40%.     

Selection and hydroponic growth of bread wheat cultivars for bioregenerative life support systems

As part of the ESA-funded MELiSSA program, the suitability, the growth and the development of four bread wheat cultivars were investigated in hydroponic culture with the aim to incorporate such a cultivation system in an Environmental Control and Life Support System (ECLSS). Wheat plants can fulfill three major functions in space: (a) fixation of CO₂ and production of O₂, (b) production of grains for human nutrition and (c) production of cleaned water after condensation of the water vapor released from the plants by transpiration. Four spring wheat cultivars (Aletsch, Fiorina, Greina and CH Rubli) were grown hydroponically and compared with respect to growth and grain maturation properties. The height of the plants, the culture duration from germination to harvest, the quantity of water used, the number of fertile and non-fertile tillers as well as the quantity and quality of the grains harvested were considered. Mature grains could be harvested after around 160 days depending on the varieties. It became evident that the nutrient supply is crucial in this context and strongly affects leaf senescence and grain maturation. After a first experiment, the culture conditions were improved for the second experiment (stepwise decrease of EC after flowering, pH adjusted twice a week, less plants per m²) leading to a more favorable harvest (higher grain yield and harvest index). Considerably less green tillers without mature grains were present at harvest time in experiment 2 than in experiment 1. The harvest index for dry matter (including roots) ranged from 0.13 to 0.35 in experiment 1 and from 0.23 to 0.41 in experiment 2 with modified culture conditions. The thousand-grain weight for the four varieties ranged from 30.4 to 36.7 g in experiment 1 and from 33.2 to 39.1 g in experiment 2, while market samples were in the range of 39.4–46.9 g. Calcium levels in grains of the hydroponically grown wheat were similar to those from field-grown wheat, while potassium, magnesium, phosphorus, iron, zinc, copper, manganese and nickel levels tended to be higher in the grains of experimental plants. It remains a challenge for future experiments to further adapt the nutrient supply in order to improve senescence of vegetative plant parts, harvest index and the composition of bread wheat grains.     

Comparison of three soil-like substrate production techniques for a bioregenerative life support system

It is very important to recycle the inedible biomass of higher plants to improve the closure of bioregenerative life support system (BLSS). Processing candidate higher plant residues into the soil-like substrate (SLS) as the plant growth medium is a promising way to achieve. In this study, three different processing techniques of SLSs, using residues of wheat and rice as feedstock, were compared. As for the first traditional technique, SLS1 was obtained by successive conversion of wheat straw by oyster mushrooms and worms. In the other two methods, SLSs were produced with aerobic fermentation (SLS2) or anaerobic fermentation (SLS3) followed by worm conversion. The changes in SLS cellulose, lignin, available elements and pH were measured during the production processes. The maturity was evaluated by the value of C/N. The fertilities were compared in terms of available elements contents and lettuce productivities. The results indicated that the second technique was optimal, whose process cycle was 30 days less than that of SLS1. The total cellulose and lignin degradation of SLS2, achieved 98.6% and 93.1% during the 93-days-processing, and the lettuce productivity reached 12.0 g m⁻² day⁻¹.     

Testing anti-fungal activity of a soil-like substrate for growing plants in bioregenerative life support systems

The object of this research is to study a soil-like substrate (SLS) to grow plants in a Bioregenerative Life Support System (BLSS). Wheat and rice straw were used as raw materials to prepare SLS. Anti-fungal activity of SLS using test cultures of Bipolaris sorokiniana, a plant-pathogenic fungus which causes wheat root rot was studied. Experiments were conducted with SLS samples, using natural soil and sand as controls. Infecting the substrates, was performed at two levels: the first level was done with wheat seeds carrying B. sorokiniana and the second level with seeds and additional conidia of B. sorokiniana from an outside source. We measured wheat disease incidence and severity in two crop plantings. Lowest disease incidence values were obtained from the second planting, SLS: 26% and 41% at the first and the second infection levels, respectively. For soil the values were 60% and 82%, respectively, and for sand they were 67% and 74%, respectively. Wheat root rot in the second crop planting on SLS, at both infection levels was considerably less severe (9% and 13%, respectively) than on natural soil (20% and 33%) and sand (22% and 32%). SLS significantly suppressed the germination of B. sorokiniana conidia. Conidia germination was 5% in aqueous SLS suspension, and 18% in clean water. No significant differences were found regarding the impact on conidia germination between the SLS samples obtained from wheat and rice straw. The anti-fungal activity in SLS increased because of the presence of worms. SLS also contained bacteria stimulating and inhibiting B. sorokiniana growth.     

Manipulating light and temperature to minimize environmental stress in the plant component of bioregenerative life support systems.

Our experiments examined enhancing tolerance of the photosynthesizing component to possible deviations in thermal or illumination conditions inside a bioregenerative life support system (BLSS). In the event of one parameter getting beyond its optimum, the values of other parameters may ensure minimal damage to the plant component during the period of environmental stress. With wheat plants (one of key elements of the plant component) as an example the work considers whether it is possible to enhance thermal tolerance by varying light intensity. Increase of air temperature to 35 degrees C or 45 degrees C with light intensity of 60 W/m2 PAR has been shown to substantially inhibit the photosynthesis processes; at 150 W/m2 PAR photosynthesis decreases from 50% to 100%, respectively; when light intensity is increased to 240 W/m2 PAR photosynthesis increased more than 70% at 35 degrees C and decreased at 45 degrees C by only 20%. Thus, light intensity can be increased to avoid or decrease the inhibiting effect of high temperatures. On the other hand, tolerance of wheat plants to prolonged absence of light can be substantially enhanced by decreasing during this period air temperature to temperatures close to 0 degrees C.     

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.     

Carbon balance in bioregenerative life support systems: some effects of system closure, waste management, and crop harvest index.

In Advanced Life Support (ALS) systems with bioregenerative components, plant photosynthesis would be used to produce O2 and food, while removing CO2. Much of the plant biomass would be inedible and hence must be considered in waste management. This waste could be oxidized (e.g., incinerated or aerobically digested) to resupply CO2 to the plants, but this would not be needed unless the system were highly closed with regard to food. For example, in a partially closed system where some of the food is grown and some is imported, CO2 from oxidized waste when combined with crew and microbial respiration could exceed the CO2 removal capability of the plants. Moreover, it would consume some O2 produced from photosynthesis that could have been used by the crew. For partially closed systems it would be more appropriate to store or find other uses for the inedible biomass and excess carbon, such as generating soils or growing woody plants (e.g., dwarf fruit trees). Regardless of system closure, high harvest crops (i.e., crops with a high edible to total biomass ratio) would increase food production per unit area and O2 yields for systems where waste biomass is oxidized to recycle CO2. Such interlinking effects between the plants and waste treatment strategies point out the importance of oxidizing only that amount of waste needed to optimize system performance.     

Waste bioregeneration in life support CES: development of soil organic substrate.

An experimental model of matter turnover in the biotic cycle: plants (plant biomass) --> mushrooms (residual substrate + mushroom fruit bodies) --> worms (biohumus) --> microorganisms (soillike substrate) --> plants is presented. The initial mass of soillike substrate was produced from wheat plants grown in a hydroponic system. Three cycles of matter turnover in the biotic cycle were carried out. Grain productivity on soillike substrate was 21.87 g/m2 day. The results obtained were used for designing a CES containing man, plants, soillike substrate, bioregeneration module and aquaculture. It was shown, that the circulating dry mass of the CES is 756 kg. The main part (88%) of the circulating mass accumulates in the soillike substrate and bioregeneration module.     

Effect of salt stress on growth and physiology in amaranth and lettuce: Implications for bioregenerative life support system

Growing plants can be used to clean waste water in bioregenerative life support system (BLSS). However, NaCl contained in the human urine always restricts plant growth and further reduces the degree of mass cycle closure of the system (i.e. salt stress). This work determined the effect of NaCl stress on physiological characteristics of plants for the life support system. Amaranth (Amaranthus tricolor L. var. Huahong) and leaf lettuce (Lactuca sativa L. var. Luoma) were cultivated at nutrient solutions with different NaCl contents (0, 1000, 5000 and 10,000 ppm, respectively) for 10 to 18 days after planted in the Controlled Ecological Life Support System Experimental Facility in China. Results showed that the two plants have different responses to the salt stress. The amaranth showed higher salt-tolerance with NaCl stress. If NaCl content in the solution is below 5000 ppm, the salt stress effect is insignificant on above-ground biomass output, leaf photosynthesis rate, Fv/Fm, photosynthesis pigment contents, activities of antioxidant enzymes, and inducing lipid peroxidation. On the other hand, the lettuce is sensitive to NaCl which significantly decreases those indices of growth and physiology. Notably, the lettuce remains high productivity of edible biomass in low NaCl stress, although its salt-tolerant limitation is lower than amaranth. Therefore, we recommended that amaranth could be cultivated under a higher NaCl stress condition (<5000 ppm) for NaCl recycle while lettuce should be under a lower NaCl stress (<1000 ppm) for water cleaning in future BLSS.     

Chlorella vulgaris culture as a regulator of CO2 in a bioregenerative life support system

It is the primary task for a bioregenerative life support system (BLSS) to maintain the stable concentrations of CO₂ and O₂. However, these concentrations could fluctuate based on various factors, such as the imbalance between respiration/assimilation quotients of the heterotrophic and autotrophic components. They can even be out of balance through catastrophic failure of higher plants in the emergency conditions. In this study, the feasibility of using unicellular Chlorella vulgaris of typically rapid growth as both “compensatory system” and “regulator” to control the balance of CO₂ and O₂ was analyzed in a closed ecosystem. For this purpose, a small closed ecosystem called integrative experimental system (IES) was established in our laboratory where we have been conducting multi-biological life support system experiments (MLSSE). The IES consists of a closed integrative cultivating system (CICS) and a plate photo-bioreactor. Four volunteers participated in the study for gas exchange by periodical breathing through a tube connected with the CICS. The plate photo-bioreactor was used to cultivate C. vulgaris. Results showed that the culture of C. vulgaris could be used in a situation of catastrophic failure of higher plant under the emergencies. And the productivity could recover itself to the original state in 3 to 5 days to protect the system till the higher plant was renewed. Besides, C. vulgaris could grow well and the productivity could be affected by the light intensity which could help to keep the balance of CO₂ and O₂ in the IES efficiently. Thus, C. vulgaris could be included in the design of a BLSS as a “compensatory system” in the emergency contingency and a “regulator” during the normal maintenance.