Food production and gas exchange system using blue-green alga (Spirulina) for CELSS.

In order to reduce the cultivation area required for the growth of higher plants in space adoption of algae, which have a higher photosynthetic ability, seems very suitable for obtaining oxygen and food as a useful source of high quality protein. The preliminary cultivation experiment for determining optimum cultivation conditions and for obtaining the critical design parameters of the cultivator itself has been conducted. Spirulina was cultivated in the 6-liter medium containing sodium hydrogen carbonate solution and a cultivation temperature controlled using a thermostat. Generated oxygen gas was separated using a polypropyrene porous hollow fiber membrane module. Through this experiment, oxygen gas (at a concentration of more than 46%) at a rate of 100-150 ml per minute could be obtained.     

Closed and continuous algae cultivation system for food production and gas exchange in CELSS.

In CELSS (Controlled Ecological Life Support System), utilization of photosynthetic algae is an effective means for obtaining food and oxygen at the same time. We have chosen Spirulina, a blue-green alga, and have studied possibilities of algae utilization. We have developed an advanced algae cultivation system, which is able to produce algae continuously in a closed condition. Major features of the new system are as follows. (1) In order to maintain homogeneous culture conditions, the cultivator was designed so as to cause a swirl on medium circulation. (2) Oxygen gas separation and carbon dioxide supply are conducted by a newly designed membrane module. (3) Algae mass and medium are separated by a specially designed harvester. (4) Cultivation conditions, such as pH, temperature, algae growth rate, light intensity and quantity of generated oxygen gas are controlled by a computer system and the data are automatically recorded. This equipment is a primary model for ground experiments in order to obtain some design data for space use. A feasibility of algae cultivation in a closed condition is discussed on the basis of data obtained by use of this new system.     

Development of a ground-based space micro-algae photo-bioreactor

The purpose of the research is to develop a photo-bioreactor which may produce algae protein and oxygen for future astronauts in comparatively long-term exploration, and remove carbon dioxide in a controlled ecological life support system. Based on technical parameters and performance requirements, the project planning, design drafting, and manufacture were conducted. Finally, a demonstration test for producing algae was done. Its productivity for micro-algae and performance of the photo-bioreactor were evaluated. The facility has nine subsystems, including the reactor, the illuminating unit, the carbon dioxide (CO₂) production unit and oxygen (O₂) generation unit, etc. The demonstration results showed that the facility worked well, and the parameters, such as energy consumption, volume, and productivity for algae, met with the design requirement. The density of algae in the photo-bioreactor increased from 0.174 g (dry weight) L⁻¹ to 4.064 g (dry weight) L⁻¹ after 7 days growth. The principle of providing CO₂ in the photo-bioreactor for algae and removing O₂ from the culture medium was suitable for the demand of space conditions. The facility has reasonable technical indices, and smooth and dependable performances.     

Sunlight supply and gas exchange systems in the microalgal bioreactor.

The bioreactor with sunlight supply system and gas exchange systems presented here has proved feasible in ground tests and shows much promise for space use as a CELSS device. Our chief conclusions concerning the specification of total system needed for a life support system for a man in a space station are the following. (1) Sunlight supply system: compactness and low electrical consumption. (2) Bioreactor system: high density and growth rate of chlorella. (3) Gas exchange system: enough for O2 production and CO2 assimilation.     

Competition between links in "producer-consumer" trophic chains in an aquatic closed system with spatially separated components.

The work analyzes functioning of a "producer-consumer" closed aquatic system with spatially separated links, where each component consisted of two species. Producers in the system were the microalgae of Chlorella vulgaris and Scenedesmus sp., consumers--Paramecium caudatum infusoria and Brachionus sp. rotifers. In the experiment the competing predators were consuming on a mixed culture of algae, and the competition of algae was studied under nitrogen limitation. Under these conditions competitiveness of Scenedesmus was higher than that of Chlorella vulgaris. Metabolism products of Scenedesmus algae have been found to have negative effect on reproduction of Paramecium caudatum protozoa. Predator population dynamics in the "consumer" link demonstrated that the rotifers that consume two algal species are more competitive compared to protozoa feeding on chlorella only. Grant numbers: N99-04-96017, N25.     

Operation of an experimental algal gas exchanger for use in a CELSS.

Concepts of a CELSS anticipate the use of photosynthetic organisms (higher plants and algae) for air revitalization. The rates of production and uptake of carbon dioxide and oxygen between the crew and the photosynthetic organisms are mismatched. An algal [correction of aglal] system used for gas exchange only will have the difficulty of an accumulation or depletion of these gases beyond physiologically tolerable limits (in a materially closed system the mismatch between assimilatory quotient (AQ) and respiratory quotient (RQ) will be balanced by the operation of the waste processor). We report the results of a study designed to test the feasibility of using environmental manipulations to maintain physiologically appropriate atmospheres for algae (Chlorella pyrenoidosa) and mice (Mus musculus strain DW/J) in a gas-closed system. Specifically, we consider the atmosphere behavior of this system with Chlorella grown on nitrate or urea and at different light intensities and optical densities. Manipulation of both the photosynthetic rate and AQ of the alga has been found to reduce the mismatch of gas requirements and allow operation of the system in a gas-stable manner. Operation of such a system in a CELSS may be useful for reduction of buffer sizes, as a backup system for higher plant air revitalization and to supply extra oxygen to the waste processor or during crew changes. In addition, mass balance for components of the system (mouse, algae and a waste processor) are presented.     

地外人工光合成装置研制与试验

原位资源利用技术是地外生命保障体系构建、实现人类地外生存的有效途径,是载人深空探索的核心技术。基于微通道技术的人工光合成反应器,采用流动反应器设计,用于低微重力等特殊环境条件下模拟人工光合作用,实现CO２向O２和含碳燃料的转化。微通道芯片通过气液剪切作用力使气体反应产物快速脱离电极表面并随反应介质排出反应器,理论上可以克服微重力条件对反应过程的影响,尚需进行微重力试验进行验证。同时,微通道结构可以通过精确控制反应气液的压力、流速、流量比等反应条件,获得优化的反应条件。通过地面试验,验证了该反应器将CO２还原为O２和含碳化合物的功能可行性。以Au和Ir/C作为阴极和阳极材料,3V电压条件下,O２产率可达11.74mL/h。此外,基于人工光合成反应器搭建了集反应模块、控制模块、流路驱动模块以及检测模块等于一体的地外人工光合成装置,形成原位反应、介质供给、精确控制、在线收集和检测等功能一体化的系统,并实现CO２有效转换和O２供给。为后续技术成熟度更高的反应装置研制、高产物选择性的含碳化合物转化以及人工光合成反应装置在轨试验奠定了理论和实践基础。     

Atmosphere behavior in gas-closed mouse-algal systems: An experimental and modelling study

Concepts of biologically-based regenerative life support systems anticipate the use of photosynthetic organisms for air revitalization. However, mismatches in the rates of production and uptake of oxygen or carbon dioxide between the crew and the plants will lead to an accumulation or depletion of these gases beyond tolerable limits. One method for correcting these atmospheric changes is to use physicochemical devices. This would conflict with the constraint of minimal size and weight imposed upon the successful development of a competitive bioregenerative system. An alternate control strategy is based upon reducing the gas exchange mismatch by manipulation of those environmental parameters known to affect plant or algae gas exchange ratios. We have initiated a research program using a dual approach of mathematical modelling and laboratory experimentation aimed at examining the gas exchange characteristics of artificial animal/plant systems closed to the ambient atmosphere. Our goal is to develop control techniques and management strategies for maintaining the atmospheric levels of carbon dioxide and oxygen at physiological levels. A mathematical model simulating the atmospheric behavior in these systems has been developed and an experimental gas-closed system has been constructed. These will be described and preliminary results will be presented.     

Spaceflight hardware allowing unilateral irradiation and chemical fixation in petri dishes.

To accommodate a spaceflight experiment with moss (SPM), experiment-unique equipment (EUE) was developed by engineers at Kennedy Space Center. The hardware allows sterile culture for an extended period of time in commercial petri dishes, lateral illumination of each culture with light of a specific wavelength (660 nm; other wavelengths are possible) and a range of intensities (0.05-5 micromoles photons m-2 s-1), incubation in complete darkness, and chemical fixation to terminate the experiment under conditions of microgravity. The use of a fixative required triple containment to protect the astronaut crew. An external panel on the experiment container allowed the timing of illumination and fixation to be controlled by the crew. Light quality is provided by light emitting diodes (LEDs) that are located in the lid of the outer container, the BRIC (Biological Research In Canisters)-LED. Each canister accommodates 6 Petri Dish Fixation Units (PDFUs), and each PDFU holds one 6 cm petri dish. All components are autoclavable. LED illumination is piped through a transparent glass rod. Each PDFU contains fixative in a reservoir that is released by the depression of an actuator. This hardware performed well during its first flight, the 16-day STS-87 mission in Nov./Dec., 1997 as part of the Collaborative USA and Ukrainian Experiment (CUE). It supported vigorous and sterile moss growth, cells were maintained in position and were well-fixed, and there was a vigorous and consistent response to light. Although here used for moss, in future flight experiments this unique new hardware can be used for many types of organisms normally grown in petri dishes, with or without a requirement for illumination.     

Nostoc sphaeroides Kützing,an excellent candidate producer for CELSS

Some phytoplankton can be regarded as possible candidates in the establishment of Controlled Ecological Life Support System (CELSS) for some intrinsic characteristics, the first characteristic is that they should grow rapidly, secondly, they should be able to endure some stress factors and develop some corresponding adaptive strategies; also it is very important that they could provide food rich in nutritious protein and vitamins for the crew; the last but not the least is they can also fulfill the other main functions of CELSS, including supplying oxygen, removing carbon dioxide and recycling the metabolic waste. According to these characteristics, Nostoc sphaeroides, a potential healthy food in China, was selected as the potential producer in CELSS. It was found that the oxygen average evolution rate of this algae is about 150 μmol O₂ mg⁻¹ h⁻¹, and the size of them are ranged from 2 to 20 mm. Also it can be cultured with high population density, which indicated that the potential productivity of Nostoc sphaeroides is higher than other algae in limited volume. We measured the nutrient contents of the cyanobacterium and concluded it was a good food for the crew. Based on above advantages, Nostoc sphaeroides was assumed to a suitable phytoplankton for the establishment of Controlled Ecological Life Support System. We plan to develop suitable bioreactor with the cyanobacterium for supplying oxygen and food in future space missions.     

LED光谱对模拟空间培养箱中植物生长发育的影响

通过研究在空间植物培养箱中利用LED作为光源对植物生长发育的作用, 并以荧光灯作为对照, 评估LED光源在空间植物培养中的优缺点, 可为中国即将在空间实验室天宫二 号和空间站中开展的高等植物生长实验提供参考. 利用不同比例的红光与白光LED组合光源, 研究光谱组成(红蓝光比例)、光照强度、光周期和气体流通等条件对于模拟空间 植物培养箱中拟南芥和水稻生长发育的影响. 结果表明, 与荧光灯相比, 红蓝光比例高的LED会导致拟南芥提早开花和水稻叶片的早衰. 红蓝光峰值比在3.9左右时, 拟南芥 和水稻生长最为有利; 红蓝光峰值比超过16则明显抑制拟南芥和水稻的生长, 导致叶片早衰. 另外, 在密闭培养箱中, 光强小于150μmol·m-2·s-1时, 增加光照强度可以部分抵消气体流通不足导致拟南芥植物生长的抑制, 而光照强度大 于150μmol·m-2·s-1时, 光强越大拟南芥的生长发育受到抑制越严重. 水稻对密闭培养环境中高光强的耐受性明显好于拟南芥. 因此, 在设计空间植物培养箱的LED光照系统时, 红蓝光的比例选择是关键, 此外还需综合考虑空间微重力条件下气体对流变化影响植物对光的反应.      

Mass exchange in an experimental new-generation life support system model based on biological regeneration of environment.

An experimental model of a biological life support system was used to evaluate qualitative and quantitative parameters of the internal mass exchange. The photosynthesizing unit included the higher plant component (wheat and radish), and the heterotrophic unit consisted of a soil-like substrate, California worms, mushrooms and microbial microflora. The gas mass exchange involved evolution of oxygen by the photosynthesizing component and its uptake by the heterotroph component along with the formation and maintaining of the SLS structure, growth of mushrooms and California worms, human respiration, and some other processes. Human presence in the system in the form of "virtual human" that at regular intervals took part in the respirative gas exchange during the experiment. Experimental data demonstrated good oxygen/carbon dioxide balance, and the closure of the cycles of these gases was almost complete. The water cycle was nearly 100% closed. The main components in the water mass exchange were transpiration water and the watering solution with mineral elements. Human consumption of the edible plant biomass (grains and roots) was simulated by processing these products by a unique physicochemical method of oxidizing them to inorganic mineral compounds, which were then returned into the system and fully assimilated by the plants. The oxidation was achieved by "wet combustion" of organic biomass, using hydrogen peroxide following a special procedure, which does not require high temperature and pressure. Hydrogen peroxide is produced from the water inside the system. The closure of the cycle was estimated for individual elements and compounds. Stoichiometric proportions are given for the main components included in the experimental model of the system. Approaches to the mathematical modeling of the cycling processes are discussed, using the data of the experimental model. Nitrogen, as a representative of biogenic elements, shows an almost 100% closure of the cycle inside the system. The proposed experimental model of a biological system is discussed as a candidate for potential application in the investigations aimed at creating ecosystems with largely closed cycles of the internal mass exchange. The formation and maintenance of sustainable cycling of vitally important chemical elements and compounds in biological life support systems (BLSS) is an extremely pressing problem. To attain the stable functioning of biological life support systems (BLSS) and to maintain a high degree of closure of material cycles in than, it is essential to understand the character of mass exchange processes and stoichiometnc proportions of the initial and synthesized components of the system.     

Instrumentation for plant health and growth in space.

The present-day plant growth facilities ("greenhouses") for space should be equipped with monitors and controllers of ambient parameters within the chamber because spacecraft environmental variations can be unfavorable to plants. Moreover, little is known about the effects of spaceflight on the greenhouse and rooting media. Lack of information about spaceflight effects on plants necessitates supplying space greenhouses with automatic, non-invasive monitors of, e.g., gas exchange rate, water and nutrient ion uptake, plant mass, temperature and water content of leaves. However, introduction of an environmental or plant sensor into the monitoring system may be reasonable only if it is justified by quantitative evaluation of the influence of a measured parameter on productivity, efficacy of illumination, or some other index of greenhouse efficiency. The multivariate adaptive optimization in terrestrial phytotrons appears to be one of the best methods to assess environmental impacts on crops. Two modifications of greenhouses with the three-dimensional adaptive optimization of crop photosynthetic characteristics include: (1) irradiation, air temperature and carbon dioxide using a modified simplex algorithm; and (2) using irradiation, air temperature, and humidity with sensitivity algorithms with varying frequency of test exposures that have been experimentally developed. As a result, during some stages of plant ontogensis, the photosynthetic productivity of wheat, tomatoes, and Chinese cabbage in these systems was found to increase by a factor of 2-3.     

Experiment "Seeds" on Biokosmos 9. Dosimetric part.

The aim of the experiment "Seeds" on the Sowjetic satellite Biokosmos 9 was the observation of mutagenic effects caused at special loci of seeds of Arabidopsis thaliana and assigned to particles of the Cosmic radiation. Two types of exposure units were flown: A low-shielding unit Type I, mounted at the surface of the satellite (1.4 g/cm2 shielding) and, for comparison, an identical item inside (16 g/cm2 shielding), using nuclear emulsion as track detectors. A Type II unit, flown inside (18g/cm2 shielding) was mounted with AgCl track detectors. The layout will be briefly described. A first set of dosimetric data from the physical evaluation of the experiment will be presented. The subdivision into charge- and LET-groups shows a rather high contribution of the intermediate LET-group (350-1000 MeV/cm) due to medium heavy particles (Z = 6-10) and to enders of light (p, alpha) particles.     

Growth of plants at reduced pressures: experiments in wheat-technological advantages and constraints.

Procedures and results are presented concerning the growth of wheat plants with variable partial pressures of O2 and N2. Data demonstrate that some growth occurs in pressures as low as 0.1 atmosphere. The growth was similar or higher at 200 mb (0.2 atmosphere) than in normal atmosphere but the development was different. Advantages of the low pressure cultivation, especially in the absence of nitrogen, are discussed, including better ratio volume/mass of plant cultivation module; lower losses of gases by leakage; easier management of photosynthetic oxygen produced by plants.     

A straw-soil co-composting and evaluation for plant substrate in BLSS

Material closure is important for the establishment of Bioregenerative Life Support System, and many studies have focused on transforming candidate plant residues into plant culture medium. For the limitations of using wheat straw compost as substrate for plant cultivation, a straw-soil co-composting technique was studied. The changes of pH, C/N value, germination index, cellulose, lignin and so on were monitored during the co-composting process. The maturity was evaluated by the C/N value and the germination index. The result showed that after 45 days’ fermentation, the straw-soil final co-compost with inoculation (T1) became mature, while the co-compost without inoculation (T0) was not mature. In the plant culture test, the T1 substrate could satisfy the needs for lettuce’s growth, and the edible biomass yield of lettuce averaged 74.42 g pot⁻¹ at harvest. But the lettuces in T0 substrate showed stress symptoms and have not completed the growth cycle. Moreover, the results of nitrogen (N) transformation experiment showed that about 10.0% and 3.1% N were lost during the T1 co-composting and plant cultivation, respectively, 23.5% N was absorbed by lettuce, and 63.4% N remained in the T1 substrate after cultivation.     

Late effects from particulate radiations in primate and rabbit tissues.

Optic tissues in groups of New Zealand white rabbits were irradiated locally at different stages throughout the median life span of the species with a single dose (9 Gy) of 425 MeV/amu Ne ions (LET infinity approximately 30 keV/micrometer) and then inspected routinely for the progression of radiation cataracts. The level of early cataracts was found to be highest in the youngest group of animals irradiated (8 weeks old), but both the onset of late cataracts and loss of vision occurred earlier when animals were irradiated during the second half of the median life span. This age response can have serious implications in terms of space radiation hazards to man. Rhesus monkeys that had been subjected to whole-body skin irradiation (2.8 and 5.6 Gy) by 32 MeV protons (range in tissue approximately 1 cm) some twenty years previously were analysed for radiation damage by the propagation of skin fibroblasts in primary cultures. Such propagation from skin biopsies in MEM-alpha medium (serial cultivation) or in supplemented Ham's F-10 medium (cultivation without dilution) revealed late damage in the stem (precursor) cells of the skins of the animals. The proton fluxes employed in this experiment are representative of those occurring in major solar flares.     

The "C.E.B.A.S. MINI-MODULE": a self-sustaining closed aquatic ecosystem for spaceflight experimentation.

The C.E.B.A.S. MINI-MODULE is the miniaturized space flight version of the Closed Equilibrated Biological Aquatic System (C.E.B.A.S.). It fits into a large middeck locker tray and is scheduled to be flown in the STS 85 and in the NEUROLAB missions. Its volume is about 9 liters and it consists of two animal tanks, a plant cultivator, and a bacteria filter in a monolithic design. An external sensor unit is connected to a data acquisition/control unit. The system integrates its own biological life support. The CO2 exhaled by the consumers (fishes, snails, microorganisms) is assimilated by water plants (Ceratophyllum demersum) which provide them with oxygen. The products of biomass degradation and excretion (mainly ammonia ions) are converted by bacteria into nitrite and nitrate. The latter is taken up by the plants as a nitrogen source together with other ions like phosphate. The plants convert light energy into chemical energy and their illumination is regulated via the oxygen concentration in the water by the control unit. In ground laboratory tests the system exhibited biological stability up to three month. The buffer capacity of the biological filter system is high enough to eliminate the degradation products of about one half of the dead animal biomass as shown in a "crash test". A test series using the laboratory model of the flight hardware demonstrated the biological stability and technical reliability with mission-identical loading and test duration. A comprehensive biological research program is established for the C.E.B.A.S. MINI-MODULE in which five German and three U.S.-American universities as well as the Russian Academy of Sciences are involved.     

Basic design concept of Closed Ecology Experiment Facilities.

In order to study the relationship between the physiological metabolism of living things and the environmental factors such as the atmospheric contents and so on within the closed ecosystem, Closed Ecology Experiment Facilities (CEEF) were designed and now under construction based on the following concepts: (1) Individual sealed chambers (called modules) for the plant cultivation, animal breeding, human habitation and microbial waste treatment are to be constructed independently to be able to study the metabolic effects of each living thing on the environmental factors within closed ecosystem. (2) A chamber for the microbial waste treatment are to be replaced with two systems; wet oxidation reactors and chemical nitrogen fixation reactors. (3) Atmospheric control systems are to be independently attached to each module for stabilizing atmospheric contents in each module. (4) Any construction materials having the possibility to absorb oxygen and carbon dioxide are to be prohibited to use in each module for sustaining material balance. (5) Facilities have to be developed so that the closed plant and animal experiments can be done independently, as well as integrated experiments with plants and animals through exchanging foods, carbon dioxide, oxygen, condensed water and nutrient solution.     

Light intensity and production parameters of phytocenoses cultivated on soil-like substrate under controlled [correction of controled] environment conditions.

To increase the degree of closure of biological life support systems of a new generation, we used vermicomposting to involve inedible phytomass in the intra-system mass exchange. The resulting product was a soil-like substrate, which was quite suitable for growing plants (Manukovsky et al. 1996, 1997). However, the soil like substrate can be regarded as a candidate for inclusion in a system only after a comprehensive examination of its physical, chemical, and other characteristics. An important criterion is the ability of the soil-like substrate to supply the necessary mineral elements to the photosynthesizing component under the chosen cultivation conditions. Thus, the purpose of this work was to study the feasibility of enhancing the production activity of wheat and radish crops by varying the intensity of photosynthetically active radiation, without decreasing the harvest index. The increase of light intensity from 920 to 1150 micromoles m-2 s-1 decreased the intensity of apparent photosynthesis of the wheat crops and slightly increased the apparent photosynthesis of the radish crops The maximum total and grain productivity (kg/m2) of the wheat crops was attained at the irradiance of 920 micromoles m-2 s-1. Light intensity of 1150 micromoles m-2 s-1 decreased the productivity of wheat plants and had no significant effect on the productivity of the radish crops (kg/m2) as compared to 920 micromoles m-2 s-1. The qualitative and quantitative composition of microflora of the watering solution and substrate was determined by the condition of plants, developmental phase and light intensity. By the end of wheat growth under 1150 micromoles m-2 s-1 the numbers of bacteria of the coliform family and phytopathogenic bacteria in the watering solution and substrate were an order of magnitude larger than under other illumination conditions. The obtained data suggest that the cultivation of plants in a life support system on soil-like substrate from composts has a number of advantages over the cultivation on neutral substrates, which require continual replenishment of the plant nutrient solution from the system's store to complement the macro- and micro-elements. Yet, a number of problems arise, including those related to the controlling of the production activity of the plants by the intensity of photosynthetically active radiation. It is essential to understand why the intensity of production processes is limited at higher irradiation levels and to overcome the factors responsible for this, so that the soil-like substrate could have an even better chance in the competition for the best plant cultivation technology to be used in biological life support systems.