Evaluation of Cyanothece sp. ATCC 51142 as a candidate for inclusion in a CELSS.

Controlled ecological life support systems (CELSS) have been proposed to make long-duration manned space flights more cost-effective. Higher plants will presumably provide food and a breathable atmosphere for the crew. It has been suggested that imbalances between the CO2/O2 gas exchange ratios of the heterotrophic and autotrophic components of the system will inevitably lead to an unstable system, and the loss of O2 from the atmosphere. Ratio imbalances may be corrected by including a second autotroph with an appropriate CO2/O2 gas exchange ratio. Cyanothece sp. ATCC 51142 is a large unicellular N2-fixing cyanobacterium, exhibiting high growth rates under diverse physiological conditions. A rat-feeding study showed the biomass to be edible. Furthermore, it may have a CO2/O2 gas exchange ratio that theoretically can compensate for ratio imbalances. It is suggested that Cyanothece spp. could fulfill several roles in a CELSS: supplementing atmosphere recycling, generating fixed N from the air, providing a balanced protein supplement, and protecting a CELSS in case of catastrophic crop failure.     

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.     

Development of autonomous control in a closed microbial bioreactor.

Space-based life support systems which include ecological components will rely on sophisticated hardware and software to monitor and control key system parameters. Autonomous closed artificial ecosystems are useful for research in numerous fields. We are developing a bioreactor designed to study both microbe-environment interactions and autonomous control systems. Currently we are investigating N-cycling and N-mass balance in closed microbial systems. The design features of the system involve real-time monitoring of physical parameters (e.g. temperature, light), growth solution composition (e.g. pH, NOx, CO2), cell density and the status of important hardware components. Control of key system parameters is achieved by incorporation of artificial intelligence software tools that permit autonomous decision-making by the instrument. These developments provide a valuable research tool for terrestrial microbial ecology, as well as a testbed for implementation of artificial intelligence concepts. Autonomous instrumentation will be necessary for robust operation of space-based life support systems, and for use on robotic spacecraft. Sample data acquired from the system, important features of software components, and potential applications for terrestrial and space research will be presented.     

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.     

Biosphere 2 Test Module: a ground-based sunlight-driven prototype of a closed ecological life support system.

Constructed in 1986, the Biosphere 2 Test Module has been used since the end of that year for closed ecological systems experiments. It is the largest closed ecological facility ever built, with a sealed variable volume of some 480 cubic meters. It is built with a skin of steel spaceframes with double-laminated glass panels admitting about 65 percent Photosynthetically Active Radiation (PAR). The floor is of welded steel and there is an underground atmospheric connection via an air duct to a variable volume chamber ("lung") permitting expansion and contraction of the Test Module's air volume caused by changes in temperature and barometric pressure, which causes a slight positive pressure from inside the closed system to the outside thereby insuring that the very small leakage rate is outward. Several series of closed ecological system investigations have been carried out in this facility. One series of experiments investigated the dynamics of higher plants and associated soils with the atmosphere under varying light and temperature conditions. Another series of experiments included one human in the closed system for three, five and twenty-one days. During these experiments the Test Module had subsystems which completely recycled its water and atmosphere; all the human dietary needs were produced within the facility, and all wastes were recycled using a marsh plant/microbe system. Other experiments have examined the capability of individual component systems used, such as the soil bed reactors, to eliminate experimentally introduced trace gases. Analytic systems developed for these experiments include continuous monitors of eleven atmospheric gases in addition to the complete gas chromatography mass spectrometry (GCMS) examinations of potable, waste system and irrigation water quality.     

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.     

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.     

Impaired growth of plants cultivated in a closed system: possible reasons.

Plants in experiments on "man-higher plants" closed ecosystem (CES) have been demonstrated to have inhibited growth and reduced productivity due to three basic factors: prolonged usage of a permanent nutrient solution introduction into the nutrient medium of intra-system gray water, and closure of the system. Gray water was detrimental to plants the longer the nutrient solution was used. However, higher plant growth was mostly affected by the gaseous composition of the CES atmosphere, through accumulation of volatile substances.     

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen.

Early Earth and early Mars were similar enough such that past geochemical and climatic conditions on Mars may have also been favorable for the origin of life. However, one of the most striking differences between the two planets was the low partial pressure of dinitrogen (pN2) on early Mars (18 mb). On Earth, nitrogen is a key biological element and in many ecosystems the low availability of fixed nitrogen compounds is the main factor limiting growth. Biological fixation of dinitrogen on Earth is a crucial source of fixed nitrogen. Could the low availability of dinitrogen in the primordial Martian atmosphere have prevented the existence, or evolution of Martian microbiota? Azotobacter vinelandii and Azomonas agilis were grown in nitrogen free synthetic medium under various partial pressures of dinitrogen ranging from 780-0 mb (total atmosphere=1 bar). Below 400 mb the biomass, cell number, and growth rate decreased with decreasing pN2. Both microorganisms were capable of growth at a pN2 as low as 5 mb, but no growth was observed at a pN2 < or = 1 mb. The data appear to indicate that biological nitrogen fixation could have occurred on primordial Mars (pN2=18 mb) making it possible for a biotic system to have played a role in the Martian nitrogen cycle. It is possible that nitrogen may have played a key role in the early evolution of life on Mars, and that later a lack of available nitrogen on that planet (currently, pN2=0.2 mb) may have been involved in its subsequent extinction.     

利用WACCM-3模式对平流层动力、热力场及微量化学成分季节变化的数值模拟研究

利用美国NCAR最新的化学-气候耦合模式WACCM-3对平流层风场、温度场以及平流层臭氧等多种微量气体成分(O3, CH4, N2O, H2O, HCl, HNO3)的季节变化进行了数值模拟, 并使用ECMWF再分析资料与美国UARS卫星 搭载的HALOE, MLS, CLAES等探测器的观测资料, 对模式输出的动力、热力及化学成分浓度的气候平均值进行了验证. 结果表明, 在气候平均海表温度值驱动下, WACCM-3模式能够很好地再现ECMWF资料中平流层纬向平均风场与温度场的季节变化. 模拟结果中平流层化学成分的经向-垂直分布及其季节变化与卫星观测结果基本一致. 模式的动力、热力场在极地平流层以及热带对流层顶等区域存在一定的偏差. 这些偏差对于微量气体成分分布 的模拟具有一定影响, 特别是南半球冬(7月)、春(10月)季节南极平流层低层极夜 急流偏强, 造成极地地区附近的输送障碍增强, 从而导致CH4, N2O, H2O浓度比观测偏低. 此外, WACCM-3缺少热带平流层风场的准两年振荡(QBO) 机制, 这对于热带平流层东风急流以及低纬度平流层O3, CH4, N2O, H2O等成分经向输送的模拟结果也有一定影响.      

Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems.

Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. We report experiments in which cultures of the algae Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments we tested hydrolyzed waste biomass from these same algae to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.     

Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor.

Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (1 g). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of E11.5 limb buds was cultured with and without encapsulation in alginate prior to culturing in the bioreactor. Encapsulated limbs grown in the bioreactor did not fuse together, but developed only the more proximal elements while limbs grown in culture dishes formed proximal and distal elements. Alginate encapsulation may have reduced oxygenation to the progress zone of the developing limb bud resulting in lack of development of the more distal elements. These results show that the bioreactor supports growth and differentiation of skeletal elements in entire E13 limb buds, and that a method to culture younger limb buds without fusing together needs to be developed if any morphometric analysis is to be performed.     

Inhibition of denitrification by ultraviolet radiation.

It has been shown that UV-A (lambda=320-400 nm) and UV-B (lambda=280-320 nm) inhibit photosynthesis, nitrogen fixation and nitrification. The purpose of this study was to determine the effects, if any, on denitrification in a microbial community inhabiting the intertidal. The community studied is the microbial mat consisting primarily of Lyngbya that inhabits the Pacific marine intertidal, Baja California, Mexico. Rates of denitrification were determined using the acetylene blockage technique. Pseudomonas fluorescens (ATCC #17400) was used as a control organism, and treated similarly to the mat samples. Samples were incubated either beneath a PAR transparent, UV opaque screen (OP3), or a mylar screen to block UV-B, or a UV transparent screen (UVT) for 2 to 3 hours. Sets of samples were also treated with nitrapyrin to inhibit nitrification, or DCMU to inhibit photosynthesis and treated similarly. Denitrification rates were greater in the UV protected samples than in the UV exposed samples the mat samples as well as for the Ps fluorescens cultures. Killed controls exhibited no activity. In the DCMU and nitrapyrin treated samples denitrification rates were the same as in the untreated samples. These data indicate that denitrification is directly inhibited by UV radiation.     

Evaluation of an anaerobic digestion system for processing CELSS crop residues for resource recovery.

Three bioreactors, connected in series, were used to process CELSS potato residues for recovery of resources. The first stage was an anaerobic digestor (8 L working volume; cow rumen contents inoculum; fed-batch; 8 day retention time; feed rate 25 gdw day-1) that converted 33% of feed (dry weight loss) to CO2 and "volatile fatty acids" (vfa, 83:8:8 mmolar ratio acetic:propionic:butyric). High nitrate-N in the potato residue feed was absent in the anaerobic effluent, with a high portion converted to NH4(+)-N and the remainder unaccounted and probably lost to denitrification and NH4+ volatilization. Liquid anaerobic effluent was fed to an aerobic, yeast biomass production vessel (2 L volume; Candida ingens inoculum; batch [pellicle] growth; 2 day retention time) where the VFAs and some NH4(+)-N were converted into yeast biomass. Yeast yields accounted for up to 8% of potato residue fed into the anaerobic bioreactor. The third bioreactor (0.5 L liquid working volume; commercial nitrifier inoculum; packed-bed biofilm; continuous yeast effluent feed; recirculating; constant volume; 23 day hydraulic retention time) was used to convert successfully the remaining NH4(+)-N into nitrate-N (preferred form of N for CELSS crop production) and to remove the remaining degradable soluble organic carbon. Effluents from the last two stages were used for partial replenishment of minerals for hydroponic potato production.     

太阳表面水平运动驱动的磁圈与开放磁场重联的模拟研究

太阳大气的诸多观测事件(如耀斑、喷流等)均被归因于磁重联产生的能量转换. 近年来, 关于太阳风起源, 有研究提出了磁重联使闭合磁圈开放为太阳风供应物质的新模式. 在该模式中, 闭合磁圈被光球超米粒组织对流携带, 向超米粒边界运动, 与位于边界的开放磁场相碰撞进而发生磁重联. 该模式中磁重联的驱动及其效应是本文的研究目标. 磁流体力学(MHD)数值模拟是研究太阳大气磁重联物理过程的重要途径. 本文建立了一个二维MHD数值模型, 结合太阳大气温度和密度的分层分布, 在超米粒组织尺度上模拟了水平流动驱动的闭合磁圈与开放磁场的重联过程. 通过对模拟结果的定量分析, 认为磁重联确实能够将闭合磁圈的物质释放, 进而供应给新的开放磁结构并产生向上流动. 该结果为进一步模拟研究太阳风初始外流奠定了基础.      

A simplified closed artificial ecosystem--recycling of organic matter into wheat plants.

A simplified closed system consisting of a plant growth chamber coupled to a decomposition chamber was used to study carbon exchange dynamics. The CO2 produced via the decomposition of wheat straw was used for photosynthetic carbon uptake by wheat plants. The atmosphere of the two chambers was connected through a circuit of known flow rate. Thus, monitoring the CO2 concentrations in both compartments allowed measurement of the carbon exchange between the chambers, and estimation of the rate of respiration processes in the decomposition chamber and photosynthetic rate in the producer chamber. The objective for CELSS research was to simulate a system where a compartment producing food via photosynthesis, would be supplied by CO2 produced from respiration processes. The decomposition of biomass by the decomposer simulated both the metabolism of a crew and the result of a recycling system for inedible biomass. Concerning terrestrial ecosystems, the objective was to study organic matter decomposition in soil and other processes related to permanent grasslands.     

Assessing the feasibility of involving gaseous products resulting from physicochemical oxidation of human liquid and solid wastes in the cycling of a bio-technical life support system

The study addresses the possible ways of involving gaseous products produced by “wet” incineration of human wastes mixed with H₂O₂ in an alternating electric field in the cycling of the physical model of a bio-technical life support system (BTLSS). The resulting gas mixture contains CO₂ and O₂, which are easily involved in the cycling in the closed ecosystem, and NH₃, which is unacceptable in the atmosphere of the BTLSS. NH₃ fixation has been proposed, which is followed by nitrification and involvement of the resulting products in the mass exchange of the closed system. Experiments have been performed to show that plants can be grown in the atmosphere resulting from the closing of the gas loop that includes a physicochemical installation and a growth chamber with plants representing the phototrophic compartment of the BTLSS. The results of the study suggest the conclusion that the proposed method of organic waste oxidation can be a useful tool in creating a physical model of a closed-loop integrated BTLSS.     

人为活动对高平流层,中间层大气的影响

利用一维光化、辐射耦合模式研究了人为活动所排放出的废弃物ＣＨ４、Ｎ２Ｏ和ＣＯ２含量增加对高平流层，中间层大气的影响。结果指出，对流层中这些气体增加一倍将导致上中层大气水汽、Ｏ３和温度的明显变化。ＣＯ２含量增加将使上中层温度降低约１０Ｋ；而ＣＨ４含量的加倍不仅使中层顶附近水汽含量增加５０％以上，也使其温度有更明显的下降，这将更有利于於光云的形成。     

Earth applications of closed ecological systems: relevance to the development of sustainability in our global biosphere.

The parallels between the challenges facing bioregenerative life support in artificial closed ecological systems and those in our global biosphere are striking. At the scale of the current global technosphere and expanding human population, it is increasingly obvious that the biosphere can no longer safely buffer and absorb technogenic and anthropogenic pollutants. The loss of biodiversity, reliance on non-renewable natural resources, and conversion of once wild ecosystems for human use with attendant desertification/soil erosion, has led to a shift of consciousness and the widespread call for sustainability of human activities. For researchers working on bioregenerative life support in closed systems, the small volumes and faster cycling times than in the Earth's biosphere make it starkly clear that systems must be designed to ensure renewal of water and atmosphere, nutrient recycling, production of healthy food, and safe environmental methods of maintaining technical systems. The development of technical systems that can be fully integrated and supportive of living systems is a harbinger of new perspectives as well as technologies in the global environment. In addition, closed system bioregenerative life support offers opportunities for public education and consciousness changing of how to live with our global biosphere.     

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass.

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estunate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.