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.     

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.     

Direct utilization of human liquid wastes by plants in a closed ecosystem.

Model experiments in phytotrons have shown that urea is able to cover 70% of the demand in nitrogen of the conveyer cultivated wheat. At the same time wheat plants can directly utilize human liquid wastes. In this article by human liquid wastes the authors mean human urine only. In a long-term experiment on "man-higher plants" system with two crewmen, plants covered 63 m2, with wheat planted to--39.6 m2. For 103 days, complete human urine (total amount--210.7 l) was supplied into the nutrient solution for wheat. In a month and a half NaCl supply into the nutrient solution stabilized at 0.9-1.65 g/l. This salination had no marked effect on wheat production. The experiment revealed the realistic feasibility to directly involve liquid wastes into the biological turnover of the life support system. The closure of the system, in terms of water, increased by 15.7% and the supply of nutrients for wheat plants into the system was decreased.     

A biological method of including mineralized human liquid and solid wastes into the mass exchange of bio-technical life support systems

The main obstacle to using mineralized human solid and liquid wastes as a source of mineral elements for plants cultivated in bio-technical life support systems (BLSS) is that they contain NaCl. The purpose of this study is to determine whether mineralized human wastes can be used to prepare the nutrient solution for long-duration conveyor cultivation of uneven-aged wheat and Salicornia europaea L. plant community. Human solid and liquid wastes were mineralized by the method of “wet incineration” developed by Yu. Kudenko. They served as a basis for preparing the solutions that were used for conveyor-type cultivation of wheat community represented by 5 age groups, planted with a time interval of 14 days. Wheat was cultivated hydroponically on expanded clay particles. To reduce salt content of the nutrient solution, every two weeks, after wheat was harvested, 12 L of solution was removed from the wheat irrigation tank and used for Salicornia europaea cultivation in water culture in a conveyor mode. The Salicornia community was represented by 2 age groups, planted with a time interval of 14 days. As some portion of the nutrient solution used for wheat cultivation was regularly removed, sodium concentration in the wheat irrigation solution did not exceed 400 mg/L, and mineral elements contained in the removed portion were used for Salicornia cultivation. The experiment lasted 4 months. The total wheat biomass productivity averaged 30.1 g · m⁻² · day⁻¹, and the harvest index amounted to 36.8%. The average productivity of Salicornia edible biomass on a dry weight basis was 39.3 g · m⁻² · day⁻¹, and its aboveground mass contained at least 20% of NaCl. Thus, the proposed technology of cultivation of wheat and halophyte plant community enables using mineralized human wastes as a basis for preparing nutrient solutions and including NaCl in the mass exchange of the BLSS; moreover, humans are supplied with additional amounts of leafy vegetables.     

Biological and physiochemical methods for utilization of plant wastes and human exometabolites for increasing internal cycling and closure of life support systems.

Wheat was cultivated on soil-like substrate (SLS) produced by the action of worms and microflora from the inedible biomass of wheat. After the growth of the wheat crop, the inedible biomass was restored in SLS and exposed to decomposition ("biological" combustion) and its mineral compounds were assimilated by plants. Grain was returned to the SLS in the amount equivalent to human solid waste produced by consumption of the grain. Human wastes (urine and feces) after physicochemical processing turned into mineralized form (mineralized urine and mineralized feces) and entered the plants' nutrient solution amounts equal to average daily production. Periodically (once every 60-70 days) the nutrient solution was partly (up to 50%) desalinated by electrodialysis. Due to this NaCl concentration in the nutrient solution was sustained at a fixed level of about 0.26%. The salt concentrate obtained could be used in the human nutrition through NaCl extraction and the residuary elements were returned through the mineralized human liquid wastes into matter turnover. The control wheat cultivation was carried out on peat with use of the Knop nutrient solution. Serial cultivation of several wheat vegetations within 280 days was conducted during the experiment. Grain output varied and yield/harvest depended, in large part, upon the amount of inedible biomass returned to SLS and the speed of its decomposition. After achieving a stationary regime, (when the quantity of wheat inedible biomass utilized during vegetation in SLS is equal to the quantity of biomass introduced into SLS before vegetation) grain harvest in comparison with the control was at most 30% less, and in some cases was comparable to the control harvest values. The investigations carried out on the wheat example demonstrated in principle the possibility of long-term functioning of the LSS photosynthesizing link based on optimizations of biological and physicochemical methods of utilization of the human and plants wastes. The possibilities for the use of these technologies for the creation integrated biological-physicochemical LSS with high closure degree of internal matter turnover are discussed in this paper.     

A conceptual configuration of the lunar base bioregenerative life support system including soil-like substrate for growing plants

The paper presents a conceptual configuration of the lunar base bioregenerative life support system (LBLSS), including soil-like substrate (SLS) for growing plants. SLS makes it possible to combine the processes of plant growth and the utilization of plant waste. Plants are to be grown on SLS on the basis of 20 kg of dry SLS mass or 100 kg of wet SLS mass per square meter. The substrate is to be delivered to the base ready-made as part of the plant growth subsystem. Food for the crew was provided by prestored stock 24% and by plant growing system 76%. Total dry weight of the food is 631 g per day (2800 kcal/day) for one crew member (CM). The list of candidate plants to be grown under lunar BLSS conditions included 14 species: wheat, rice, soybean, peanuts, sweet pepper, carrots, tomatoes, coriander, cole, lettuce, radish, squash, onion and garlic. From the prestored stock the crew consumed canned fish, iodinated salt, sugar, beef sauce and seafood sauce. Our calculations show that to provide one CM with plant food requires the area of 47.5 m². The balance of substance is achieved by the removal dehydrated urine 59 g, feces 31 g, food waste 50 g, SLS 134 g, and also waters 86 g from system and introduction food 236 g, liquid potassium soap 4 g and mineral salts 120 g into system daily. To reduce system setup time the first plants could be sowed and germinated to a certain age on the Earth.     

Use of halophytic plants for recycling NaCl in human liquid waste in a bioregenerative life support system

The purpose of this work was to develop technology for recycling NaCl containing in human liquid waste as intrasystem matter in a bioregenerative life support system (BLSS). The circulation of Na⁺ and Cl⁻ excreted in urine is achieved by inclusion of halophytes, i.e. plants that naturally inhabit salt-rich soils and accumulate NaCl in their organs. A model of Na⁺ and Cl⁻ recycling in a BLSS was designed, based on the NaCl turnover in the human–urine–nutrient solution–halophytic plant–human cycle. The study consisted of (i) selecting a halophyte suitable for inclusion in a BLSS, and (ii) determining growth conditions supporting maximal Na⁺ and Cl⁻ accumulation in the shoots of the halophyte growing in a nutrient solution simulating mineralized urine. For the selected halophytic plant, Salicornia europaea, growth rate under optimal conditions, biomass production and quantities of Na⁺ and Cl⁻ absorbed were determined. Characteristics of a plant production conveyor consisting of S.europaea at various ages, and allowing continuity of Na⁺ and Cl⁻ turnover, were estimated. It was shown that closure of the NaCl cycle in a BLSS can be attained if the daily ration of fresh Salicornia biomass for a BLSS inhabitant is approximately 360 g.     

Productivity and food value of Amaranthus cruentus under non-lethal salt stress.

Stress effects from the accumulation of metal salts may pose a problem for plants in closed biological systems such as spacecraft. This work examined the effects of salinity on growth, photosynthesis and carbon allocation in the crop plant, Amaranthus. Plants were germinated and grown in modified Hoagland's solution with NaCl concentrations of 0 to 1.0%. Plants received salt treatments at various times in development to assess effects on particular life history phases. For Amaranthus cruentus, germination, vegetative growth, flowering, seed development and yield were normal at salinities from 0 to 0.2%. Inhibition of these phases increased from 0.2 to 0.4% salinity and was total above 0.5%. 1.0% salinity was lethal to all developmental phases. Onset of growth phases were not affected by salinity. Plants could not be adapted by gradually increasing salinity over days or weeks. Water uptake increased, while photosynthetic CO2 uptake decreased with increasing salinity on a dry weight basis during vegetative growth. Respiration was not affected by salinity. After flowering, respiration and photosynthesis decreased markedly, such that 1.0% NaCl inhibited photosynthesis completely. Protein levels were unchanged with increasing salinity. Leaf starch levels were lower at salinities of 0.5% and above, while stem starch levels were not affected by these salinities. The evidence supports salt inhibition arising from changes in primary biochemical processes rather than from effects on water relations. While not addressing the toxic effects of specific ions, it suggests that moderate salinity per se need not be a problem in space systems.     

Use of human wastes oxidized to different degrees in cultivation of higher plants on the soil-like substrate intended for closed ecosystems

To close mass exchange loops in bioregenerative life support systems more efficiently, researchers of the Institute of Biophysics SB RAS (Krasnoyarsk, Russia) have developed a procedure of wet combustion of human wastes and inedible parts of plants using H₂O₂ in alternating electromagnetic field. Human wastes pretreated in this way can be used as nutrient solutions to grow plants in the phototrophic unit of the LSS. The purpose of this study was to explore the possibilities of using human wastes oxidized to different degrees to grow plants cultivated on the soil-like substrate (SLS). The treated human wastes were analyzed to test their sterility. Then we investigated the effects produced by human wastes oxidized to different degrees on growth and development of wheat plants and on the composition of microflora in the SLS. The irrigation solution contained water, substances extracted from the substrate, and certain amounts of the mineralized human wastes. The experiments showed that the human wastes oxidized using reduced amounts of 30% H₂O₂: 1 ml/g of feces and 0.25 ml/ml of urine were still sterile. The experiments with wheat plants grown on the SLS and irrigated by the solution containing treated human wastes in the amount simulating 1/6 of the daily diet of a human showed that the degree of oxidation of human wastes did not significantly affect plant productivity. On the other hand, the composition of the microbiota of irrigation solutions was affected by the oxidation level of the added metabolites. In the solutions supplemented with partially oxidized metabolites yeast-like microscopic fungi were 20 times more abundant than in the solutions containing fully oxidized metabolites. Moreover, in the solutions containing incompletely oxidized human wastes the amounts of phytopathogenic bacteria and denitrifying microorganisms were larger. Thus, insufficiently oxidized sterile human wastes added to the irrigation solutions significantly affect the composition of the microbiological component of these solutions, which can ultimately unbalance the system as a whole.     

载人深空探测活动中的尿液处理回收技术分析

生物再生生命保障系统(Bioregenerative Life Support System,BLSS)是人类进行深空探测活动,实现长期载人空间飞行必需的关键技术,对于太空的探索开发具有重要意义。在BLSS系统内,航天员尿液废水的处理回收是非常重要的一部分。将尿液中所含有的大量的水分和丰富的营养物质回收用于系统内植物生长所需营养液的配制,既可以保证植物的正常生长,也有助于实现系统内物质的循环利用进而提高BLSS的闭合度。尿液中所含的大量盐分会威胁植物生长,所以需通过一定的技术手段处理尿液废水并回收其中的水分和营养。为了探索适用于BLSS中的尿液处理回收技术,首先分析了几种面向空间站应用的尿液处理技术,如蒸馏技术等;然后基于回收营养物质的需求,分析了面向民用的、发展较为成熟的尿液处理回收技术,如离子交换吸附技术、氨气吹脱技术和鸟粪石沉淀技术,并讨论了这些尿液处理回收技术在BLSS中的应用前景。最后基于BLSS的实际需求,提出了有望用于BLSS中的尿液处理回收技术流程。     

Plants--rhizospheric organisms interaction in a manmade system with and without biogenous element limitation.

The effect has been studied of inoculation of seeds of wheat with two species of rhizospheric microorganisms--Pseudomonas fluorescens and Pseudomonas putida--on young plant growth with complete and with nitrogen deficit mineral nutrition. With complete mineral medium, plants grown from seeds inoculated with bacteria of Pseudomonas genus (experiment plants) have been found to have better growth over plants not inoculated with these bacteria (control plants). The experiment plants had increased transpiration and their biomass had higher organic nitrogen content. With nitrogen deficit medium, the plants inoculated with bacteria and those without them, have not revealed changes in growth. Neither case demonstrated competition of microorganisms with plants for nitrogen sources.     

Effects of salts and temperatures on post-irradiation growth of penicillium exposed to ultraviolet

The growth of Penicillium notatum colonies after UV irradiation of dried mycelium or spores was studied in relation to post-irradiation temperature and salt environment. Dried mycelium and spores behaved differently with respect to sensitivity to temperature, salts and UV, especially the latter. Threshold inhibitory doses for spores were modified markedly either at 4°C or in magnesium and calcium chlorides. It is suggested that these temperature and salt effects are related to prevention of photochemical membrane damage.     

Effects of mineral nutrition conditions on heat tolerance of chufa (Сyperus esculentus L.) plant communities to super optimal air temperatures in the BTLSS

The use of mineralized human wastes as a basis for nutrient solutions will increase the degree of material closure of bio-technical human life support systems. As stress tolerance of plants is determined, among other factors, by the conditions under which they have been grown before exposure to a stressor, the purpose of the study is to investigate the level of tolerance of chufa (Cyperus esculentus L.) plant communities grown in solutions based on mineralized human wastes to a damaging air temperature, 45 °C. Experiments were performed with 30-day-old chufa plant communities grown hydroponically, on expanded clay aggregate, under artificial light, at 690 μmol m⁻² s⁻¹ PAR and at a temperature of 25 °C. Plants were grown in Knop’s solution and solutions based on human wastes mineralized according to Yu.A. Kudenko’s method, which contained nitrogen either as ammonium and urea or as nitrates. The heat shock treatment lasted 20 h at 690 and 1150 μmol m⁻² s⁻¹ PAR. Chufa heat tolerance was evaluated based on parameters of CO₂ gas exchange, the state of its photosynthetic apparatus (PSA), and intensity of peroxidation of leaf lipids. Chufa plants grown in the solutions based on mineralized human wastes that contained ammonium and urea had lower heat tolerance than plants grown in standard mineral solutions. Heat tolerance of the plants grown in the solutions based on mineralized human wastes that mainly contained nitrate nitrogen was insignificantly different from the heat tolerance of the plants grown in standard mineral solutions. A PAR intensity increase from 690 μmol m⁻² s⁻¹ to 1150 μmol m⁻² s⁻¹ enhanced heat tolerance of chufa plant communities, irrespective of the conditions of mineral nutrition under which they had been grown.     

Gravity and the membrane-solution interface: theoretical investigations.

The theory of concentration and potential variations at interfaces is applied to the membrane-solution interface to calculate density variations. The theory is modified to take care of the finite ion volumes in electrolytes. Our model is a phospholipid membrane with a surface charge density of -4.824*10(-6)(As/cm2) in contact with solutions of KCl, NaCl, CaCl2, and mixtures. Maximal density variations of about 4*10(-2)(G/cm3) were found in surface layers between the membrane and the solutions. The extension of the layers is in the range of 1 to 6 nm.     

Increased BLSS closure using mineralized human waste in plant cultivation on a neutral substrate

The purpose of this work was to study the full-scale potential use of human mineralized waste (feces and urine) as a source of mineral elements for plant cultivation in a biological life support system (BLSS). Plants that are potential candidates for a photosynthesizing link were grown on a neutral solution containing human mineralized waste. Spring wheat Triticum aestivum L., peas Pisum sativum L. Ambrosia cultivar and leaf lettuce Lactuca sativa L., Vitaminny variety, were used. The plants were grown hydroponically on expanded clay aggregates in a vegetation chamber in constant environmental conditions. During plant growth, a determined amount of human mineralized waste was added daily to the nutrient solution. The nutrient solution remained unchanged throughout the vegetation period. Estimated plant requirements for macro-elements were based on a total biological productivity of 0.04 kg day⁻¹ m⁻². As the plant requirements for potassium exceeded the potassium content of human waste, a water extract of wheat straw containing the required amount of potassium was added to the nutrient solution. The Knop’s solution was used in the control experiments.     

氯盐对碱激发矿渣净浆强度影响试验

通过对掺加氯化钠(NaCl)、氯化钙(CaCl₂)的碱激发矿渣(KC)净浆的无侧限抗压强度试验,研究两种氯盐对KC净浆强度的影响规律,利用X射线衍射(XRD)、热重分析(TG-DTG)、液相离子测定、水化程度测定等测试手段,分析两种氯盐对KC净浆强度的作用机理.结果表明,随着NaCl掺量增加,KC净浆抗压强度提高;随着CaCl₂掺量增加,KC净浆抗压强度基本不变.掺加NaCl与CaCl₂的KC净浆中均生成含氯水化物水化氯铝酸钙(Fs,Friedel's salt),但Fs的生成对KC净浆强度无影响作用.掺加NaCl的KC净浆中氢氧化钠(NaOH)的生成是导致其强度增强的主要原因,混合物中NaOH的存在提高了KC净浆的液相碱度,促进矿渣水化的进一步发生,进而生成更多的水化硅酸钙(CSH)使KC净浆强度得到提高.      

Analysis of mutant quantity and quality in human-hamster hybrid AL and AL-179 cells exposed to 137Cs-gamma or HZE-Fe ions.

We measured the number of mutants and the kinds of mutations induced by 137Cs-gamma and by HZE-Fe (56Fe [600 MeV/amu, LET = 190 KeV/micrometer) in standard AL human hamster hybrid cells and in a new variant hybrid, AL-179. We found that HZE-Fe was more mutagenic than 137Cs-gamma per unit dose (about 1.6 fold), but was slightly less mutagenic per mean lethal dose, DO, at both the S1 and hprt- loci of AL cells. On the other hand, HZE-Fe induced about nine fold more complex S1- mutants than 137Cs-gamma rays, 28% vs 3%. 137Cs-gamma rays induced about twice as many S1- mutants and hprt-mutants in AL-179 as in AL cells, and about nine times more of the former were complex, and potentially unstable kinds of mutations.     

Clinostating effects on biochemical characteristics and productivity of healthy and virus-infected wheat plants of dwarf Apogee variety.

The effects of clinostating on physiological processes and biochemical characteristics of wheat plants (Triticum aestivum L.) both healthy and infected by the wheat streak mosaic virus (WSMV) were studied. In six experiments, each lasting over 30 days, healthy and infected plants of the dwarf Apogee variety were grown under conditions of continuous horizontal and vertical clinostating with 2 rpm at 21 +/- 2 degrees C and 6000 1x (the optimal moisture of a substrate being maintained). The control variants (healthy and infected) were simultaneously grown under the same conditions of temperature and illumination in stationary containers and in open pots. During the experiment, visual observations were carried out over the state of tested plants. After completing the experiment, biometric indices, pigment, carbohydrate and dry matter contents were determined in all the plants. It was shown that clinostating sharply reduced the reproductive function of healthy plants and considerably affected their biomass (productivity) and concentration of chlorophylls and sugars. The viral infection resulted in further reduction of these characteristics. In control variants the viral effect was more significant. We speculate that clinostating reduced the rate of reproduction and spread of the virus.     

Composition and physical properties of starch in microgravity-grown plants.

The effect of spaceflight on starch development in soybean (Glycine max L., BRIC-03) and potato (Solanum tuberosum, Astroculture-05) was compared with ground controls by biophysical and biochemical measurements. Starch grains from plants from both flights were on average 20-50% smaller in diameter than ground controls. The ratio delta X/delta rho (delta X --difference of magnetic susceptibilities, delta rho--difference of densities between starch and water) of starch grains was ca. 15% and 4% higher for space-grown soybean cotyledons and potato tubers, respectively, than in corresponding ground controls. Since the densities of particles were similar for all samples (1.36 to 1.38 g/cm3), the observed difference in delta X/delta rho was due to different magnetic susceptibilities and indicates modified composition of starch grains. In starch preparations from soybean cotyledons (BRIC-03) subjected to controlled enzymatic degradation with alpha-amylase for 24 hours, 77 +/- 6% of the starch from the flight cotyledons was degraded compared to 58 +/- 12% in ground controls. The amylose content in starch was also higher in space-grown tissues. The good correlation between the amylose content and delta X/delta rho suggests, that the magnetic susceptibility of starch grains is related to their amylose content. Since the seedlings from the BRIC-03 experiment showed elevated post-flight ethylene levels, material from another flight experiment (GENEX) which had normal levels of ethylene was examined and showed no difference to ground controls in size distribution, density, delta X/delta rho and amylose content. Therefore the role of ethylene appears to be more important for changes in starch metabolism than microgravity.     

C.E.B.A.S. MINI MODULE: test results of an artificial (man-made) aquatic ecosystem.

The original Closed Equilibrated Biological Aquatic System (C.E.B.A.S.) is a long-term multi-generation research facility for experiments with aquatic animals and plants in a space station the development of which is surrounded by a large international scientific program. In addition, a miniaturized laboratory prototype, the C.E.B.A.S. MINI MODULE, with a total volume of about 10-12 liters for a Spacelab middeck locker was developed and a first version was tested successfully for two weeks with a population of fishes (Xiphophorus helleri) in the animal tank and a Ceratophyllum spec. in the illuminated higher plant growth chamber. The water recycling system consisted of a bacteria filter and a mechanical filter and the silastic tubing gas exchanger was separated by valves for the utilization in emergency cases only. Data were collected with the acquisition module of the original C.E.B.A.S. process control system. In addition, an optimized version was tested for 7 weeks with fishes and plants and thereafter with fish and with plants only for 2 and 1 weeks, resp.. The paper presents the relevant water parameters (e.g., pH, pressure, temperature, oxygen saturation, flow rate, ion concentrations) during the test period as well as morphological and physiological data of the enclosed animals and plants. On the basis of the given results the possible role of the C.E.B.A.S. system as a scientific tool in artificial ecosystem research and for the development of a combined animal-plant intensive aquaculture system and its utilization in bioregenerative life support is discussed.