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.     

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.     

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.     

Extraction of mineral elements from inedible wastes of biological components of a life-support system and their utilization for plant nutrition.

Two methods of extracting mineral elements from otherwise deadlock products of a life-support system are presented. We describe first optimum conditions for recovering elements by water extraction from dry wastes of plants, biomass ash, and solid human wastes after passing them through the catalytic furnace; and, second, we describe acid extracts of biogenous elements by 1N and 2N HNO3 from these products. Ways to use the extracts of elements in plant nutrition are considered in order to increase the extent to which the mineral loop of a life-support system can be closed.     

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

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

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.     

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.     

Use of lunar regolith as a substrate for plant growth.

Regenerative Life Support Systems (RLSS) will be required to regenerate air, water, and wastes, and to produce food for human consumption during long-duration missions to the Moon and Mars. It may be possible to supplement some of the materials needed for a lunar RLSS from resources on the Moon. Natural materials at the lunar surface may be used for a variety of lunar RLSS needs, including (i) soils or solid-support substrates for plant growth, (ii) sources for extraction of essential, plant-growth nutrients, (iii) substrates for microbial populations in the degradation of wastes, (iv) sources of O2 and H2, which may be used to manufacture water, (v) feed stock materials for the synthesis of useful minerals (e.g., molecular sieves), and (vi) shielding materials surrounding the outpost structure to protect humans, plants, and microorganisms from harmful radiation. Use of indigenous lunar regolith as a terrestrial-like soil for plant growth could offer a solid support substrate, buffering capacity, nutrient source/storage/retention capabilities, and should be relatively easy to maintain. The lunar regolith could, with a suitable microbial population, play a role in waste renovation; much like terrestrial waste application directly on soils. Issues associated with potentially toxic elements, pH, nutrient availability, air and fluid movement parameters, and cation exchange capacity of lunar regolith need to be addressed before lunar materials can be used effectively as soils for plant growth.     

Long-term experiments on man's stay in biological life-support system.

We describe the experimental system having maximal possible closure of material recycling in an ecosystem, including people and plants, which was carried out in a hermetically sealed experimental complex "BIOS-3", 315 m2 in volume. The system included 2 experimentators and 3 phytotrons with plants (total sowing area of 63 m2). Plants were grown with round-the-clock lamp irradiation with 130 Wm-2 PAR intensity. The plants production was food for people. Water exchange of ecosystem, as wall as gas exchange, was fully closed excluding liquids and gas samples taken for chemical analysis outside the system. The total closure of material turnover constituted 91%. Health state of the crew was estimated before, during and after the experiment. A 5-months period did not affect their health. The experiments carried out prove that the closed ecosystem of "man-plants" is a prototype of a life-support system for long-term space expeditions.     

Stoichiometric constraints and complete closure of long-term life support systems.

High closure of matter recycling is an obvious requirement for long-term life support systems (LSS). Biological species are obligate components of the LSS since physical/chemical components are not able yet to provide food for crew. However including biological species into LSS is difficult due to specific stoichiometric configuration of their inputs and outputs. Formally the problem is to estimate the ability for given set of species to provide complete closure of LSS. Two possible models of metabolism organization can be considered: rigid and flexible ones. Stoichiometric analyses showed that the rigid metabolism case is not typical and takes place with very specific requirements. The flexible metabolic model can be applied to describing wide range of systems. Some formal indications of ability to provide complete closure and stationarity of LSS state are considered in the paper. These indications establish some constraints on the form of mathematical models intended to describe artificial and natural ecological systems.     

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.     

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.     

Tolerance of wheat and lettuce plants grown on human mineralized waste to high temperature stress

The main objective of a life support system for space missions is to supply a crew with food, water and oxygen, and to eliminate their wastes. The ultimate goal is to achieve the highest degree of closure of the system using controlled processes offering a high level of reliability and flexibility. Enhancement of closure of a biological life support system (BLSS) that includes plants relies on increased regeneration of plant waste, and utilization of solid and liquid human wastes. Clearly, the robustness of a BLSS subjected to stress will be substantially determined by the robustness of the plant components of the phototrophic unit. The aim of the present work was to estimate the heat resistance of two plants (wheat and lettuce) grown on human wastes. Human exometabolites mineralized by hydrogen peroxide in an electromagnetic field were used to make a nutrient solution for the plants. We looked for a possible increase in the heat tolerance of the wheat plants using changes in photosynthetically active radiation (PAR) intensity during heat stress. At age 15 days, plants were subjected to a rise in air temperature (from 23 ± 1 °C to 44 ± 1 °С) under different PAR intensities for 4 h. The status of the photosynthetic apparatus of the plants was assessed by external СО₂ gas exchange and fluorescence measurements. The increased irradiance of the plants during the high temperature period demonstrated its protective action for both the photosynthetic apparatus of the leaves and subsequent plant growth and development. The productivity of the plants subjected to temperature changes at 250 W m⁻² of PAR did not differ from that of controls, whereas the productivity of the plants subjected to the same heat stress but in darkness was halved.     

An alternative approach to solar system exploration providing safety of human mission to Mars.

For systematic human Mars exploration, meeting crew safety requirements, it seems perspective to assemble into a spacecraft: an electrical rocket, a well-shielded long-term life support system, and a manipulator-robots operating in combined "presence effect" and "master-slave" mode. The electrical spacecraft would carry humans to the orbit of Mars, providing short distance (and low signal time delay) between operator and robot-manipulators, which are landed on the surface of the planet. Long-term hybrid biological and physical/chemical LSS could provide environment supporting human health and well being. Robot-manipulators operating in "presence effect" and "master-slave" mode exclude necessity of human landing on Martian surface decreasing the level of risk for crew. Since crewmen would not have direct contact with the Martian environment then the problem of mutual biological protection is essentially reduced. Lightweight robot-manipulators, without heavy life support systems and without the necessity of returning to the mother vessel, could be sent as scouts to different places on the planet surface, scanning the most interesting for exobiological research site. Some approximate estimations of electric spacecraft, long-term hybrid LSS, radiation protection and mission parameters are conducted and discussed.     

The biological component of the life support system for a Martian expedition.

Ground-based experiments at RF SSC-IBMP RAS (State Science Center of Russian Federation--Institute of Biomedical Problems of Russian Academia of Science) were aimed at overall studies of a human-unicellular algae-mineralization LSS (life support system) model. The system was 15 m3 in volume. It contained 45 L of algal suspension with a dry substance density of 10-12 g per liter; water volume, including the algal suspension, was 59 L. More sophisticated model systems with partial substitution of unicellular algae with higher plates (crop area of 15 m2) were tested in three experiments from 1.5 to 2 months in duration. The experiments demonstrated that LSS employing the unicellular algae play not only a macrofunction (regeneration of atmosphere and water) but also carry some other functions (purification of atmosphere, formation of microbial cenosis etc.) providing an adequate human environment. It is also important that functional reliability of the algal regenerative subsystem is secured by a huge number of cells able, in the event of death of a part of population, to recover in the shortest possible time the size of population and, hence, functionality of the LSS autotrophic component. For a long period of time a Martian crew will be detached from Earth's biosphere and for this reason LSS of their vehicle must be highly reliable, robust and redundant. One of the approaches to LSS redundancy is installation of two systems with different but equally efficient regeneration technologies, i.e. physical-chemical and biological. At best, these two systems should operate in parallel sharing the function of regeneration of the human environment. In case of failure or a sharp deterioration in performance of one system the other will, by way of redundancy, increase its throughput to make up for the loss. This LSS design will enable simultaneous handling of a number of critical problems including adequate satisfaction of human environmental needs.     

The conceptual design of a hybrid life support system based on the evaluation and comparison of terrestrial testbeds.

This report summarizes a trade study of different options of a bioregenerative Life Support System (LSS) and a subsequent conceptual design of a hybrid LSS. The evaluation was based mainly on the terrestrial testbed projects MELISSA (ESA) and BIOS (Russia). In addition, some methods suggested by the Advanced Life Support Project (NASA) were considered. Computer models, including mass flows were established for each of the systems with the goal of closing system loops to the extent possible. In order to cope with the differences in the supported crew size and provided nutrition, all systems were scaled for supporting a crew of six for a 780 day Mars mission (180 days transport to Mars; 600 days surface period) as given in the NASA Design Reference Mission Scenario [Hoffman, S.J., Kaplan, D.L. Human exploration of Mars: the Reference Mission of the NASA Mars Exploratory Study, 1997]. All models were scaled to provide the same daily allowances, as of calories, to the crew. Equivalent System Mass (ESM) analysis was used to compare the investigated system models against each other. Following the comparison of the terrestrial systems, the system specific subsystem options for Food Supply, Solid Waste Processing, Water Management and Atmosphere Revitalization were evaluated in a separate trade study. The best subsystem technologies from the trade study were integrated into an overall design solution based on mass flow relationships. The optimized LSS is mainly a bioregenerative system, complemented by a few physico-chemical elements, with a total ESM of 18,088 kg, which is about 4 times higher than that of a pure physico-chemical LSS, as designed in an earlier study.     

Management and control of microbial populations' development in LSS of missions of different durations.

The problem of interaction between man and microorganisms in closed habitats is an inextricable part of the whole problem of co-existence between macro- and microorganisms. Concerning the support of human life in closed habitat, we can, conventionally, divide microorganisms, acting in life support system (LSS) into three groups: useful, neutral and harmful. The tasks, for human beings for optimal coexistence with microhabitants seem to be trivial: (1) to increase the activity of useful forms, (2) decrease the activity harmful forms, (3) not allow the neutral forms to become the harmful ones and even to help them to gain useful activity. The task of efficient management and control of microbial population's development in LSS highly depends on mission duration. As for short-term missions without recycling, the proper hygienic procedures are developed. For longer missions, the probability of transformation of the neutral forms into the harmful ones is becoming more dangerous. The LSS for long-term missions are to use cycling-recycling systems, including system with biological recycling. In these systems, microbial populations as regenerative link should be useful and active agents. Some problems of microbial populations control and management are discussed in the paper.     

Functional, regulatory and indicator features of microorganisms in man-made ecosystems.

Functional, regulatory and indicator features of microorganisms in development and functioning of the systems and sustaining stability of three man-made ecosystem types has been studied. 1) The functional (metabolic) feature was studied in aquatic ecosystems of biological treatment of sewage waters for the reducer component. 2) The regulatory feature of bacteria for plants (producer component) was studied in simple terrestrial systems "wheat plants-rhizospheric microorganisms-artificial soil" where the behavior of the system varied with activity of the microbial component. For example with atmospheric carbon dioxide content elevated microbes promote intensification of photosynthesis processes, without binding the carbon in the plant biomass. 3) The indicator feature for the humans (consumer component) was studied in Life Support Systems (LSS). High sensitivity of human microflora to system conditions allowed its use as an indicator of the state of both system components and the entire ecosystem. Grant numbers: N99-04-96017, N15.     

Closed Environment Module – Modularization and extension of the Virtual Habitat

The Virtual Habitat (V-HAB), is a Life Support System (LSS) simulation, created to perform dynamic simulation of LSS’s for future human spaceflight missions. It allows the testing of LSS robustness by means of computer simulations, e.g. of worst case scenarios.     

Food production and nutrition in Biosphere 2: results from the first mission September 1991 to September 1993.

The initial test of the Biosphere 2 agricultural system was to provide a nutritionally adequate diet for eight crew members during a two year closure experiment, 1991-1993. The overall results of that trial are presented in this paper. The 2000 m2 cropping area provided about 80 percent of overall nutritional needs during the two years. Adaptation of the crew to the diet which averaged 2200 calories, 73 g. of protein and 32 g. of fat per person over the course of the two years. The diet was primarily vegetarian, with only small amounts of milk, meat and eggs from the system's domestic animals. The crew experienced 10-20 percent weight loss, most of which occurred in the first six months of the closure reflecting adaptation to the diet and lower caloric intake during that period. Since Biosphere 2 is a tightly sealed system, non-toxic methods of pest and disease control were employed and inedible plant material, domestic animal wastes and human waste-water were processed and nutrients returned to the soil. Crop pests and diseases, especially broad mites and rootknot nematode, reduced yields, and forced the use of alternative crops. Outstanding crops included rice, sweet potato, beets, banana, and papaya. The African pygmy goats were the most productive of the domestic animals. Overall, the agriculture and food processing required some 45% of the crew time.