Comparison of aerobically-treated and untreated crop residue as a source of recycled nutrients in a recirculating hydroponic system.

This study compared the growth of potato plants on nutrients recycled from inedible potato biomass. Plants were grown for 105 days in recirculating, thin-film hydroponic systems containing four separate nutrient solution treatments: (1) modified half-strength Hoagland's (control), 2) liquid effluent from a bioreactor containing inedible potato biomass, 3) filtered (0.2 micrometer) effluent, and 4) the water soluble fraction of inedible potato biomass (leachate). Approximately 50% of the total nutrient requirement in treatments 2-4 were provided (recycled) from the potato biomass. Leachate had an inhibitory effect on leaf conductance, photosynthetic rate, and growth (50% reduction in plant height and 60% reduction in tuber yield). Plants grown on bioreactor effluent (filtered or unfiltered) were similar to the control plants. These results indicated that rapidly degraded, water soluble organic material contained in the inedible biomass, i.e., material in leachate, brought about phytotoxicity in the hydroponic culture of potato. Recalcitrant, water soluble organic material accumulated in all nutrient recycling treatments (650% increase after 105 days), but no increase in rhizosphere microbial numbers was observed.     

Enzyme conversion of lignocellulosic plant materials for resource recovery in a Controlled Ecological Life Support System.

A large amount of inedible plant material composed primarily of the carbohydrate materials cellulose, hemicellulose, and lignin is generated as a result of plant growth in a Controlled Ecological Life-Support System (CELSS). Cellulose is a linear homopolymer of glucose, which when properly processed will yield glucose, a valuable sugar because it can be added directly to human diets. Hemicellulose is a heteropolymer of hexoses and pentoses that can be treated to give a sugar mixture that is potentially a valuable fermentable carbon source. Such fermentations yield desirable supplements to the edible products from hydroponically-grown plants such as rapeseed, soybean, cowpea, or rice. Lignin is a three-dimensionally branched aromatic polymer, composed of phenyl propane units, which is susceptible to bioconversion through the growth of the white rot fungus, Pluerotus ostreatus. Processing conditions, that include both a hot water pretreatment and fungal growth and that lead to the facile conversion of plant polysaccharides to glucose, are presented.     

Recycling of inorganic nutrients for hydroponic crop production following incineration of inedible biomass.

The goal of resource recovery in a regenerative life support system is maintenance of product quality to sure support of reliable and predictable levels of life support function performance by the crop plant component. Further, these systems must be maintained over extended periods of time, requiring maintenance of nutrient solutions to avoid toxicity and deficiencies. The focus of this study was to determine the suitability of the ash product following incineration of inedible biomass as a source of inorganic nutrients for hydroponic crop production. Inedible wheat biomass was incinerated and ash quality characterized. The incinerator ash was dissolved in adequate nitric acid to establish a consistent nitrogen concentration is all nutrient solution treatments. Four experimental nutrient treatments were included: control, ash only, ash supplemented to match the control treatment, and ash only quality formulated with reagent grade chemicals. When nutrient solutions were formulated using only ash following incineration of inedible biomass, a balance in solution is established representing elemental retention following incineration and nutrient proportions present in the original biomass. The resulting solution is not identical to the control. This imbalance resulted in a suppression of crop growth. When the ash is supplemented with reagent grade chemicals to establish the same balance as in the control--growth is identical to the control. The ash appears to carry no phytotoxic materials. Growth in solution formulated with reagent grade chemicals but matching the quality of the ash only treatment resulted in similar growth to that of the ash only treatment. The ash product resulting from incineration of inedible biomass appears to be a suitable form for recycle of inorganic nutrients to crop production.     

Spirulina acceptability trials in rats. A study for the "MELISSA" life-support system.

Groups of five rats were fed for sixteen weeks a slightly deficient diet, supplemented with 0-40% of a dried preparation of the blue-green alga Spirulina as a protein source. Control groups were fed a normal rat diet. No significant differences between groups were found in food intake, growth rate or carbon dioxide production. All animals remained apparently healthy, and had similar organ weights. The study suggests that Spirulina may be used as a protein source in rat diets.     

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.     

Production characteristics of the “higher plants–soil-like substrate” system as an element of the bioregenerative life support system

The study addresses the possibility of long-duration operation of a higher plant conveyor, using a soil-like substrate (SLS) as the root zone. Chufa (Cyperus esculentus L.), radish (Raphanus sativus L.), and lettuce (Lactuca sativa L.) were used as study material. A chufa community consisting of 4 age groups and radish and lettuce communities consisting of 2 age groups were irrigated with a nutrient solution, which contained mineral elements extracted from the SLS. After each harvest, inedible biomass of the harvested plants and inedible biomasses of wheat and saltwort were added to the SLS. The amounts of the inedible biomasses of wheat and saltwort to be added to the SLS were determined based on the nitrogen content of the edible mass of harvested plants. CO₂ concentration in the growth chamber was maintained within the range of 1100–1700 ppm. The results of the study show that higher plants can be grown quite successfully using the proposed process of plant waste utilization in the SLS. The addition of chufa inedible biomass to the SLS resulted in species-specific inhibition of growth of both cultivated crops and microorganisms in the “higher plants – SLS” system. There were certain differences between the amounts of some mineral elements removed from the SLS with the harvested edible biomass and those added to it with the inedible biomasses of wheat and saltwort.     

The effects of composting on the nutritional composition of fibrous bio-regenerative life support systems (BLSS) plant waste residues and its impact on the growth of Nile tilapia (Oreochromis niloticus)

Utilization of bio-regenerative life support systems (BLSS) plant waste residues as a nutritional source by Nile tilapia (Oreochromis niloticus) has proven problematic as a result of high concentrations of fibrous compounds in the plant waste residues. Nutritional improvement of plant waste residues by composting with the oyster mushroom (Pleurotus ostreatus), and the effects on growth and nutrient utilization of Nile tilapia fed such residues were evaluated. Five Nile tilapia (mean weight = 70.9 ± 3.1 g) were stocked in triplicate aquaria and fed one of two experimental diets, cowpea (CP) and composted cowpea (CCP), twice daily for a period of 8 weeks. Composting of cowpea residue resulted in reduced concentrations of nitrogen-free extract, hemi-cellulose and trypsin inhibitor activity, though trypsin inhibitor activity remained high. Composting did not reduce crude fiber, lignin, or cellulose concentrations in the diet. No significant differences (P < 0.05) were observed in weight gain, specific growth rate, survival rate, daily consumption, and food conversion ratio between tilapia fed CP and CCP. These results suggest that P. ostreatus is not a suitable candidate for culture in conjunction with the culture of Nile tilapia. Additional work is needed to determine what, if any, benefit can be obtained from incorporating composted residue as feed for Nile tilapia.     

Hydroponic potato production on nutrients derived from anaerobically-processed potato plant residues.

Bioregenerative methods are being developed for recycling plant minerals from harvested inedible biomass as part of NASA's Advanced Life Support (ALS) research. Anaerobic processing produces secondary metabolites, a food source for yeast production, while providing a source of water soluble nutrients for plant growth. Since NH4-N is the nitrogen product, processing the effluent through a nitrification reactor was used to convert this to NO3-N, a more acceptable form for plants. Potato (Solanum tuberosum L.) cv. Norland plants were used to test the effects of anaerobically-produced effluent after processing through a yeast reactor or nitrification reactor. These treatments were compared to a mixed-N treatment (75:25, NO3:NH4) or a NO3-N control, both containing only reagent-grade salts. Plant growth and tuber yields were greatest in the NO3-N control and yeast reactor effluent treatments, which is noteworthy, considering the yeast reactor treatment had high organic loading in the nutrient solution and concomitant microbial activity.     

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.     

Integration of waste processing and biomass production systems as part of the KSC Breadboard project.

After initial emphasis on large-scale baseline crop tests, the Kennedy Space Center (KSC) Breadboard project has begun to evaluate long-term operation of the biomass production system with increasing material closure. Our goal is to define the minimum biological processing necessary to make waste streams compatible with plant growth in hydroponic systems, thereby recycling nutrients into plant biomass and recovering water via atmospheric condensate. Initial small and intermediate-scale studies focused on the recycling of nutrients contained in inedible plant biomass. Studies conducted between 1989-1992 indicated that the majority of nutrients could be rapidly solubilized in water, but the direct use of this crop "leachate" was deleterious to plant growth due to the presence of soluble organic compounds. Subsequent studies at both the intermediate scale and in the large-scale Biomass Production Chamber (BPC) have indicated that aerobic microbiological processing of crop residue prior to incorporation into recirculating hydroponic solutions eliminated any phytotoxic effect, even when the majority of the plant nutrient demand was provided from recycled biomass during long term studies (i.e. up to 418 days). Current and future studies are focused on optimizing biological processing of both plant and human waste streams.     

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.     

Biological degradation and composition of inedible sweetpotato biomass.

Many challenges are presented by biological degradation in a bioregenerative Controlled Ecological Life Support System (CELSS) as envisioned by the U.S. National Aeronautics and Space Administration (NASA). In the studies conducted with biodegradative microorganism indigenous to sweetpotato fields, it was determined that a particle size of 75 microns and incubation temperature of 30 degrees C were optimal for degradation. The composition of the inedible biomass and characterization of plant nutrient solution indicated the presence of potential energy sources to drive microbial transformations of plant waste. Selected indigenous soil isolates with ligno-cellulolytic or sulfate-reducing ability were utilized in biological studies and demonstrated diversity in ability to reduce sulfate in solution and to utilize alternative carbon sources: a lignin analog--4-hydroxy, 3-methoxy cinnamic acid, cellulose, arabinose, glucose, sucrose, mannitol, galactose, ascorbic acid.     

Excess nutrients in hydroponic solutions alter nutrient content of rice, wheat, and potato.

Environment has significant effects on the nutrient content of field-grown crop plants. Little is known, however, about compositional changes caused by controlled environments in which plants receive only artificial radiation and soilless, hydroponic culture. This knowledge is essential for developing a safe, nutritious diet in a Controlled Ecological Life-Support System (CELSS). Three crops that are candidates for inclusion in a CELSS (rice, wheat, and white potato) were grown both in the field and in controlled environments where the hydroponic nutrient solution, photosynthetic photon flux (PPF), and CO2 level were manipulated to achieve rapid growth rates. Plants were harvested at maturity, separated into discrete parts, and dried prior to analysis. Plant materials were analyzed for proximate composition (protein, fat, ash, and carbohydrate), total nitrogen (N), nitrate, minerals, and amino-acid composition. The effect of environment on nutrient content varied by crop and plant part. Total N and nonprotein N (NPN) contents of plant biomass generally increased under controlled-environment conditions compared to field conditions, especially for leafy plant parts and roots. Nitrate levels were increased in hydroponically-grown vegetative tissues, but nitrate was excluded from grains and tubers. Mineral content changes in plant tissue included increased phosphorus and decreased levels of certain micronutrient elements under controlled-environment conditions. These findings suggest that cultivar selection, genetic manipulation, and environmental control could be important to obtain highly nutritious biomass in a CELSS.     

Controlled environments alter nutrient content of soybeans.

Information about compositional changes in plants grown in controlled environments is essential for developing a safe, nutritious diet for a Controlled Ecomological Life-Support System (CELSS). Information now is available for some CELSS candidate crops, but detailed information has been lacking for soybeans. To determine the effect of environment on macronutrient and mineral composition of soybeans, plants were grown both in the field and in a controlled environment where the hydroponic nutrient solution, photosynthetic flux (PPF), and CO2 level were manipulated to achieve rapid growth rates. Plants were harvested at seed maturity, separated into discrete parts, and oven dried prior to chemical analysis. Plant material was analyzed for proximate composition (moisture, protein, lipid, ash, and carbohydrate), total nitrogen (N), nonprotein N (NPN), nitrate, minerals, amino acid composition, and total dietary fiber. The effect of environment on composition varied by cultivar and plant part. Chamber-grown plants generally exhibited the following characteristics compared with field-grown plants: 1) increased total N and protein N for all plant parts, 2) increased nitrate in leaves and stems but not in seeds, 3) increased lipids in seeds, and 4) decreased Ca:P ratio for stems, pods, and leaves. These trends are consistent with data for other CELSS crops. Total N, protein N, and amino acid contents for 350 ppm CO2 and 1000 ppm CO2 were similar for seeds, but protein N and amino acid contents for leaves were higher at 350 ppm CO2 than at 1000 ppm CO2. Total dietary fiber content of soybean leaves was higher with 350 ppm CO2 than with 1000 ppm CO2. Such data will help in selecting of crop species, cultivars, and growing conditions to ensure safe, nutritious diets for CELSS.     

Element exchange in a water- and gas-closed biological Life Support System.

Liquid human wastes and household water used for nutrition of wheat made possible to realize 24% closure for the mineral exchange in an experiment with a 2-component version of "Bios-3" life support system (LSS) Input-output balances of revealed, that elements (primarily trace elements) within the system. The structural materials (steel, titanium), expanded clay aggregate, and catalytic furnace catalysts. By the end of experiment, the permanent nutrient solution, plants, and the human diet gradually built up Ni, Cr, Al, Fe, V, Zn, Cu, and Mo. Thorough selection and pretreatment of materials can substantially reduce this accumulation. To enhance closure of the mineral exchange involves processing of human-metabolic wastes and inedible biomes inside LSS. An efficient method to oxidize wastes by hydrogen peroxide icon a quartz reactor at the temperature of 80 degrees C controlled electromagnetic field is proposed.     

Analysis of nitrogen flow in rice cultivation in CEEF.

In order to control the material circulation in the Closed Ecology Experiment Facilities (CEEF), it is necessary to clarify material flow in the Closed Plant Experiment Facility (CPEF) of CEEF. We tried to grow rice plants and measure the nitrogen contents in rice plant and nutrient solution in plant cultivation bed to trace the material balance in CPEF. The measurements were carried out under the condition of 750 ppm (v/v) CO2 at 26/19 degrees C in the plant cultivation room. The measurements showed the absorbed nitrogen amount in plant was less than the outflow nitrogen amount from nutrient solution. This difference between absorbed and outflow quantity reached to 17%.     

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.     

Low light intensity effects on the growth,photosynthetic characteristics,antioxidant capacity,yield and quality of wheat (Triticum aestivum L.) at different growth stages in BLSS

Minimizing energy consumption and maximizing crop productivity are major challenges to growing plants in Bioregenerative Life Support System (BLSS) for future long-term space mission. As a primary source of energy, light is one of the most important environmental factors for plant growth. The purpose of this study is to investigate the effects of low light intensity at different stages on growth, pigment composition, photosynthetic efficiency, biological production and antioxidant defence systems of wheat (Triticum aestivum L.) cultivars during ontogenesis. Experiments were divided into 3 intensity-controlled stages according to growth period (a total of 65 days): seedling stage (first 20 days), heading and flowering stage (middle 30 days) and grain filling stage (last 15 days). Initial light condition of the control was 420 μmol m⁻² s⁻¹ and the light intensity increased with the growth of wheat plants. The light intensities of group I and II at the first stage and the last stage were adjusted to the half level of the control respectively. For group III, the first and the last stage were both adjusted to half level of the control. During the middle 30 days, all treatments were kept the same intensity. The results indicated that low-light treatment at seedling stage, biomass, nutritional contents, components of inedible biomass and healthy index (including peroxidase (POD) activity, malondialdehyde (MDA) and proline content) of wheat plants have no significant difference to the control. Furthermore, unit kilojoule yield of group I reached 0.591 × 10⁻³ g/kJ and induced the highest energy efficiency. However, low-light treatment at grain filling stage affected the final production significantly.     

Use of biologically reclaimed minerals for continuous hydroponic potato production in a CELSS.

Plant-derived nutrients were successfully recycled in a Controlled Ecological Life Support System (CELSS) using biological methods. The majority of the essential nutrients were recovered by microbiologically treating the plant biomass in an aerobic bioreactor. Liquid effluent containing the nutrients was then returned to the biomass production component via a recirculating hydroponic system. Potato (Solanum tuberosum L.) cv. Norland plants were grown on those nutrients in either a batch production mode (same age plants on a nutrient solution) or a staggered production mode (4 different ages of plants on a nutrient solution). The study continued over a period of 418 days, within NASA Breadboard Project's Biomass Production Chamber at the Kennedy Space Center. During this period, four consecutive batch cycles (104-day harvests) and 13 consecutive staggered cycles (26-day harvests) were completed using reclaimed minerals and compared to plants grown with standard nutrient solutions. All nutrient solutions were continually recirculated during the entire 418 day study. In general, tuber yields with reclaimed minerals were within 10% of control solutions. Contaminants, such as sodium and recalcitrant organics tended to increase over time in solutions containing reclaimed minerals, however tuber composition was comparable to tubers grown in the control solutions.     

ASTROCULTURE (TM) root metabolism and cytochemical analysis.

Physiology of the root system is dependent upon oxygen availability and tissue respiration. During hypoxia nutrient and water acquisition may be inhibited, thus affecting the overall biochemical and physiological status of the plant. For the Astroculture (TM) plant growth hardware, the availability of oxygen in the root zone was measured by examining the changes in alcohol dehydrogenase (ADH) activity within the root tissue. ADH activity is a sensitive biochemical indicator of hypoxic conditions in plants and was measured in both spaceflight and control roots. In addition to the biochemical enzyme assays, localization of ADH in the root tissue was examined cytochemically. The results of these analyses showed that ADH activity increased significantly as a result of spaceflight exposure. Enzyme activity increased 248% to 304% in dwarf wheat when compared with the ground controls and Brassica showed increases between 334% and 579% when compared with day zero controls. Cytochemical staining revealed no differences in ADH tissue localization in any of the dwarf wheat treatments. These results show the importance of considering root system oxygenation in designing and building nutrient delivery hardware for spaceflight plant cultivation and confirm previous reports of an ADH response associated with spaceflight exposure.