Integrating biological treatment of crop residue into a hydroponic sweetpotato culture.

Residual biomass from hydroponic culture of sweetpotato [Ipomoea batatas (L.) Lam.] was degraded using natural bacterial soil isolates. Sweetpotato was grown for 120 days in hydroponic culture with a nutrient solution comprised of a ratio of 80% modified half Hoagland solution to 20% filtered effluent from an aerobic starch hydrolysis bioreactor. The phytotoxicity of the effluent was assayed with Waldmann's Green' lettuce (Lactuca sativa L.) and the ratio selected after a 60-day bioassay using sweetpotato plants propagated vegetatively from cuttings. Controlled environment chamber experiments were conducted to investigate the impact of filtrate from biological treatment of crop residue on growth and storage root production with plants grown in a modified half Hoagland solution. Incorporation of bioreactor effluent, reduced storage root yield of 'Georgia Jet' sweetpotato but the decrease was not statistically significant when compared with yield for plants cultured in a modified half Hoagland solution without filtrate. However, yield of 'TU-82-155' sweetpotato was significantly reduced when grown in a modified half Hoagland solution into which filtered effluent had been incorporated. Total biomass was significantly reduced for both sweetpotato cultivars when grown in bioreactor effluent. The leaf area and dry matter accumulation were significantly (P < 0.05) reduced for both cultivars when grown in solution culture containing 20% filtered effluent.     

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.     

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.     

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.     

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.     

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.     

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.     

Theoretical and practical considerations for staggered production of crops in a BLSS.

A functional Bioregenerative Life Support System (BLSS) will generate oxygen, remove excess carbon dioxide, purify water, and produce food on a continuous basis for long periods of operation. In order to minimize fluctuations in gas exchange, water purification, and yield that are inherent in batch systems, staggered planting and harvesting of the crop is desirable. A 418-d test of staggered production of potato cv. Norland (26-d harvest cycles) using nutrients recovered from inedible biomass was recently completed at Kennedy Space Center. The results indicate that staggered production can be sustained without detrimental effects on life support functions in a CELSS. System yields of H2O, O2 and food were higher in staggered than batch plantings. Plants growing in staggered production or batch production on "aged" solution initiated tubers earlier, and were shorter than plants grown on "fresh" solution. This morphological response required an increase in planting density to maintain full canopy coverage. Plants grown in staggered production used available light more efficiently than the batch planting due to increased sidelighting.     

Anaerobic degradation of inedible crop residues produced in a Controlled Ecological Life Support System.

An anaerobic reactor seeded with organisms from an anaerobic lagoon was used to study the degradation of inedible crop residues from potato and wheat crops grown in a closed environment. Conversion of this biomass into other products was also evaluated. Degradation of wheat volatile solids was about 25% where that of potato was about 50%. The main product of the anaerobic fermentation of both crops was acetic acid with smaller quantities of propionate and butyrate produced. Nitrate, known to be high in concentration in inedible potato and wheat biomass grown hydroponically, was converted to ammonia in the anaerobic reactor. Both volatile fatty acid and ammonia production may have implications in a crop production system.     

Proximate composition of CELSS crops grown in NASA's Biomass Production Chamber.

Edible biomass from four crops of wheat (Triticum aestivum L.), four crops of lettuce (Lactuca sativa L.), four crops of potato (Solanum tuberosum L.), and three crops of soybean (Glycine max (L.) Merr.) grown in NASA's CELSS Biomass Production Chamber were analyzed for proximate composition. All plants were grown using recirculating nutrient (hydroponic) film culture with pH and electrical conductivity automatically controlled. Temperature and humidity were controlled to near optimal levels for each species and atmospheric carbon dioxide partial pressures were maintained near 100 Pa during the light cycles. Soybean seed contained the highest percentage of protein and fat, potato tubers and wheat seed contained the highest levels of carbohydrate, and lettuce leaves contained the highest level of ash. Analyses showed values close to data published for field-grown plants with several exceptions: In comparison with field-grown plants, wheat seed had higher protein levels; soybean seed had higher ash and crude fiber levels; and potato tubers and lettuce leaves had higher protein and ash levels. The higher ash and protein levels may have been a result of the continuous supply of nutrients (e.g., potassium and nitrogen) to the plants by the recirculating hydroponic culture.     

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.     

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.     

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.     

The nutritional adequacy of a limited vegan diet for a Controlled Ecological Life-Support System.

Purdue University, as well as the Johnson and Kennedy Space Centers and NASA Ames Research Center, are investigating approximately 5-10 plants that will be grown hydroponically to provide not only the energy and nutrients, but also the oxygen for humans habitating in Mars and lunar bases. The growth and nutritional status of rats fed either a control diet (adequate in all macro- and micronutrients) or a strict vegetarian diet consisting of 5 (vegan-5) or 10 (vegan-10) candidate crop species were investigated. In addition, vegan-10 diets were supplemented with mineral and/or vitamin mix at a level similar to the control diets to assess the effect of supplementation on nutrient status. The assessment of inedible plant material as an alternative food source was also investigated. Results of this study demonstrated that consumption of the vegan-10 diet significantly improved weight gain of rats compared to that for rats fed the vegan-5 diet. Mineral supplementation, at a level present in the control diet, to the vegan-10 diet improved growth and nutrient status, but growth was significantly lower compared to the control-fed rats. Inclusion of inedible plant material, high in ash content, improved some indices of nutrient status, without improving growth.     

Potato growth in a porous tube water and nutrient delivery system.

Potato (Solanum tuberosum L.) cv. 'Norland', vegetative growth and tuber productivity grown in the porous water and nutrient delivery system (PTNDS) developed by the Wisconsin Center for Space Automation and Robotics were compared with the vegetative growth and tuber productivity of plants grown in a peat:vermiculite potting mixture (PT/VR). The plants were grown at 12, 16, and 24-h light periods, 18 degrees C constant temperature, 70% relative humidity, and 300 micromol m-2 s-1 photosynthetic photon flux. Canopy height of plants grown in the PT/VR system was taller than that of plants grown in the PTNDS system. Canopy height differences were greatest when the plants were grown under a 24-h photoperiod. Leaf and stem dry masses were similar for plants grown in the two systems under the 12-h photoperiod. Under the 24-h photoperiod, leaf and stem dry masses of plants grown in the PT/VR system were more than 3 times those of plants grown in the PTNDS system. Tuber dry masses were similar for plants grown in the two systems under the 12-h photoperiod. Under the 24 h-photoperiod, tuber dry weights of plants grown in the PT/VR system were more than twice those of plants grown in the PTNDS system. A slightly higher harvest index (ratio of tuber weight to leaf plus stem weight) was noted for the plants grown in the PTNDS than for the plants grown in the PT/VR system. Plants grown in the PTNDS system at the 24-h photoperiod matured earlier than plants grown at this photoperiod in the PT/VR system. Vegetative growth and tuber productivity of plants grown under the 16-h photoperiod generally were intermediate to those noted for plants grown under the 12 and 24-h photoperiods. These results indicate that potato plants grown in a PTNDS system may require less plant growing volume, mature in a shorter time, and likely produce more tubers per unit area compared with plants grown in the PT/VR system. These plant characteristics are a distinct advantage for a plant growing unit of a CELSS.     

NASA's Biomass Production Chamber: a testbed for bioregenerative life support studies.

The Biomass Production Chamber (BPC) located at Kennedy Space Center, FL, USA provides a large (20 m2 area, 113 m3 vol.), closed environment for crop growth tests for NASA's Controlled Ecological Life Support System (CELSS) program. Since the summer of 1988, the chamber has operated on a near-continuous basis (over 1200 days) without any major failures (excluding temporary power losses). During this time, five crops of wheat (64-86 days each), three crops of soybean (90 to 97 days), five crops of lettuce (28-30 days), and four crops of potato (90 to 105 days were grown, producing 481 kg of dry plant biomass, 196 kg edible biomass, 540 kg of oxygen, 94,700 kg of condensed water, and fixing 739 kg of carbon dioxide. Results indicate that total biomass yields were close to expected values for the given light input, but edible biomass yields and harvest indices were slightly lower than expected. Stand photosynthesis, respiration, transpiration, and nutrient uptake rates were monitored throughout growth and development of the different crops, along with the build-up of ethylene and other volatile organic compounds in the atmosphere. Data were also gathered on system hardware maintenance and repair, as well as person-hours required for chamber operation. Future tests will include long-term crop production studies, tests in which nutrients from waste treatment systems will be used to grow new crops, and multi-species tests.     

Utilization of sweet potatoes in controlled ecological life support systems (CELSS).

A number of studies have selected the sweet potato as a potentially important crop for CELSS. Most hydroponic studies of sweet potatoes have been short term (<80 days). Full term (90 to 150 days) studies of sweet potatoes in hydroponic systems were needed to understand the physiology of storage root enlargement and to evaluate sweet potato production potential for CELSS. Early and late maturing sweet potato varieties were crown in hydroponic systems of different types--static with periodic replacement, flowing with and without recirculation, aggregate, and non-aggregate. In a flowing system with recirculation designed at Tuskegee University using the nutrient film technique (NFT), storage root yields as high as 1790 g were produced with an edible growth rate of up to 66 g m-2 d-1 and a harvest index as high as 89% under greenhouse conditions. Preliminary experiments indicated high yields can be obtained in controlled environmental chambers. Significant cultivar differences were found in all systems studied. Nutritive composition of storage roots and foliage were similar to field-grown plants. The results indicate great potential for sweet potato in CELSS.     

Selection and hydroponic growth of potato cultivars for bioregenerative life support systems

As part of the ESA-funded MELiSSA program, Ghent University and the Université catholique de Louvain investigated the suitability, growth and development of four potato cultivars in hydroponic culture under controlled conditions with the aim to incorporate such cultivation system in an Environmental Control and Life Support System (ECLSS). Potato plants can fulfill three major functions in an ECLSS in space missions: (a) fixation of CO₂ and production of O₂, (b) production of tubers for human nutrition and (c) production of clean water after condensation of the water vapor released from the plants by transpiration. Four cultivars (Annabelle, Bintje, Desiree and Innovator) were selected and grown hydroponically in nutrient film technique (NFT) gullies in a growth chamber under controlled conditions. The plant growth parameters, tuber harvest parameters and results of tuber nutritional analysis of the four cultivars were compared. The four potato cultivars grew well and all produced tubers. The growth period lasted 127 days for all cultivars except for Desiree which needed 145 days. Annabelle (1.45 kg/m²) and Bintje (1.355 kg/m²) were the best performing of the four cultivars. They also produced two times more tubers than Desiree and Innovator. Innovator produced the biggest tubers (20.95 g/tuber) and Desiree the smallest (7.67 g/tuber). The size of Annabelle and Bintje potatoes were intermediate. Bintje plants produced the highest total biomass in term of DW. The highest non-edible biomass was produced by Desiree, which showed both the highest shoot and root DW. The manual length and width measurements were also used to predict the total tuber mass. The energy values of the tubers remained in the range of the 2010 USDA and Souci-Fachmann-Kraut food composition databases. The amount of Ca determined was slightly reduced compared to the USDA value, but close to the Souci-Fachmann-Kraut value. The concentration of Cu, Zn and P were high compared to both databases.     

Incineration of biomass and utilization of product gas as a CO2 source for crop production in closed systems: gas quality and phytotoxicity.

This study addressed the recycle of carbon from inedible biomass to CO2 for utilization in crop production. Earlier work identified incineration as an attractive approach to resource recovery from solid wastes because the products are well segregated. Given the effective separation of carbon into the gaseous product stream from the incinerator in the form of CO2 we captured the gaseous stream produced during incineration of wheat inedible biomass and utilized it as the CO2 source for crop production. Injection rate was based on maintenance of CO2 concentration in the growing environment. The crop grown in the closed system was lettuce. Carbon was primarily in the form of CO2 in the incinerator product gas with less than 8% of carbon compounds appearing as CO. Nitrogen oxides and organic compounds such as toluene, xylene, and benzene were present in the product gas at lower concentrations (< 4 micromol mol-1); sulfur containing compounds were below the detection limits. Direct utilization of the gaseous product of the incinerator as the CO2 source was toxic to lettuce grown in a closed chamber. Net photosynthetic rates of the crop was suppressed more than 50% and visual injury symptoms were visible within 3 days of the introduction of the incinerator gas. Even the removal of the incinerator gas alter two days of crop exposure and replacement with pure CO2 did not eliminate the toxic effects. Both organic and inorganic components of the incinerator gas are candidates for the toxin.     

Significance of rhizosphere microorganisms in reclaiming water in a CELSS.

Plant-microbe interactions, such as those of the rhizosphere, may be ideally suited for recycling water in a Controlled Ecological Life Support System (CELSS). The primary contaminant of waste hygiene water will be surfactants or soaps. We identified changes in the microbial ecology in the rhizosphere of hydroponical1y grown lettuce during exposure to surfactant. Six week old lettuce plants were transferred into a chamber with a recirculating hydroponic system. Microbial density and population composition were determined for the nutrient solution prior to introduction of plants and then again with plants prior to surfactant addition. The surfactant Igepon was added to the recirculating nutrient solution to a final concentration of 1.0 g L-1. Bacteria density and species diversity of the solution were monitored over a 72-h period following introduction of Igepon. Nine distinct bacterial types were identified in the rhisosphere; three species accounted for 87% of the normal rhizosphere population. Microbial cell number increased in the presence of Igepon, however species diversity declined. At the point when Igepon was degraded from solution, diversity was reduced to only two species. Igepon was found to be degraded directly by only one species found in the rhizosphere. Since surfactants are degraded from the waste hygiene water within 24 h, the potential for using rhizosphere bacteria as a waste processor in a CELSS is promising.