Selection of microalgae suitable for culturing with digestate from methane fermentation.

The effects ofdigestate on the growth rates of Euglena gracilis, Chlorella vulgaris, and Dunaliella tertiolecta were investigated to select suitable microalgae for culturing with digestate from methane fermentation. Microalgae were cultured in an aqueous solution containing digestate at concentrations of 5%, 10%, 13%, 20%, 40%, 50%, and 100%, and Cramer-Myers (CM) solution as a control, at photosynthetic photon flux densities (PPFDs) of 75-150 micromol m(-2) s(-1) with continuous illumination at 30 degrees C. The number of cells was monitored daily, and specific growth rates (mu) were calculated as cellular multiplication rates. The maximum mu values of these species were greater in appropriate concentrations of digestate than in CM medium. The maximum mu values were 0.047 h(-1) in 10% digestate for E. gracilis, 0.065 h(-1) in 20% digestate for C. vulgaris, and 0.052 h(-1) in 50% digestate for D. tertiolecta at a PPFD of 150 micromol m(-2) s(-1). The mu of D. tertiolecta were 2.5 and 1.1 times higher than those of E. gracilis and C. vulgaris, respectively, in 50% digestate. These results demonstrated that these species could be cultured at high growth rates with diluted methane fermentation sludge and that, among these species, Dunaliella sp. was suitable for culturing at higher concentration of digestate under relatively low-level light conditions.

Effects of temperature, CO2/O2 concentrations and light intensity on cellular multiplication of microalgae, Euglena gracilis.

Microalgae culture is likely to play an important role in aquatic food production modules in bioregenerative systems for producing feeds for fish, converting CO2 to O2 and remedying water quality as well as aquatic higher plants. In the present study, the effects of culture conditions on the cellular multiplication of microalgae, Euglena gracilis, was investigated as a fundamental study to determine the optimum culture conditions for microalgae production in aquatic food production modules including both microalgae culture and fish culture systems. E. gracilis was cultured under conditions with five levels of temperatures (25-33 degrees C), three levels of CO2 concentrations (2-6%), five levels of O2 concentrations (10-30%), and six levels of photosynthetic photon flux (20-200 micromoles m-2 s-1). The number of Euglena cells in a certain volume of solution was monitored with a microscope under each environmental condition. The multiplication rate of the cells was highest at temperatures of 27-31 degrees C, CO2 concentration of 4%, O2 concentration of 20% and photosynthetic photon flux of about 100 micromoles m-2 s-1. The results demonstrate that E. gracilis could efficiently produce biomass and convert CO2 to O2 under relatively low light intensities in aquatic food production modules.

Effects of CO2 concentration and light intensity on photosynthesis of a rootless submerged plant, Ceratophyllum demersum L., used for aquatic food production in bioregenerative life support systems.

In addition to green microalgae, aquatic higher plants are likely to play an important role in aquatic food production modules in bioregenerative systems for producing feed for fish, converting CO2 to O2 and remedying water quality. In the present study, the effects of culture conditions on the net photosynthetic rate of a rootless submerged plant, Ceratophyllum demersum L., was investigated to determine the optimum culture conditions for maximal function of plants in food production modules including both aquatic plant culture and fish culture systems. The net photosynthetic rate in plants was determined by the increase in dissolved O2 concentrations in a closed vessel containing a plantlet and water. The water in the vessel was aerated sufficiently with a gas containing a known concentration of CO2 gas mixed with N2 gas before closing the vessel. The CO2 concentrations in the aerating gas ranged from 0.3 to 10 mmol mol-1. Photosynthetic photon flux density (PPFD) in the vessel ranged from 0 (dark) to 1.0 mmol m-2 s-1, which was controlled with a metal halide lamp. Temperature was kept at 28 degrees C. The net photosynthetic rate increased with increasing PPFD levels and was saturated at 0.2 and 0.5 mmol m-2 s-1 PPFD under CO2 levels of 1.0 and 3.0 mmol mol-1, respectively. The net photosynthetic rate increased with increasing CO2 levels from 0.3 to 3.0 mmol mol-1 showing the maximum value, 75 nmol O2 gDW-1 s-1, at 2-3 mmol mol-1 CO2 and gradually decreased with increasing CO2 levels from 3.0 to 10 mmol mol-1. The results demonstrate that C. demersum could be an efficient CO2 to O2 converter under a 2.0 mmol mol-1 CO2 level and relatively low PPFD levels in aquatic food production modules.

Effects of air velocity on photosynthesis of plant canopies under elevated CO2 levels in a plant culture system.

To obtain basic data for adequate air circulation for promoting plant growth in closed plant production modules in bioregenerative life support systems in space, effects of air velocities ranging from 0.1 to 0.8 m s-1 on photosynthesis in tomato seedlings canopies were investigated under atmospheric CO2 concentrations of 0.4 and 0.8 mmol mol-1. The canopy of tomato seedlings on a plug tray (0.4 x 0.4 m2) was set in a wind-tunnel-type chamber (0.6 x 0.4 x 0.3 m3) installed in a semi-closed-type assimilation chamber (0.9 x 0.5 x 0.4 m3). The net photosynthetic rate in the plant canopy was determined with the differences in CO2 concentrations between the inlet and outlet of the assimilation chamber multiplied by the volumetric air exchange rate of the chamber. Photosynthetic photon flux (PPF) on the plant canopy was kept at 0.25 mmol m-2 s-1, air temperature at 23 degrees C and relative humidity at 55%. The leaf area indices (LAIs) of the plant canopies were 0.6-2.5 and plant heights were 0.05-0.2 m. The net photosynthetic rate of the plant canopy increased with increasing air velocities inside plant canopies and saturated at 0.2 m s-1. The net photosynthetic rate at the air velocity of 0.4 m s-1 was 1.3 times that at 0.1 m s-1 under CO2 concentrations of 0.4 and 0.8 mmol mol-1. The net photosynthetic rate under CO2 concentrations of 0.8 mmol mol-1 was 1.2 times that under 0.4 mmol mol-1 at the air velocity ranging from 0.1 to 0.8 m s-1. The results confirmed the importance of controlling air movement for enhancing the canopy photosynthesis under an elevated CO2 level as well as under a normal CO2 level in the closed plant production modules.

Plant growth and gas balance in a plant and mushroom cultivation system.

In order to obtain basic data for construction of a plant cultivation system incorporating a mushroom cultivation subsystem in the CELSS, plant growth and atmospheric CO2 balance in the system were investigated. The plant growth was promoted by a high level of CO2 which resulted from the respiration of the mushroom mycelium in the system. The atmospheric CO2 concentration inside the system changed significantly due to the slight change in the net photosynthetic rate of plants and/or the respiration rate of the mushroom when the plant cultivation system combined directly with the mushroom cultivation subsystem.

Development of a plant growth unit for growing plants over a long-term life cycle under microgravity conditions.

To study the effect of the space environment on plant growth including the reproductive growth and genetic aberration for a long-term plant life cycle, we have initiated development of a new type of facility for growing plants under microgravity conditions. The facility is constructed with subsystems for controlling environmental elements. In this paper, the concept of the facility design is outlined. Subsystems controlling air temperature, humidity, CO2 concentration, light and air circulation around plants and delivering recycled water and nutrients to roots are the major concerns. Plant experiments for developing the facility and future plant experiments with the completed facility are also overviewed. We intend to install this facility in the Japan Experiment Facility (JEM) boarded on the International Space Station.

Effects of air current speed on gas exchange in plant leaves and plant canopies.

To obtain basic data on adequate air circulation to enhance plant growth in a closed plant culture system in a controlled ecological life support system (CELSS), an investigation was made of the effects of the air current speed ranging from 0.01 to 1.0 m s-1 on photosynthesis and transpiration in sweetpotato leaves and photosynthesis in tomato seedlings canopies. The gas exchange rates in leaves and canopies were determined by using a chamber method with an infrared gas analyzer. The net photosynthetic rate and the transpiration rate increased significantly as the air current speeds increased from 0.01 to 0.2 m s-1. The transpiration rate increased gradually at air current speeds ranging from 0.2 to 1.0 m s-1 while the net photosynthetic rate was almost constant at air current speeds ranging from 0.5 to 1.0 m s-1. The increase in the net photosynthetic and transpiration rates were strongly dependent on decreased boundary-layer resistances against gas diffusion. The net photosynthetic rate of the plant canopy was doubled by an increased air current speed from 0.1 to 1.0 m s-1 above the plant canopy. The results demonstrate the importance of air movement around plants for enhancing the gas exchange in the leaf, especially in plant canopies in the CELSS.

Carbon dioxide and oxygen budgets of a plant cultural system in a CELSS--a case of cultivation of lettuce and turnips.

In order to collect basic data about CO2 and O2 budgets of a plant cultural system in a CELSS, the variation of the CO2 absorption rates of lettuce and turnips were observed during the growing period, under different conditions. The O2 release rates were deduced from the CO2 absorption rates multiplied by 32/44. As a result, when the light intensity, the photoperiod and the atmospheric CO2 concentration increased, the rates also increased. The effects on the turnips were more significant than those on the lettuce. Turnips at 310 micromoles/m2/s of PPFD, 24 hours of photoperiod and 1100 ppm of CO2 concentration grew most actively in the present experimental conditions. One turnip absorbed 32.3 g CO2 and released 23.5 g O2 for 6 days between 24 days and 30 days after sowing.

Ground performance of air conditioning and water recycle system for a Space Plant Box.

Researchers from 5 Japanese universities have developed a plant growth facility (Space Plant Box) for seed to seed experiments under microgravity. The breadboard model of the Space Plant Box was fabricated by assembling subsystems developed for microgravity. The subsystems include air conditioning and water recycle system, air circulation system, water and nutrient delivery system, lighting system and plant monitoring system. The air conditioning and water recycle system is simply composed of a single heat exchanger, two fans and hydrophilic fibrous strings. The strings allow water movement from the cooler fin in the Cooling Box to root supporting materials in the Plant Growth Chamber driven by water potential deficit. Relative humidity in the Plant Growth Chamber can be changed over a wide range by controlling the ratio of latent heat exchange to sensible heat exchange on the cooling fin of the heat exchanger. The transpiration rate was successfully measured by circulating air inside the Plant Growth Chamber only. Most water was recycled and a small amount of water needed to be added from the outside. The simple, air conditioning and water recycle system for the Space Plant Box showed good performance through a barley (Hordeum vulgare L.) growth experiment.

The effect of gravity on surface temperature and net photosynthetic rate of plant leaves.

To clarify the effects of gravity on heat/gas exchange between plant leaves and the ambient air, the leaf temperatures and net photosynthetic rates of plant leaves were evaluated at 0.01, 1.0, 1.5 and 2.0 G of 20 seconds each during a parabolic airplane flight. Thermal images of leaves were captured using infrared thermography at an air temperature of 26 degrees C, a relative humidity of 15% and an irradiance of 260 W m-2. The net photosynthetic rates were determined by using a chamber method with an infrared gas analyzer at an air temperature of 20 degrees C, a relative humidity of 50% and a photosynthetic photon flux of 0.5 mmol m-2 s-1. The mean leaf temperature increased by 1 degree C and the net photosynthetic rate decreased by 13% with decreasing gravity levels from 1.0 to 0.01 G. The leaf temperature decreased by 0.5 degree C and the net photosynthetic rate increased by 7% with increasing gravity levels from 1.0 to 2.0 G. Heat/gas exchanges between leaves and the ambient air were more retarded at lower gravity levels. A restricted free air convection under microgravity conditions in space would limit plant growth by retarding heat and gas exchanges between leaves and the ambient air.

Effects of light intensity and air velocity on air temperature, water vapor pressure, and CO2 concentration inside a plant canopy under an artificial lighting condition.

In order to characterize environmental variables inside a plant canopy under artificial lighting in the CELSS, we investigated the effects of light intensity and air velocity on air temperature, water vapor pressure, and CO2 concentration inside a plant canopy. Under a PPF of 500 micromoles m-2 s-1, air temperature was 2-3 degrees C higher, water vapor pressure was 0.6 kPa higher, and CO2 concentration was 25-35 micromoles mol-1 lower at heights ranging from 0 to 30 mm below the canopy than at a height 60 mm above the canopy. Increasing the PPF increased air temperature and water vapor pressure and decreased CO2 concentration inside the canopy. The air temperature was lower and the CO2 concentration was higher inside the canopy at an air velocity of 0.3 m s-1 than at an air velocity of 0.1 m s-1. The environmental variables inside the canopy under a high light intensity were characterized by higher air temperature, higher vapor pressure, and lower CO2 concentration than those outside the canopy.

Performance of a water suction system using hydrophilic fibrous cloth under low gravity and microgravity in parabolic flight.

For suction of water from a water supply vessel including both water and air under microgravity and g-jitter conditions, a water suction system using hydrophilic fibrous cloth was developed and its performance was evaluated at 0.01-0.02 g-realized for 20 s by parabolic flight in an aircraft. Vessels used for the experiment were glass flasks and had a suction port for suction filtration. A piece of hydrophilic fibrous cloth was arranged along the inner surface of the vessels and the end was fixed to the suction port of the vessels. In vessel without hydrophilic cloths and containing 220 mL of water, the water did not move more than 5 mm along the inner surface and did not reach the suction port under low gravity. When hydrophilic cloths were used, on the other hand, water gathered onto the cloth surface, moved up along the cloth and reached the suction port under low gravity. The amount of water sucked from vessels varied with the amount of water in the vessel and the sectional area of hydrophilic cloths. When the vessels including both water and air were flown during parabolic flight (10(-4) g), water in the vessel moved along the cloth and a water film was formed on the cloth. These results indicated that it is possible to suck water using the fibrous cloth suction system under low gravity and microgravity conditions. Under low gravity conditions, it was difficult to suck water only. However, it is not necessary to separate water from air when the system is used for supplying water to plant root medium consisting of both liquid and gas phases.

Effects of CO2 and O2 concentrations and light intensity on growth of microalgae (Euglena gracilis) in CELSS.

Green microalgae are likely to play an important role in bioregenerative systems for producing food and converting CO2 to O2 in a controlled ecological life support system (CELSS). In the present study, a method for evaluating the effects of environmental variables on the multiplication rate of microalgal cells was developed to determine the optimum culture condition for a microalgal culture system that can function effectively in the CELSS. The microalga, Euglena gracilis, was cultured in water droplets (3 microliters in liquid volume each) in a vessel (25 ml in air volume) in which the CO2 and O2 concentrations were controlled. The number of Euglena cells cultured at CO2 concentrations ranging from 2% to 6%, O2 concentrations ranging from 5% to 20%, and PPF levels ranging from 50 to 100 micromoles m-2 s-1 was monitored by using a video camera and a microscope. The multiplication rate of cells was highest and the cell number increased by 8.3 times during 48 h under a condition of 4% CO2, 21% O2 and 100 micromoles m-2 s-1 PPF. The multiplication rate of the cells was highest at 4% CO2, followed by 6% and 2% CO2, and it decreased with decreasing O2 concentration and decreasing PPF.

The effect of gravity on surface temperatures of plant leaves.

A fundamental study was conducted to develop a facility having an adequate air circulation system for growing healthy plants over a long-term under microgravity conditions in space. To clarify the effects of gravity on heat exchange between plant leaves and the ambient air, surface temperatures of sweet potato and barley leaves and replica leaves made of wet paper and copper were evaluated at gravity levels of 0.01, 1.0, 1.5 and 2.0 g for 20 s each during parabolic aeroplane flights. Thermal images were captured using infrared thermography at an air temperature of 26 degrees C, a relative humidity of 18% and an irradiance of 260 W m-2. Mean leaf temperatures increased by 0.9-1.0 degrees C with decreasing gravity levels from 1.0 to 0.01 g and decreased by 0.5 degrees C with increasing gravity levels from 1.0 to 2.0 g. The increase in leaf temperatures was at most 1.9 degrees C for sweet potato leaves over 20 s as gravity decreased from 1.0 to 0.01 g. The boundary layer conductance to sensible heat exchange decreased by 5% when the gravity decreased from 1.0 to 0.01 g at the air velocity of 0.2 m s-1. The decrease in the boundary layer conductance with decrease in the gravity levels was more significant in a lower air velocity. Heat exchange between leaves and the ambient air was more retarded at lower gravity levels because of less sensible and latent heat transfers with less heat convection.