National Space Development Agency of Japan (NASDA) has been developed aquatic animal experiment facilities for space experiments using NASA Space Shuttle. Vestibular Function Experiment Unit (VFEU) has been firstly designed and developed for Spacelab-J mission (STS-47), and 8 days space experiment with carp has been performed. Following the VFEU, Aquatic Animal Experiment Unit (AAEU) has been developed to accommodate small aquatic animals for second International Microgravity Laboratory mission (IML-2, STS-65). Four kinds of space experiments with goldfish, medaka, newt, and newt eggs have been performed for 15 days mission duration. Then, VFEU has been improved to accommodate marine fish under low temperature condition for Neurolab (STS-90) and STS-95 missions. 17days (STS-90) and 9days (STS-95) experiments with oyster toadfish have been performed by using the VFEU. This report summarizes the outline of these aquatic animal experiment facilities.

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In STS-95 Space Shuttle mission, an aquatic animal research facility, Vestibular Function Experiment Unit (VFEU), was flown to perform neurobiological experiment with marine fish, oyster toadfish. For this purpose, we have developed a sea water purification system using highly active nitrifying bacteria at low temperature. With this system, the water quality in the VFEU was maintained in sufficient condition to keep the toadfish in healthy state for 9 days of the mission. This report summarizes the efficiency of the filter system based on the results from pre-flight bacterial preparation, water analysis of samples taken during flight, and the post-flight analysis of the bacterial filter.

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Vestibular Function Experiment Unit (VFEU), one of the Spacelab facility flown in Neurolab mission (STS-90) in April, 1998, was to support neurophysiological research using a marine fish, Opsanus tau (oyster toadfish). The functions of the VFEU were primarily a quality management of environmental water during the mission at 14°C and for acquiring physiological signals from implanted micro-electrodes in the otolith nerves as well as the spatial acceleration of the fish. A key element of the life support system was a balanced biological filter containing two types of nitrifying bacteria, Nitrosomonas for ammonia oxidization and Nitrobacter for nitrite oxidization. Although the 16 days mission was successful, two toadfishes died in late phase of the mission. Ammonium concentration in those two life support systems elevated to remarkably high level at the end of the mission whereas the other two indicated very low. This report summarizes the results of the water quality management of the VFEU during the Neurolab mission based on analysis of water samples taken during the flight and those taken prior and just after the flight.

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In a space environment such as Space Shuttle or Space Station, animal experiments with aquatic species in a closed system pose a crucial problem in maintaining their water quality for a long term. In nature, ammonia as an animal wastes is converted by nitrifying bacteria to nitrite or nitrate compounds, which usually become nitrogen sources for plants. Thus an application of the biological reactor with such bacteria attached on some filters has been suggested and experimentally studied for efficient waste managements of ammonia. Although some successful results were reported (Kozu et al. 1995, Nagaoka et al. 1998, Nakamura et al. 1997, 1998) in the space applications, purely empirical approaches have so far been taken to develop a biological filter having a stable nitrifying activity. In this study, we constructed a mathematical model to deal with the dynamics of the ammonia nitrifying processes in a biological reactor. The model describes population dynamics of the ammonia-oxidizing bacteria and the nitrite-oxidizing bacteria cultivated on the same filter. We estimated parameters involved in the model using the experimental data. The result shows that these estimated parameters could be applied to general cases and that the two bacteria are in a symbiotic relationship; they can better perform when both coexist, as has been empirically recognized. Based on the model analysis, we discuss how to prepare a high performance biological filter.

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Five crucian carp, Carassius auratus langsdorfiicarps had been reared in a closed water recirculating system. The system was equipped with the compact bioreactor using the plate gels capable of both nitrification and denitrification in a single unit. Ammonia and nitrite concentrations in the rearing water had been maintained below 0.05 mg-N⁄L, and nitrate concentration also controlled between 2 and 8 mg-N⁄L with the bioreactor. As concerns nitrogen budget in the closed system, 95.0 % of nitrogen income from feed was lost as nitrogen gas from the closed system. All fish was alive for 91 days without any unusual behavior. Thus, the bioreactor performed both nitrification and denitrification abilities enough to rear the five fish for 91 days. The bioreactor using the plate gels would be effective to simplify the closed system both physically and operationally, since it can remove the ammonia excreted from fish as nitrogen gas by a single step.

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We have developed the long-term life support system that enables the experiment of aquatic animals breeding for 90 days or more for the future experiments in orbit. In order to enable long-term breeding of wide aquatic animals, it is necessary to remove nitrate produced by biological nitrification. Then, we examined a denitrification method to use an electrochemical reaction of biofilm-electrode reactor. In this research, we have not kept the aquatic animals actually but imitated breeding of five goldfish. The ammonia of about 250 ppm was added in breeding water through 90 days. As a result, neither ammonia nor nitrite accumulated 0.1 ppm and nitrate could be suppressed to about 10 ppm.

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Since a biological filter with nitrifying bacteria was firstly applied to aquatic animal experiments in IML-2 mission, the reactor system has been further studied to combine both nitrifying and denitrifying reactions under aerobic environment allowing an efficient removal of inorganic nitrogen from animal wastes. The isolated denitrifying bacteria had an activity under aerobic condition with rice straw providing a metabolic carbon source for the reaction. The advantage of the aerobic biological filter having both nitrifying and denitrifying activities may allow to reduce the size of the life support system and also for its manageability. The paper reports characteristics of the biological filter systems used for the IML-2 mission and the improved combined filter system having both nitrifying and denitrifying activities, and discuss its application to space experiments.

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