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Histological survey was made to determine nitrate toxicity on the Medaka fish, Oryzias latipes. In order to investigate the effects of short-term exposure to nitrate, one-month-old Medaka fish was exposed to NaNO 3 at concentrations of 100 and 125 mg NO 3 -N l -1 for 96 hours. At the end of the exposure period, survival rate was found to be 30% and 10%, for the 100 and 125 mg NO 3 -N l -1 exposure concentrations, respectively. Histological examination of the organs showed that disruption of cell alignment was a common feature in the gills, intestinal ampulla, liver and kidney. A long-term exposure experiment was also carried out, whereby Medaka fish was exposed to NaNO 3 (100 and 125 mg NO 3 -N l -1 ) for three months from its egg stage. Eggs treated with NaNO 3 hatched within 10 days after fertilization. At the end of the exposure period, survival rate in the 100 and 125 mg NO 3 -N l -1 treatments were 40% and 30%, respectively. Fibrosis of the hepatic cells and curved spinal column were observed in the juveniles subjected to long-term nitrate exposure. The results of our experiments suggest that the high mortality resulting from short-term acute exposure to nitrate is caused by general dysfunction throughout the whole body. The chronic toxic effects attributed to nitrate, following long-term exposure, were likely to have resulted from nutrient deficiency caused by hepatic dysfunction.

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.