On April of 2000 and February of 2001, a disease characterized by darkening, ascites and enlargement of the spleen occurred in flounder, Paralichthys olivaceus, farmed near northern and eastern coasts of Korea. Various degree of necrosis including liquefactive-type was observed mainly in hemopoietic tissue of spleen and kindney. Certain tissue such as branchial soft tissue including epithelium showed a similar necrotic findings. But some tissues such as intentine had several necrotic clusters of wandering cells, being possibly leukocytes. Necrotic evidences of blood cells were evident in various tissues including hemopoietic tissues. Necrobiotic bodies were frequently encountered in blood smear preparations. Bullet type-virious were detected in the spleenic cells. Each virion was about 500 nm in diameter and 170nm in length. From these results, this disease was thought to be a rhabdoviral disease.

The objective of the study was to clarify the mechanism of persistent infection of marine birnavirus (MABV) in various nonpermissive cell lines. It was observed in CHSE-214, RTG-2 and RSBK-2 that the virus produced at high yield with typical cytopathic effect (CPE). On the contrary, the CPE was not produced in EPC, FHM and BF-2 cells. However amount of virus protein in both permissive and nonpermissive cell lines detected by ELISA was almost the same. Electron microscopy showed virions in permissive cells but not in nonpermissive cells. From the results, it is clear that virus protein and RNA were produced in nonpermissive cells as observed in permissive cells; however, assembly of the virus particles did not occur in nonpermissive cells.

The charcoal effects on the immune responses of flounder, Paralichthys olivaceus were studied. Fish were fed an experimental diet supplemented with 1%, 3% or 5% of charcoal to a commercial diet. Several factors such as weight gain, hematological parameters, nonspecific immune responses and resistance against Edwardsiella tarda were evaluated 4 and 8 weeks after the administraion. The changes of body weight and lysozyme activities were not signigicant among tested groups. But cholesterol and total protein concentration in the blood were significantly lower in the 5% supplemented group than in the control. The production of superoxide by macrophage from the head kidney, examined by NBT reduction, was significantly higher in the 3% charcoal supplemented groups than in the control. The survival rate after an artificial challenge with 1×10^exp(7) cells of E. tarda per fish was significantly the highest in the 3% charcoal supplemented group.

Effects of medicinal herb extract on nonspecific immune responses, hematology and disease resistance against Edwardsiella tarda in olive flounder, Paralichthys olivaceus were evaluated. Wormwood, Artemisia asiatica NAKAI and barrenwort, Epimedium koreanum NAKAI were mixed at a ratio of 7 : 3 (w/w) for 2-herbs extract and wormwood, barrenwort, Korean forsythia, Forsythia koreana NAKAI, chrysanthemum, Chrysanthemum zawadskii var. latilobum KITAMURA, peppermint, Mentha arvensis L. var, piperascens MALINV., great burnet, Snaguisorba afficinalis L., Lizard tail. Saururus chinensis BAILL., mulberry, Morus alba L., and star anise, Illicium varum HOOK, f, at the same weight for 9-herbs extract. Two-herbs of 9-herbs extract were prepared by heating after adding 10㎖ of distilled water per g of the herb mixtures. Fish (10.3$\pm$2.5g) were fed the experimental diets supplemented with the 2-herbs or 9-herbs extract at the different concentrations of 0%, 0.1%, 0.5% and 1% per kg diet for 12 weeks. Lysozyme and bactericidal activities of serum, and hematological characteristics were examined during experimental period. After feeding test period, all experimental groups were challenged with E. tarda. Lysozyme activity from the fish fed the diet supplemented with 0.1% or 0.5% of 2-herbs extract was significantly higher than the control. But there was no difference both in bactericidal activity and hematology among each group. Sixty seven % of relative percent survival values (RPS) in the group fed the diet supplemented with 0.1% of 2-herbs was higher than the other group and the control. These results suggest that supplenmentation of 0.1% of 2-herbs extract to a commercial diet may enhance disease resistance in olive flounder. Although both 0.1% and 0.5% 9-herbs extract did not improve non-specific immune reponses, they could enhance disease resistance of 53% RPS, respectively.

Effect of mercury (Hg) toxicity on survival, growth, feed efficiency and oxygen consumption were examined in the juvenile olive flounder. Paralichthys olivaceus. Fishes were exposed to sublethal concentrations of Hg ranging from 0 to 0.13mg/L for 6 weeks. Hg reduced survival rate in a concentration and exposure period-dependent way and suddenly reduction occurred at Hg concentrations greater than 0.05mg/L after 6 weeks. Growth rate and feed efficiency also significantly decreased at greater than 0.028 and 0.05 mg/L respectively. Oxygen consumption rate was significantly decreased to 25 and 32% than that of the control at the Hg concentration of 0.05 and 0.13 mg/L respectively. These results suggest that Hg toxicity inhibit physiological function including growth, feed efficiency and oxygen consumption in the juvenile olive flounder, resulting in survival failure at high concentration.

A blood biochemical analysis was carried out for the serum of wintering sea bream (red sea bream Pagrus major, black sea bream Acanthopagrus schlegeli, rock bream Oplegenathus fasciantuas) with green liver syndrome. GOT and GPT were significantaly elavated in the diseased sea bream. The value of $NH_{3}$ was slightly higher but those of TBIL. TP and UA were lower than the control. But there were no differences in albumin. Ca and glucose between the diseased and the control. These results suggent that a green liver syndrome in the sea bream might be related with nutritional defects and the stress of low water temperature during the winter.