Flounder Immune System And Resistance To Disease

In this thesis the immunity of one-year-old flounder, Paralichthys olivaceus, was studied in order to understand the mechanism of its resistance to diseases and to develop quick methods to measure the physical conditions of the fish. As the vegetative state is very important to the ability of the fish to resist diseases, the specific activities of certain digestive enzyme were studied too. The effects of recombinant yeast, in which salmon growth hormone gene is transferred, to the growth and non-specific immune parameters of Paralichthys olivuceus were studied In addition, the eDNA sequence of hepatic heat shock protein hsp7O was synthesized and amplified with PCR. This thesis consists of four major parts which include ten experiments. In experiment 1, the fine structure of the peripheral blood cells of paralichthys olivaceus was studied by light microscopy. Examination of Giemsa抯 stained smears of the blood revealed the blood cells include erythrocyte. thrombocyte, lymphocyte and monocyte. Though the erythrocyte and leucocyte can be classified easily, it is difficult to classify the leucocyte further. In experiment 2, electron microscopy examination of the ultrastructure of the peripheral blood cells of Parallchthys olivaceus showed that the blood cells were classifiable as erythrocytes(red cells)and leucocytes(white blood cells). The leucocytes were classifiable as lymphocytes, monocytes, thrombocytes and two types of granulocytes. The phagocyte-like bllod cell was observed too. According to the structure, no plasma cell was found in the peripheral blood. In experiment 3, the serum protein components of Paralichthys olivaceus were analyzed by capillary zone electrophoresis (CZE). Three kinds of proteins can be analyzed with current experimental conditions. Based on the time that these proteins appeared, they were named as P 2.59, P 1.60 and P 0.77. After storage at room temperature (25C) for 24 hours, the protein P 2.59 remained stable , while the other two proteins were destroyed. Comparison of the results with those of the analysis of human being抯 serum , the protein P 3 L 2.59 is probably attributable to Albumin, the protein P 1.60 is probably attributable to Globulin- a , the protein P 0.77 is probably attributable to Globulin- P. In experiment 4, the specific activities of antiprotease in some tissues of pseudoniale Paralichthys olivaceus were studied in this experiment. The specific activities in gill, mucus, serum and muscle were 2.56?.79, 6.81?.95, 5.18?.35, 3.64?.07 (unit/mm mg protein). From the results it can be concluded that the antiprotease exists in these tissues, which is important for the fish to protect themselves from the infection of the bacteria in the environment. In experiment 5, the specific activities of lysozyme in different tissues of pseudomale Paralichthys olivaceus were studied. The specific activities of lysozyme in mucus, serum, kidney, spleen were 0.58?.13,6.54?.19, 5.87?.97, 4.54?1.75, 2.33 ?.98(x10~3 unit/mining protein) respectively. The specific activities of lysozyme in liver, stomach, foregut, midgut, hindgut~ bile were 6.49 ?.35, 1.57?.65, 4.32?1.46, 2.07? 1.13, 2.41 ?1.29, 0.58 ?0.l3(xl0~3 unit/mm mg protein). The specific activity of lysozyme in gills was 3.96 ?1 .47(x 1 o~ unit/mm mg protein). No lysozyme activity w...