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論文名稱: | 低溫與氯化銨緊迫對吳郭魚血液之流變性、脂氧合酶及環氧合酶產物之影響
| The effect of low temperature and ammonium on blood characteristics, lipoxygenase and cyclooxygenase products in hybrid tilapia
研究生: | 劉德樂 De-Le Liu
指導教授: | 孫寶年 Bonnie Sun Pan
學位類別: | 碩士
學校名稱: | 國立台灣海洋大學
系所名稱: | 食品科學系
畢業學年度: | 91
語文別: | 中文
關鍵字: | 低溫緊迫 low temperature
| 氯化銨緊迫 stress
| 脂氧合酶 ammonium
| 血液流變性 blood viscosity
| lipoxygenase
全文點閱次數: | 11

全文影像連結:

電子檔1
電子檔2

論文頁數: | 90
摘要: | 中文摘要
|
| 雜交種紅色吳郭魚(全長約20.85 ± 1.21 cm/尾，重約157.37 ± 31.86 g/尾)在24℃馴養時，紅血球數約1.64 ± 0.31 ~1.74 ± 0.02 ×106 cell /mm3、白血球數目為5.47 ± 0.61 ~6.69 ± 0.32 ×104 cell/mm3、血比容積約24.63 ± 3.40 %、血糖濃度約0.40 ± 0.03 ~ 0.46 ± 0.13 mg /mL；吳郭魚曝露在以2℃/h的速率由24℃降溫11 ± 1℃後，維持12小時，紅血球數目增加2.20倍，血比容積變化與紅血球數相同、白血球數增加1.78倍、血糖含量則增加1.60倍，並造成行為改變；曝露24小時後，紅血球數則下降至2.02 ± 0.25 ×106 cell/mm3，血比容積也下降的現象，血糖持續增高至控制組3.65倍（p < 0.05），血球形態則無明顯變化。另以0.42℃/h之速率由25℃降至16 ± 1℃，維持72小時，在16 ± 1℃時，白血球數目由6.69 ± 0.32 ×104 cell/mm3增多1.52倍；在16 ± 1℃經24小時，紅血球數目由1.74 ± 0.02 ×106 cell / mm3增高1.24倍，但血色素、血比容積及血球形態則無明顯變化，血糖由0.40 ± 0.03 mg / mL增高至0.55 ± 0.07 mg / mL；在16 ± 1℃超過24小時後，血糖隨時間增加而下降，與控制組相同無顯著差異（p> 0.05）。
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| 根據σ = K γ n +C方程式觀察吳郭魚受低溫緊迫時對紅血球之流變性之影響，24℃時，其流體行為指標（flow behavior index；n）為0.95，黏稠度（flow consistence; K）為5.0 cP，在以2℃/h的速率降溫11 ± 1℃，紅血球n值升高至1.47 ~ 1.71 (r2 = 0.94)趨近於膨脹型流體，K值下降至0.47 cP。紅血球在切變速率0~ 200 sec-1，5分鐘，血比容積下降，並隨吳郭魚受低溫緊迫時間的延長下降比例增加。在16 ± 1℃經過48小時，n值上升至1.60，血漿中前列凝素(11-dyhydro thromboxane B2) 含量自0.085 ng/ mL增加至0.67 ng/ mL，顯示受到低溫刺激血液中血小板活化；超過48小時後，n值下降至1.22。
|
| 在急速降溫實驗經18小時後，以化學冷光法測得吳郭魚血液之脂氧合酶(lipoxygenase; LOX)活性由330.6 ± 20.9 mV / mg protein-min增加至447.12 ± 25.4 mV / mg protein-min（p < 0.05）。血漿中白三烯素（Leukotriene B4; LTB4）含量由11.7 pg/mL下降至8.3 pg/ mL。至30小時後，活性增加為控制組之3.46倍（p < 0.05），白三烯素含量增高為2.01倍，顯示此時血液組織中5-LOX活性有明顯增高；在緩慢降溫至16 ± 1℃時，其LOX活性增高2.20倍，持續超過48小時後，活性下降，與控制組無顯著差異（p < 0.05），變化之趨勢較血糖變化之趨勢相同顯示吳郭魚已適應16 ± 1℃之環境。
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| 吳郭魚曝露在3.0 mg/L氯化銨之水中超過24小時，紅血球數由1.80 ± 0.45 ×106 /mm3減少至1.68 ± 0.44 ×106 /mm3，而白血球數則由17.57± 1.52 ×104 cell/mm3隨濃度增高至23.26 ± 32.15 ×104 cell/mm3；LOX在3.0 mg/L氯化銨之水中超過24小時，活性由539 ± 24 mV / mg protein-min增高至1920 ± 62 mV / mg protein-min；紅血球明顯縮小(p < 0.05)。血糖、血比容積則無明顯之變化；在含1.5 mg/L氯化銨之水中超過72小時，紅血球降至2.07 ± 0.22 ×106 /mm3與控制組無顯著差異、白血球數則隨時間延長而增加、血糖含量則隨曝露時間的延長增高1.70倍，在含1.5 mg/L氯化銨之水中超過72小時，LOX活性增高2.86倍。
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| 血液中LOX活性變化較紅血球數目、白血球數目、血比容積、血糖更能快速且靈敏反應出吳郭魚受緊迫之程度。
|
| Abstract
|
| Red tilapia hybrid (total length 20.85 ± 1.21 cm, total weight 157.37 ± 31.86 g) preadapted at 24℃ had erythrocyte and leukocyte counts of 1.64 ± 0.31 ~1.74 ± 0.02 ×106 cell3 /mm and 5.47 ± 0.61 ~ 6.69 ± 0.32 ×104 cell/mm3 respectively, a hematocrit of 24.63 ± 3.40 % and plasma glucose of 0.40 ± 0.03 ~ 0.46 ± 0.13 mg /mL. Tilapia exposed to temperature reduced at a rate of 2℃/h to 11 ± 1℃ then maintained for 12 h, showed behavioral change, increased plasma glucose to 1.6 times, increased erythrocyte and leukocyte counts to 2.20 and 1.78 time of the control (p<0.05); after exposure for 24 h, erythrocyte counts decreased to 2.02 ± 0.25 × 106 cells/mm3, hematocrit also decreased, the plasma glucose increased to 3.65 times of the control (p<0.05). Tilapias exposure to temperature reduced at 0.42℃/h from 25℃to 16 ± 1℃and maintained for 72 h, at 16 ± 1℃, leukocyte counts increased by 1.52 time of the control; at 16 ± 1℃ through 24 hours, erythrocyte counts and plasma glucose increased to 1.24 and 1.37 times of the control while hemoglobin, hematocrit and erythrocyte shape showed no significant difference from those of the control., the plasma glucose level decreased afterwards to the level of the control (p>0.05).
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| Blood thinning effects were observed in fish exposed to rapidly reduced temperature. The rheological behavior fitted the power law equation σ = Kγ n + C. At 24℃. Flow behavior index (n) was 0.95, while flow consistency (K) was 5.0 cP. As the temperature reduced at 2℃/h to 11 ± 1℃, erythrocyte flow behavior index increased with the exposure time of tilapia at 11 ± 1℃ to 1.47 ~ 1.71 (r2 = 0.94) being closer to dilatant-flow. Flow consistency decreased to 0.47 cP. Shearing at a rate from 0 to 200 secs-1 for 5 minutes, the hematocrit decreased with prolonged exposure time. As temperature reduced to 16 ± 1 ℃ and maintained for ≧ 48 h, the n value went up to 1.60, then decreased to 1.22 afterwards, plasma 11- dehydro thromboxane B2 increased from 0.085 ng/ mL to 0.67 ng / mL indicative of platelet activation by low temperature.
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| The tilapia exposed to rapid reduction in temperature showed increases in blood lipoxygenase (LOX) activity measured as chemiluminescence intensity. If increase from 330 □ 20.9 mV/mg protein-min to 447.12 □ 25.4 mV/mg protein-min (p<0.05). Plasma leukotriene B4 (LTB4) concentration decreased from 11.7 pg/mL to 8.3 pg/mL. After 30 h at low temperature, LOX activity increased to 3.46 times and LTB4 concentration increased to 2.01 times of the control (p<0.05). During gradual reduction of temperature to 16 ± 1 ℃, blood LOX activity was 2.20 times of the control, after 48 h, LOX activity did not change significantly from that of the control (p<0.05), being a sign that tilapia had adapted to the low temperature of 16 ± 1 ℃.
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| Tilapia exposed to 3.0 mg/L ammonium chloride for over 24 h, erythrocyte count decreased from 1.80 0 ± 0.45 ×106 /mm3 to 1.68 ± 0.44 ×106 /mm3, leukocyte count increased from 17.57± 1.52 ×104 cell/mm3 to 23.26 ± 32.15 ×104 cell/mm3, plasma LOX activity increased from 539 ± 24 mV / mg protein-min to 1920 ± 62 mV / mg protein-min. Erythrocyte decreased in size (p<0.05). Plasma glucose and hematocrit showed no significant differences from the control (p>0.05). Tilapia exposed to 1.5 mg/L of ammonium chloride for 72 h, the erythrocyte count was not significantly different from the control (p>0.05), leukocyte count increased with prolonged exposure time, plasma glucose increased to 1.70 times, while LOX activity increased to 2.68 times of the control.
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| The tilapia blood LOX activity seemed to respond with more sensitivity to the physical and chemical environmental stresses than erythrocyte count, leukocyte count, hematocrit, hemoglobin and plasma glucose.
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論文目次: | 目錄
| 目錄    Ⅰ
| 圖目錄    Ⅳ
| 縮寫表    Ⅵ
| 中文摘要    Ⅶ
| 英文摘要    Ⅸ
|
| 壹、前言    1
|
| 貳、文獻回顧    3
| 一、緊迫因子對於魚類的影響    3
| (一) 魚類之緊迫反應    3
| (二) 溫度變化之影響    4
| (三) 氨氮化合物之影響    6
| 二、魚類的血液    8
| (一) 一般組成    8
| (二) 血液的流體性質    9
| 1. 剪切速率的影響    10
| 2. 血比容積    10
| 3. 血液的凝集    11
| 三、環境對魚類血液的影響    11
| 四、血液的脂氧合酶    13
| 五、環氧合酶對於血液的影響    16
|
| 參、材料與方法    18
| 一、吳郭魚    18
| 二、方法    18
| (一) 魚體測定    18
| (二) 低溫緊迫實驗    18
| 1. 急速降溫實驗    18
| 2. 緩慢降溫實驗    19
| (三) 氨化銨毒性實驗    19
| 1. 氯化銨毒性緊迫實驗    19
| 2. 氯化銨毒性緊迫適應實驗    19
| (四) 血液分析    20
| 1. 抽血    20
| 2. 血液基本參數值    20
| （1）紅血球個數    20
| （2）白血球個數    20
| （3）血色素    21
| （4）血比容積    21
| 3. 血液滲透壓之測定    21
| 4. 血液流變性質    21
| 5. 抹片染色    22
| 6. 紅血球滲透脆性試驗    22
| 7. 血糖測定    23
| (五) 血液中脂氧合酶之測定    24
| 1. 樣品製備    24
| 2. 粗酵素製備    24
| 3. 冷光測定法    25
| 4. 吸光法    26
| 5. 高效液相層析法    26
| (六) 血液中eicosanoid類產物測定    27
| 1. 白三烯素之含量    28
| 2. 前列凝素之含量    28
| (七) 蛋白質定量    28
|
| 肆、結果與討論    30
| 一、低溫緊迫因子對吳郭魚行為之影響    30
| 二、低溫緊迫對血糖之影響    32
| 三、低溫緊迫對吳郭魚血球之影響    37
| (一) 血球數目    37
| 1. 紅血球數    37
| 2. 白血球數    42
| (二) 紅血球外型之改變    45
| (三) 紅血球細胞膜強度之改變    48
| 四、紅血球流變性    49
| 五、血漿中脂氧合酶活性及白三烯素含量之變化    59
| 六、吳郭魚對氯化銨造成之環境緊迫與低溫緊迫之差異    64
|
| 伍、結論    78
|
| 陸、參考文獻    80
|
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2009 年 11 月 22 日 23:47