酪胺對淡水長臂大蝦生理及免疫反應之影響

Effects of tyramine on the physiological and immunological responses of giant freshwater prawn, Macrobrachium rosenbergii

劉昌奇(Liu Chang-Chi)

指導教授 ： 鄭文騰

國立屏東科技大學/農學院/水產養殖系所/碩士(2020年)

https://doi.org/10.6346/NPUST202000342

摘要

酪胺 (tyramine)為一種生物單胺類，於無脊椎動物中參與多種重要之生理作用，包括免疫調節。本研究目的為評估酪胺對淡水長臂大蝦免疫能力之影響。試驗分為兩部分包括酪胺注射與餵食試驗， (1)注射酪胺濃度為 0、1、10 nmol prawn-1，於0、0.5、1、2、4、8小時採樣， (2)飼料添加酪胺濃度為0、1、10 mg (kg diet)-1，餵食試驗持續7天，並於0、1、3、7天採樣，檢測其總血球數 (THC)、不同血球數 (DHC)、酚氧化酵素活性 (PO)、超氧陰離子活性 (O2-)與溶菌酶活性之免疫反應變化；血淋巴葡萄糖、乳酸、多巴胺 (DA)與正腎上腺素 (NE)之生理變化；對乳酸鏈球菌 (Lactococcus garvieae)之吞噬作用、清除效率以及攻擊試驗對病原體之抵抗性；測定免疫相關基因表現量。注射試驗結果顯示，酪胺注射1 nmol prawn-1試驗組於注射後0.5小時其總血球數、透明血球、半顆粒血球、酚氧化酵素活性及其注射後1小時對乳酸鏈球菌之吞噬活性、清除效率顯著增加。酪胺注射10 nmol prawn-1試驗組於注射後0.5小時其溶菌酶活性、吞噬活性、清除效率及血淋巴葡萄糖、多巴胺含量顯著降低。注射酪胺濃度為1、10 nmol prawn-1試驗組於0.5小時其呼吸爆與血淋巴乳酸顯著高於控制組，此外，酪胺注射10 nmol prawn-1試驗組於注射後0.5小時抑制多巴胺釋放且具有濃度關係。酪胺注射後2小時，所有免疫指標均恢復至生理值。酪胺注射1小時後，進行乳酸鏈球菌攻擊試驗，酪胺注射1 nmol prawn-1試驗組之活存率顯著提升；餵食試驗結果顯示，餵飼添加酪胺1、10 mg (kg diet)-1試驗組於第3天其總血球數、顆粒血球、半顆粒血球、酚氧化酵素活性及超氧陰離子活性顯著高於控制組。餵飼添加酪胺10 mg (kg diet)-1試驗組於第7天其超氧陰離子活性顯著高於控制組。餵飼添加酪胺1、10 mg (kg diet)-1試驗組於第3天其吞噬作用及清除效率顯著高於控制組。所有免疫指標皆於餵飼7天後恢復至生理值與控制組無顯著差異。淡水長臂大蝦餵飼含酪胺飼料亦顯著提升經乳酸鏈球菌攻擊後之活存率。此外，攻擊後持續餵食添加酪胺10 mg (kg diet)-1試驗組其存活率於168小時能夠維持90%。餵飼添加酪胺1 mg (kg diet)-1試驗組於第3天其proPO系統相關基因表現量顯著高於控制組。以上結果證明神經傳遞物質酪胺不論以注射或餵食方式，於短期間內皆可能促進免疫調節，且增強淡水長臂大蝦對乳酸鏈球菌之抵抗力。

關鍵字

淡水長臂大蝦 ； 酪胺 ； 免疫能力 ； 乳酸鏈球菌

並列摘要

Tyramine (TA), a biogenic monoamine, plays various important physiological roles and immunological regulation in invertebrates. The purpose of this study was to evaluate the effects of tyramine on the immune responses of giant freshwater prawn, Macrobrachium rosenbergii. Two experiments TA injection and feeding were conducted to examine the effects on physiological responses and immunological regulation. (1) In injection group, M. rosenbergii (9.2  0.5 g) received TA at 0, 1, 10 nmol prawn-1, and were sampled at 0, 0.5, 1, 2, 4, 8 hrs. (2) In feeding group, three experiment diets, including the control diet (without of tyramine), dietary supplement of TA 1 and 10 mg (kg diet)-1 were carried out. During 7 days of culture, M. rosenbergii (10.3  0.2 g) was sampled at 0, 1, 3, 7 days. The total hemocyte count (THC), different hemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (release of superoxide anion), lysozyme activity, Hemolymph glucose, lactate, dopamine and norepinephrine, phagocytic activity, clearance efficiency, challenge test to the pathogen Lactococcus garvieae and immune-related genes expression were measured. TA-injection results showed that THC, hyaline cells, semigranular cells, and PO activity at 0.5 hr as well as phagocytic activity and clearance efficiency to L. garvieae at 1 hr of prawn injected with TA at 1 nmol prawn-1 significantly increased, but the significantly decreased plasma lysozyme activity, phagocytic activity, clearance efficiency, and Hemolymph glucose and dopamine were observed in prawn injected with TA at 10 nmol prawn-1 for 0.5 hr. Respiratory bursts and Hemolymph lactate in two TA-injection treatments at 0.5 hr, were significantly higher than those of the saline control, and in addition, TA depressed dopamine release in a dose-dependent manner after 0.5 hr of TA 10 nmol prawn-1-injection. All the examined parameters returned to control levels after prawn injected with TA for 2 hrs. For prawn received TA for 1 hr then challenged with L. garvieae, the survival ratio of TA 1 nmol prawn-1-injected prawn significantly increased. Feeding results showed that THC, granular cells, semigranular cells, PO activity, respiratory bursts of prawn fed with dietary supplement of TA 1 and 10 mg (kg diet)-1 significantly increased at 3 days. In prawns fed with dietary supplement of TA 10 mg (kg diet)-1, respiratory bursts had increased at 7 days. Phagocytic activity and clearance efficiency of prawn fed with dietary supplement of TA 1 and 10 mg (kg diet)-1 significantly increased at 3 days. All of the immune parameters had returned to control values by 7 days after prawn fed. Dietary supplement of TA 10 mg (kg diet)-1 significantly increased the survival ratio (80%) of M. rosenbergii challenged with the pathogen L. garvieae after 72 hrs. However, continuous feeding dietary supplement of TA 1 and 10 mg (kg diet)-1 also significantly increased the survival ratio (73% and 90%) of M. rosenbergii challenged with L. garvieae after 168 hrs. proPO system-related genes expression of prawn fed with dietary supplement of TA 1 mg (kg diet)-1 significantly increased at 3 days. These results suggest that neurotransmitters TA to dietary supplement promotes the immune responses, which in turn enhance the resistance against to L. garvieae of M. rosenbergii.

並列關鍵字

Macrobrachium rosenbergii ； Tyramine ； Immunity ； Lactococcus garvieae

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