蝦類養殖係為台灣傳統產業之一，並富有高度經濟價值。但由於
過度發展而不時有蝦類相關的疾病爆發，因此如何維繫養蝦業的發展
和永續性經營，除了養殖生物安全性應高度要求外，蝦體優質健康狀
態之維持亦為重點之一，因而深入了解其免疫作用之調控為目前重要
研究方向之一。
當蝦體遭受到細菌、病毒、真菌、寄生蟲感染或者遭受緊迫的情
況下，會產生自體免疫反應。哈威弧菌(Vibrio harveyi)為蝦類主要細
菌性病原體，其被證實在蝦類養殖過程當中，可造成嚴重感染狀況，
導致造成嚴重經濟損失。有鑑於免疫系統於抵抗病原入侵所扮演的防
禦機制，因此本研究則針對於哈威弧菌感染之免疫機制進行探討。本
研究方法目的即為利用quantitative real-time PCR 分析白蝦免疫相關
基因表現量的方式，以探討白蝦在遭受哈威弧菌感染後，其免疫系統
調控機制。本研究中所觀測之白蝦免疫系統的相關基因涵蓋了蝦類數
個免疫路徑，受檢體如下- 1. proPO 活化系統: Serine protease
(SerP)、proPO 。2.凝結系統: Transglutaminase (TGase)、clotting
protein (CP)。3. 抗菌胜肽系統Anti-LPS factor (ALF)、Lysozyme
(Lyz)、Penaeidin2 (PEN2)、Penaeidin3 (PEN3)、Penaeidin4 (PEN4)、
Crustin 。4. 抗氧化防禦系統: Superoxidase dismutase (SOD) 、

Glutathione peroxidase (GPx)。5. Pattern recognition receptor: Toll
receptor (LvToll)。
由目前實驗結果得知，白蝦免疫基因如LvToll、TGase、SOD、
proPO、Lyz、Crustin、ALF、PEN3、PEN4 等，在遭受哈威弧菌Vibrio
harveyi 感染後24 及36 小時，免疫基因表現量變化皆與控制組有顯
著差異，其中LvToll、Pen4 的表現量則達10~20 倍之差異性。
有鑑於Toll receptor 參與了無脊椎動物 (如:果蠅、蚊子…等)
對革蘭式陰性菌之免疫反應，因此本研究則進一步以蝦體內基因沉默
模式進行其於遭受哈威弧菌感染中扮演之角色分析。由實驗結果證
實，在Toll 基因沉默化後，白蝦對於哈威弧菌的感受性上升，除了在
感染後細菌清除能力下降之外，亦造成感染後死亡率有明顯上升的趨
勢，因而了解LvToll 於白蝦防禦系統當中，確實參與了革蘭式陰性菌
的免疫反應。

In this study, we used real-time PCR to simultaneously monitor the responses of 13
key genes of the shrimp innate immune system in Litopenaeus vannamei after
challenge with Vibrio harveyi. In the proPO activating system, we found that proPO
was upregulated (3.3  at 36 hpi) even though serine protease (SerP) was not. The
hemolymph cloting genes transglutaminae (TGase) and clotting protein were also
upregulated, as were 5 genes in antimicrobial peptide system (ALF, Crustin, Lyz, PEN
2 and PEN 4), with only PEN 3 showing no significant changes. In the antioxidant
defense system, SOD was slightly elevated while GPx was substantially
down-regulated. By contrast, in the pattern recognition receptor system, the Toll
gene (LvToll) showed both the highest (15.17 ) increases in expression levels t 24 hpi,
and this gene was therefore studied in more detail. When LvToll was knocked down
by RNAi silencing, there was no effect on either survival rates or bacterial number in
unchallenged shrimp. There was also no difference in mortality rates between control
shrimp and LvToll-silenced shrimp when these two groups were challenged with a
viral pathogen (white spot syndrome virus; WSSV). However, when LvToll -silenced
shrimp were challenged by Vibrio harveyi, there was a significant increase in
mortality and bacterial CFU counts. We note that the increase in bacterial CFU count
occurred even though treatment with EGFP dsRNA had the opposite effect of

reducing the CFU counts. We conclude that LvToll is an important factor in the
shrimp innate immune response to acute V. harveyi infection but not to WSSV.

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