本論文使用由Schizophyllum commune純化之多醣類(b-1,3-glucan, BG)，針對各體型草蝦進行一系列研究，探討BG增強草蝦免疫能力之機制，並定量BG使用後對增進免疫能力的效能。研究分為三大部分；第一部份探討口投多醣後，對草蝦稚蝦生長的影響及對弧菌感染的抵抗能力。由成長試驗之結果發現，連續18週餵飼含多醣(0、0.2、2、10 g/kg diet)飼料，對於草蝦稚蝦(0.5 ± 0.1 g)之生長並無不良影響(P>0.05)。而經感染試驗結果，多醣添加量至少應為2 g/kg diet，並且需連續餵飼10天以上，才能顯著(P<0.001)增強草蝦(20.4 ± 2.1 g)抵抗弧菌之感染。將多醣進一步應用於強化草蝦苗活力之效果發現，餵飼添加多醣2 g/kg diet之飼料可明顯(P<0.001)強化草蝦苗耐飢餓與抗V. harveyi的感染。而以多聚磷酸態維生素C (PAA 0, 0.2, 1.0 g/kg)配合多醣(2 g/kg diet)共同餵飼草蝦(8.2 ± 1.5 g) 20天，只有添加多醣組，在餵飼10天後可較顯著之抗Vibrio damsela感染(P<0.0001)。經20天餵飼後，則以添加BG 2 g/kg diet + PAA 0.2 g/kg diet組之活存率60 %為最高，具有明顯之加成抗菌效果(P<0.0001)。
第二大部分在比較口投含多醣飼料，對草蝦受傷後之組織復原能力與增強草蝦種蝦體內免疫功能之效果。在餵飼PAA 0, 0.2, 1 g/kg diet與BG 2 g/kg diet等四種飼料30天後，經人為割傷之草蝦(8.2 ± 1.5 g)組織復原能力，以添加PAA 1.0 g/kg diet組為最快。而多醣添加可降低草蝦傷口組織受細菌感染的機會，但對組織之復原幫助不大。草蝦種蝦(135 ± 25 g)餵飼含多醣(0, 2 g/kg diet)飼料40天後，餵飼添加多醣，蓄養於室內與室外之種蝦活存率(71.7與70 %)，顯著(p<0.001)較未添加組(34.5與24.3 %)高。攝食含多醣飼料之種蝦，其血淋巴球之附著、吞噬能力與Superoxide anion產生量(P<0.001)均優於未添加者。
第三大部分探討口投含多醣類飼料，對增強草蝦抵抗白點症病毒(WSSV)之效果。蝦苗(PL15)與稚蝦(5.5 ± 0.5 g)經餵飼添加多醣(0, 2 g/kg diet)飼料15與20天後以WSSV感染，結果顯示，兩種大小草蝦活存率分別為12.2與20 %，顯著(P<0.005)優於未添加組的0 %。進一步增加多醣之添加劑量時，口投含多醣10 g/kg diet組在感染WSSV後有42 %活存，優於(P<0.0001)其他各組(0, 1, 2, 20 g/kg diet)。試驗蝦感染WSSV後，蝦體內之血淋巴球數目、血淋巴球吞噬能力、酚氧化

Three series of studies were conducted to quantify the effectiveness of dietary incorporation of b-1,3-glucan (BG) from Schizophyllum commune in enhancing the immunity and resistance of grass prawn Penaeus monodon to vibriosis and white spot syndrome virus (WSSV) infections.
In the first series of studies, three experiments were conducted to evaluate the effectiveness of dietary b-1,3-glucan on shrimp growth and resistance to vibriosis. Weight gain, survival and feed efficiency of juvenile shrimp (0.5 ± 0.1 g) were not significantly different (P>0.05) after being fed the diets containing BG 0, 0.2, 2, 10 g/kg diet for 18 weeks. Subadult shrimp (20.4 ± 2.1 g) fed the diet containing of BG 2 g/kg diet for 10 days showed a significantly (P<0.001) enhanced resistance against vibriosis. Postlarvae fed with the BG diet (2 g/kg diet) were more resistant (P<0.001) against starvation and V. harveyi challenges than the postlarvae fed non-BG diet. Additive disease resistance was observed when polyphosphorylated L-ascorbic acid (PAA) was used together with BG. In challenge tests with V. damsela, shrimp fed with PAA (0.2 g/kg diet) + BG (2 g/kg diet) diet for 20 days had a survival rate up to 60%.
In the second series of studies, two experiments were conducted to evaluate the effectiveness of dietary b-1,3-glucan on wound healing and immunity in spawners. Dietary supplement of BG reduced the chance of infections, but did not help wound healing as did the supplement of PAA. And regardless of indoor or outdoor rearing, the survival rate of brooder (135 ± 25 g) fed the BG (2 g/kg diet) diet was higher (P<0.001) than that of the non-BG group. Fed the BG brooders showed enhanced haemocyte phagocytic activity, cell adhesion and superoxide anion production then the control group.
Third series of studies evaluated the effectiveness against white spot syndrome virus (WSSV). Six days after being challenged with WSSV, 12.2 % of the BG-treated (2 g/kg diet for 15 days) postlarvae (PL15) and 20 % BG-treated (2 g/kg diet for 20 days) juveniles (5.5 ± 0.5 g) were still alive; while all non-BG-treated shrimp died. In order to quantify the effectiveness of BG to WSSV, juveniles (6.5 ± 0.4 g) were fed diets containing graded levels of BG. The results showed that shrimp fed the diet containing BG 10 g/kg for 20 days had the highest (P<0.001) survival rate (42 %) among all groups. Shrimp that received diets supplemented with BG at a dosage >2 g/kg recuperated 9 ~ 12 days after WSSV challenge; while the group fed diets with no or 1 g/kg BG suffered from rapid decrease in total haemocyte count, phagocytic activity, phenoloxidase, O2-, superoxide dismutase (SOD) production and subsequent high mortality.
The results in this study showed that b-1,3-glucan is effective in enhancing the phagocytic activity, phenoloxidase, O2- and SOD productions and consequently the resistance of postlarval, juvenile, subadult and brooder P. monodon against vibriosis and viral infections. Since prolonged use of BG, even at optimal dietary levels, decreased the immunity of the shrimp, care therefore must be taken to maximize its effectiveness. A cycle of dietary BG supplement of 2 ~ 10 g/kg diet for 20 days with an intermission of 10 days may serve the purposes.

目 錄

中文摘要 I
英文摘要 III
目錄 V
表目錄 VI
圖目錄 VIII
相片目錄 IX
第一章、緒言 1
第二章、綜合材料與方法 7
第三章、多醣類對草蝦抵抗弧菌感染之效果 19
3-1、多醣類促進草蝦成長與抗弧菌感染能力 20
3-2、多醣類應用於強化草蝦幼苗抗飢餓與抗弧菌感染能力 30
3-3、多聚磷酸態維生素C與多醣類的共同效應 37
第四章、多醣類對草蝦傷口復原與免疫能力的影響 46
4-1、多醣類與受傷之傷口復原能力 47
4-2、多醣類促進草蝦種蝦免疫力與生殖表現 56
第五章、多醣類對強化草蝦抵抗白點症病毒(WSSV)之效果 68
5-1、多醣類增強稚蝦與蝦苗抵抗白點症病毒能力 69
5-2、提高多醣類使用量可強化草蝦抵抗白點症病毒的能力 83
第六章、結論及建議 108
第七章、參考文獻 111
附錄一、內文簡寫表 126
附錄二、本論文研究期間發表之相關文章以ATP分析儀定量草蝦血淋巴球之總數量 128

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