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研究生:李之琴
研究生(外文):Putri Nurhanida Rizky
論文名稱:評估黄芩(Scutellaria baicalensis)熱水萃取物抗海鱺(Rachycentron canadum)發光菌(Photobacterium damselae subsp. piscicida)與寄生蟲(Piscicola geometra)之作用
論文名稱(外文):Evaluation of Scutellaria baicalensis root hot water extract effects on Photobacterium damselae subsp. piscicida and Piscicola geometra of cobia (Rachycentron canadum)
指導教授:鄭達智 博士
指導教授(外文):Dr. Ta Chih Cheng
口試委員:邱亞伯 博士 鄭達智 博士
口試委員(外文):Dr. Ir. Happy Nursyam, MSDr. Albert Linton CharlesDr. Ta Chih Cheng
口試日期:2017-05-04
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:熱帶農業暨國際合作系
學門:農業科學學門
學類:一般農業學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:英文
論文頁數:89
中文關鍵詞:黃芩根部海鱺餵食先天性免疫浸浴
外文關鍵詞:Scutellaria baicalensis rootcobiaoral feedinginnate immunebathing
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黃芩之根部已被列為可用藥物作為抗炎、抗病毒、抗癌、自由基消除劑、抗氧化劑和免疫刺激之作用。黃芩之根部含有高含量之黃酮類化合物(高於40%),包括黃芩苷、漢黃芩素、漢黃芩苷、白楊素、木蝴蝶素甲、黃芩黃酮、及其他等。本研究將利用水萃取物之方法來提煉黃芩之根部以不同之濃度(0%、0.5%、1%、2%、3%、及5%)來加進去海鱺之魚飼料,分析抗菌和抗寄生蟲之反應。結果發現1%濃度可以抗發光桿菌及抗寄生蟲感染,並能提高飼料利用率。同時增強了血清中之過氧化酶、過氧化物酶、和溶菌酶。在基因表現中,黃芩根部萃取物增加脾臟中之Toll受樣體、介白素-1β、免疫球蛋白M、及腫瘤壞死因子-α基因表現。不同稀釋程度之黃芩根部提取物之毒性顯示出,將海鱺放置於500X和1000X稀釋後之提取液中達72小時後,高達60%之海鱺身體表面至底部的寄生蟲(尺蠖魚蛭)會脫離魚身或死亡。此外,解剖後的海鱺沒有明顯的魚鰓、脾、腎臟和肝臟的損傷。這項研究證明黃芩根部提取物可以是海鱺免疫增強劑。
關鍵詞:黃芩根部、海鱺、餵食、先天性免疫、浸浴
The root of Scutellaria baicalensis has been used as a drug for anti-inflammatory, antiviral, anticarcinogenic, free-radical scavenging, antioxidant and immunostimulatory effects. The root of S.baicalensis has a high content of flavonoids (over 40%) including baicalin, wogonin, wogonoside, chrysin, oroxylin-A, and skullcap flavones. The aim of this study is to analyze the antibacterial and anti-parasitic activity of S.baicalensis root by water extraction, supplemented in cobia fed and as a bath treatment. S.baicalensis root water extract supplemented in fish feed failed to reduce the number of marine leeches. However, it showed high accuracy to enhance the growth performance, feed utilization, and disease resistance against Photobacterium damselae subsp. piscicida after 30 days of feeding health cobia and 90 days with leech infestation. Cobia fed supplemented S.baicalensis root water extract also showed significant enhancement of myeloperoxidase content, peroxidase content, and lysozyme activity in cobia serum. Moreover, S.baicalensis root water extract up-regulated the expression of TLR-9, IL-1β, IgM, and TNF-α in the spleen. 1% concentration of S.baicalensis root water extract supplemented in feed showed the high accuracy to enhance growth performance, innate immune system and disease resistance of cobia against bacteria infection. Bathing treatment of cobia shows the 60% mortality and detachment of the leeches (Piscicola geometra) after 72 hours exposure with 500X and 1000X dilution of S.baicalensis root water extract. Therefore, there was no significant cell damage, or lesions of the cobia gills, spleen, kidney, and liver. This study proved the use of S.baicalensis root water extract as an immunostimulant in cobia culture industry with significant efficacy, low cost, and ease of practice.
Keywords: Scutellaria baicalensis root, cobia, oral feeding, innate immune, bathing,
摘要 I
Abstract II
Acknowledgments IV
List of tables X
List of figures XI
1. Introduction 1
1.1 Background 1
1.2 Objectives 3
2. Literature review 4
2.1 Cobia (Rachycentron canadum) 4
2.1.1 Taxonomy and morphology of cobia 4
2.1.2 Distribution of cobia 5
2.1.3 Disease of cobia (Rachycentron canadum) 7
2.2 Scutellaria baicalensis 10
2.3 Fish immune organ 12
2.4 Fish immune system 13
2.4.1 Cytokine 15
2.4.2 Humoral defense factor 15
3. Materials and methods 20
3.1 Flow chart for feeding trial of cobia and immune responses studies 20
3.2 Experimental design feeding trial 20
3.3 Feeding trial and fish experimental design 21
3.4 Extract preparation and stock solution 22
3.4.1 Bacterial challenge 22
3.4.2 Growth rate 23
3.4.3 Fish blood and tissue collection 23
3.5 Humoral immunity 24
3.5.1 Myeloperoxidase content 24
3.5.2 Peroxidase activity 24
3.5.3 Serum lysozyme activity 24
3.5.4 Total RNA isolation and cDNA synthesis 25
3.5.5 qRT-PCR 26
3.6 Statistical analysis 27
3.7 Flow chart examination of toxicity of Scutellaria baicalensis root aqueous extract 28
3.8 Leeches experimental design 29
3.9 Fish experimental design of bathing treatment 29
3.10 Histology 30
3.11 Statistical analysis 30
4. Result and discussion 31
4.1 Effect of S.baicalensis root water extract on the growth performance, feeding rate, and survival rate of cobia (Rachycentron canadum) 31
4.1.1 Fish growth performance (1st experiment of feeding trial) 31
4.1.2 Fish growth performance (2nd experiment of feeding trial) 38
4.2 Effect of Scutellaria radix water extract on the non-specific immune responses of cobia (Rachycentron canadum) 45
4.2.1 Serum myeloperoxidase content 45
4.2.2 Serum peroxidase content 46
4.2.3 Serum lysozyme activity 47
4.3 Effect of S.baicalensis root water extract on the cytokine gene related of cobia (Rachycentron canadum) 50
4.4 Behavior of Piscicola geometra 56
4.5 Toxicity of S.baicalensis root water extract on Piscicola gemoetra in cobia culture 57
4.6 Toxicity of S.baicalensis root water extract on cobia (Rachycentron canadum) 59
4.6.1 Behavior of cobia (Rachycentron canadum) 60
4.6.2 Histology 61
4.6.3 Effect of leech in cobia culture 66
4.6.4 Effect of Toxicity of S.baicalensis root water extract on Piscicola geometra attack to the cobia body 67
5. Conclusion 70
References 72
Bio sketch of Author 88
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