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研究生:陳達煒
研究生(外文):Ta-Wei Chen
論文名稱:靈芝固態發酵產物之抗氧化活性分析及其對離乳仔豬生長性狀和細胞激素表現的影響
論文名稱(外文):Analysis of antioxidant activities from Ganoderma lucidum solid-state fermentative products and its effect on growth performance and cytokine expression in weanling piglets
指導教授:鄭永祥鄭永祥引用關係陳淑德
指導教授(外文):Yeong-Hsiang ChengSu-Der Chen
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:生物技術研究所碩士班
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
中文關鍵詞:靈芝固態發酵離乳仔猪抗氧化細胞激素
外文關鍵詞:Ganodermasolid state fermentationweanling pigletantioxidantcytokine
相關次數:
  • 被引用被引用:2
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摘要
本研究為利用小麥等穀類基質、培養天數及水分含量等因素,進行靈芝固態發酵。以酚硫法分析靈芝固態發酵產物的多醣濃度及清除DPPH自由基能力、還原力和螯合亞鐵離子能力,以評估靈芝抗氧化活性。添加不同劑量(0 ppm、250 ppm、500 ppm、750 ppm和1000 ppm)靈芝固態發酵產物至飼料中,以評估其對離乳仔猪飼養生長表現、血液生化值及腸道黏膜DNA、RNA和protein含量及重要淋巴組織之細胞激素表現量的影響。結果顯示,靈芝固態發酵產物發酵一星期之多醣產量由多至少,依次為稻米、薏仁、小麥、玉米和大豆,由於稻米和薏仁基質會產生液化且菌絲不易生長,因此以小麥為主要培養基。大豆基質之固態發酵產物,則具有良好的抗氧化活性。250 ppm和750 ppm靈芝固態發酵產物的處理組對於離乳仔猪每日平均增重和飼料換肉率有較佳之生長性能現。由血液生化分析結果發現,在第二週抽血時,250 ppm的處理組具有降低血糖和三酸甘油酯的效果,而500 ppm和750 ppm的處理組在肝功能的指標上,仔猪血液中的ALT及AST值有明顯下降。在第四週以500 ppm的處理組最具上述之保健效果。腸道黏膜表現結果顯示,空腸和迴腸黏膜DNA、RNA在750ppm跟其他處理組做比較,具有顯著性較高的表現量。盲腸及十二指腸蛋白質在500 ppm較其他處理組表現量較高。在仔豬肺門淋巴、鼠蹊淋巴和脾臟偵測細胞激素IL-1β和IFN-γ在750 ppm分別增加50%及30% mRNA的表現量,IL-2、IL-4、TNF-α在1000 ppm分別增加50%、200%和40% mRNA的表現量,由結果推測靈芝發酵產物具有促進細胞性免疫的提升,故靈芝發酵產物添加於飼料中可調節離乳仔猪的免疫力。

關鍵詞:靈芝、固態發酵、離乳仔猪、抗氧化、細胞激素
總目錄
中文摘要----------------------------------------------------------------------------І
英文摘要--------------------------------------------------------------------------Ш

致謝-------------------------------------------------------------------------------- V

總目錄--------------------------------------------------------------------------- VII

表目錄--------------------------------------------------------------------------- ΧII

圖目錄-------------------------------------------------------------------------- ΧIII

壹、前言---------------------------------------------------------------------------1

貳、文獻回顧---------------------------------------------------------------------2

一、靈芝分類---------------------------------------------------------------------2

(一)靈芝特徵介紹----------------------------------------------------------------3

(二)靈芝生活史-------------------------------------------------------------------3

(三)靈芝生理活性功效----------------------------------------------------------5

1.降血壓----------------------------------------------------------------------------5

2.降血糖----------------------------------------------------------------------------5

3.降低膽固醇----------------------------------------------------------------------5

4.抗腫瘤----------------------------------------------------------------------------6

5.抗氧化----------------------------------------------------------------------------6

6.抗發炎----------------------------------------------------------------------------7

(四)靈芝的有效成分-------------------------------------------------------------8
二、靈芝液態發酵影響因子---------------------------------------------------10

(一)培養基成分------------------------------------------------------------------10

1.碳源----------------------------------------------------------------------------- 11

2.氮源----------------------------------------------------------------------------- 12

3.碳氮比-------------------------------------------------------------------------- 12

4.無機鹽類----------------------------------------------------------------------- 13

(二)培養條件---------------------------------------------------------------------13

1.溫度----------------------------------------------------------------------------- 13

2.pH值---------------------------------------------------------------------------- 14

3.轉速----------------------------------------------------------------------------- 14

4.通氣量-------------------------------------------------------------------------- 15

5.接種量-------------------------------------------------------------------------- 15

三、靈芝固態發酵影響因子-------------------------------------------------- 15

(一)培養基成分------------------------------------------------------------------16

1.營養源-------------------------------------------------------------------------- 17

2.水分----------------------------------------------------------------------------- 18

(二)培養條件---------------------------------------------------------------------18

1.溫度----------------------------------------------------------------------------- 18

2.pH值---------------------------------------------------------------------------- 19

3.通氣與攪拌-------------------------------------------------------------------- 20
4.接菌量-------------------------------------------------------------------------- 21

5.發酵時間----------------------------------------------------------------------- 21

四、自由基與抗氧化------------------------------------------------------------22

(一)自由基對細胞的影響------------------------------------------------------22

(二)自由基與其他分子的反應------------------------------------------------23

(三)抗氧化劑與作用原理------------------------------------------------------24

(四)菇蕈類的抗氧化能力------------------------------------------------------25

五、靈芝對免疫調節的影響---------------------------------------------------27

(一)先天性免疫------------------------------------------------------------------27

(二)後天性免疫反應------------------------------------------------------------28

(三)細胞激素---------------------------------------------------------------------29

(四)靈芝對動物免疫調節的影響---------------------------------------------30

Part I:靈芝固態發酵產物之多醣、β-葡聚醣和抗氧化活性分析-------34

参、靈芝固態發酵對多醣產量及抗氧化活性之探討--------------------35

一、摘要--------------------------------------------------------------------------35

二、前言--------------------------------------------------------------------------35

三、材料與方法-----------------------------------------------------------------37

(一)實驗材料---------------------------------------------------------------------37

(二)實驗儀器---------------------------------------------------------------------38

(三)實驗方法---------------------------------------------------------------------38

1.菌種培養----------------------------------------------------------------------- 38

2.固態發酵培養----------------------------------------------------------------- 38

3.培養基成份對固態發酵的影響-------------------------------------------- 39

4.粗多醣的分析----------------------------------------------------------------- 39

5.β-(1,3)葡聚糖的螢光分析定量-------------------------------------------- 40

6.捕捉1,1-二苯基-2-苦味基團(1,1-diphenyl-2-picryl hydrazyl,DPPH)
能力之測定-----------------------------------------------------------------------41

7.還原力測定-------------------------------------------------------------------- 41

8.螯合亞鐵離子之測定-------------------------------------------------------- 42

9.統計與數據分析-------------------------------------------------------------- 42

四、結果與討論------------------------------------------------------------------42

(一)靈芝固態發酵因子對多醣產量的影響---------------------------------42

(二)靈芝固態發酵產物清除DPPH自由基能力----------------------------45

(三)靈芝固態發酵產物亞鐵離子螯合能力---------------------------------45

(四)靈芝固態發酵產物還原力的檢測---------------------------------------46

五、結論--------------------------------------------------------------------------47

Part II:靈芝固態發酵產物對離乳仔豬生長性狀和細胞激素表現的
影響--------------------------------------------------------------------------------48

肆、靈芝固態發酵產物對離乳仔豬生長性狀及細胞激素表現的影響

一、摘要--------------------------------------------------------------------------49

二、前言--------------------------------------------------------------------------50

三、材料與方法-----------------------------------------------------------------52

(一)實驗材料---------------------------------------------------------------------52

(二)實驗儀器---------------------------------------------------------------------53

(三)實驗方法---------------------------------------------------------------------53

1.動物實驗設計----------------------------------------------------------------- 53

2.腸道黏膜蛋白質、DNA和RNA表現的偵測-----------------------------54

3.反轉錄聚合酶連鎖反應(RT-PCR)----------------------------------------55

四、結果與討論-----------------------------------------------------------------56

(一)餵食不同劑量的靈芝固態發酵產物對離乳仔豬生長性狀的
影響--------------------------------------------------------------------------------56

(二)餵食不同劑量的靈芝固態發酵產物對離乳仔豬腸道黏膜DNA、
RNA和蛋白質測定量的影響-------------------------------------------------58

(三)餵食不同劑量的靈芝固態發酵產物對離乳仔豬淋巴組織細胞
激素表現的影響-----------------------------------------------------------------59

五、結論---------------------------------------------------------------------------63

伍、參考文獻---------------------------------------------------------------------86













表目錄

表1、靈芝有要成分及生理活性--------------------------------------------- 9

表1-1、小麥和黃豆培養基對多醣產量的影響-----------------------------64

表1-2、檢測靈芝固態發酵產物在小麥和黃豆培養基之抗氧化能力- 65

表2-1、細胞激素基因引子序列---------------------------------------------- 73

表2-2、細胞激素PCR片斷大小及循環次數-------------------------------74

表2-3、靈芝固態發酵產物對離乳仔豬生長性能之影響-----------------75

表2-4、靈芝固態發酵產物對離乳仔豬血液生化值之影響(2週齡)----76

表2-5、靈芝固態發酵產物對離乳仔豬血液生化值之影響(4週齡)----77

表2-6、靈芝固態發酵產物對離乳仔豬腸道黏膜DNA測定量
之影響-----------------------------------------------------------------------------78

表2-7、靈芝固態發酵產物對離乳仔豬腸道黏膜RNA測定量
之影響-----------------------------------------------------------------------------79

表2-8、靈芝固態發酵產物對離乳仔豬腸道黏膜蛋白質測定量
之影響-----------------------------------------------------------------------------80















圖目錄

圖1、靈芝的生活史---------------------------------------------------------------4

圖2、自由基對細胞與生物巨分子之傷害-----------------------------------23

圖3、具有免疫調節性質菇蕈類成分對後天性免疫系統之可能作用
標的--------------------------------------------------------------------------------33

圖1-1、以葡萄糖作為多醣檢測標準曲線-----------------------------------66

圖1-2、以Curdlan作為β-1,3-葡聚糖標準曲線-----------------------------67

圖1-3、固態發酵期間多醣含量之變化------------------------------------- 68

圖1-4、固態發酵期間β-葡聚糖含量之變化------------------------------ 69

圖1-5、不同水分比例對多醣產量的影響---------------------------------- 70

圖1-6、不同培養基在第7天和第14天對多醣產量的影響----------- 71

圖1-7、米和薏仁基質靈芝菌絲生長狀態-----------------------------------72

圖2-1、不同淋巴組織之細胞激素表現--------------------------------------81

圖2-2、靈芝固態發酵產物對肺門淋巴結細胞激素mRNA表現量
的影響-----------------------------------------------------------------------------82

圖2-3、靈芝固態發酵產物對脾臟細胞激素mRNA表現量的影響----83

圖2-4、靈芝固態發酵產物對鼠蹊淋巴結細胞激素mRNA表現量
的影響-----------------------------------------------------------------------------84

圖2-5、餵食靈芝固態發酵產物對離乳仔豬肺臟的影響-----------------85
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