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研究生:李德南
研究生(外文):Der-Nan Lee
論文名稱:飼糧中添加甲酸鉻對離乳仔豬生長和免疫反應之影響
論文名稱(外文):Effects of chromium picolinate supplementation on the growth performance and immune responses of weanling pigs
指導教授:陳保基陳保基引用關係沈添富沈添富引用關係
指導教授(外文):Bao-Ji ChenTian-Fuh Shen
學位類別:博士
校院名稱:國立臺灣大學
系所名稱:畜產學研究所
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:1999
畢業學年度:87
語文別:中文
論文頁數:150
中文關鍵詞:生長性能免疫反應離乳仔豬巨噬細胞脂多醣
外文關鍵詞:ChromiumGrowth performanceImmunityWeanling pigsMacrophagesLipopolysaccharide
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鉻可協助哺乳動物之營養分代謝,且飼糧中添加有機鉻可改善受緊迫牛隻之生產和強化免疫反應。本研究之目的,在於建立離乳仔豬獲得最佳生長和免疫反應之鉻量,並探討鉻對離乳仔豬感染期間之生理和免疫反應之影響,續以豬肺臟巨噬細胞為模式,探討鉻對免疫功能提升之機制。
4週齡雜交離乳仔豬餵飼添加(1)0、(2)200、(3)400、和(4)800 ppb的鉻飼糧7週,鉻由甲酸鉻(chromium picolinate, CrPic)提供。結果顯示,飼糧中添加鉻極顯著(P<.01)提高血漿中鉻濃度,添加400 ppb鉻具有提高離乳後4週期間之日增重和飼料利用效率之趨勢。添加鉻雖不影響血漿中之葡萄糖、胰島素、皮質醇濃度及胰島素/葡萄糖之比率,但可改善仔豬離乳後5-7週對假性狂犬病疫苗(Pseudorabies virus, PR)中和抗體力價,及添加鉻具增加第6週免疫球蛋白G(IgG)濃度(P<.15)與總和各週之免疫球蛋白M(IgM)濃度之趨勢(P<.07)。再者於飼糧中添加400 ppb鉻具有增加離乳後4週週邊血液單核細胞分泌白血球中介質-2 (interleukin-2)活性之趨勢(P<0.10),以及降低離乳後6週之血漿三酸甘油酯濃度,但無法改變試驗第6週靜脈注射脂多醣(lipopolysaccharide, LPS) 1μg/kg體重後之血漿中腫瘤壞死因子-α(tumor necrosis factor-α, TNF-α)、皮質醇、葡萄糖及尿素氮濃度。
在試驗第21(d 21)和35日(d 35)仔豬腹腔中注射LPS 200μg/kg體重當作免疫刺激,所有豬隻於注射LPS前(h 0)和後第2 (h 2)、4 (h 4)、8 (h 8)及24小時(h 24)量取直腸溫度及採血,以評估不添加或添加鉻(400 ppb)對緩和免疫緊迫之效果。添加鉻抑制試驗0-21天期間之飼料採食量和顯著降低d 21之體溫0.36℃(P<.05),添加鉻無法改善LPS緊迫下之降低增重影響。LPS注射對血漿代謝物質、荷爾蒙及白血球數目產生劇烈變化,添加鉻促使血漿葡萄糖、三酸甘油酯及尿素氮快速趨向正常,及降低TNF-α於高峰時之濃度和延遲皮質醇之釋出。
添加氯化鉻(chromium chlorite, CrCl3)或CrPic的鉻0至100μg/L與豬隻肺臟巨噬細胞共同培養4小時,不論胰島素存在與否,添加CrCl3的鉻20μg/L,即有最大之葡萄糖攝取率和超氧離子(superoxide anion, )產生。但在胰島素存在下,添加CrPic 的鉻20μg/L即可增加葡萄糖攝取率,且隨CrPic濃度提高而增加產生,而此反應為依賴胰島素途徑。在胰島素存在下,添加CrCl3的鉻10-100μg/L即能顯著提高(P<0.05)巨噬細胞之吞噬大腸桿菌能力,但添加CrPic,不論胰島素存在與否皆不影響。
試驗結果發現,保育期飼糧應推薦添加鉻,添加CrPic的鉻400 ppb,可提高離乳仔豬在正常飼養情況或免疫緊迫下之免疫能力,且根據豬隻肺臟巨噬細胞培養之結果顯示,CrPic可經由胰島素之協助增加巨噬細胞之生理活性。
Chromium (Cr) is an important factor in nutrient metabolism for mammals. Stressed bovine fed with organic forms of Cr has been reported to improve milk production and immune response. The purposes of the study were to evaluate the optimal dietary Cr from picolinate chromium (CrPic) on growth performance and immunity of weanling pigs, the effects of Cr supplementation on the growth and immunity of weanling pigs under endotoxin stress, and the mechanisms of Cr on the cellular activity of pulmonary alveolar macrophages of weanling pigs.
Four-week-old pigs were randomly assigned to (1) 0, (2) 200, (3) 400, and (4) 800 ppb of Cr supplementation group and fed on diets for 7 weeks. Cr supplementation significantly (P<.01) increased plasma Cr, and 400 ppb of Cr supplementation tended to increase daily gain and feed efficiency during the period of 0-4 week postweaning. The levels of plasma glucose, insulin, cortisol, and insulin/glucose ratio were not influenced by various dietary Cr supplementation. Supplementation of Cr increased anti-Pseudorabies virus (PR) neutralizing antibody titers during weeks 5 and 7 (P<.05) and tended to increase IgG level at week 6 (P<.15) and overall means of IgM (P<.07) throughout weeks 2, 4, and 6.
Dietary supplement of 400 ppb of Cr tended to increase IL-2 activity of peripheral blood mononuclear cells on week 4 postweaning and decreased the level of plasma triglyceride on week 6 postweaning, respectively. But plasma TNF-α, cortisol, glucose, and urea nitrogen were not influenced by Cr supplementation after i.v. injection of 1μg/kg body weight of lipopolysaccharide (Escherichia coli; 055: B5, LPS) on week 6 postweaning.
The experiment was performed on two times i.p. injection of 200μg LPS /kg body weight at d 21 and 35, respectively. Plasma samples and rectal temperature were obtained from all piglets before (h 0) and 2 h (h 2), 4 h (h 4), 8 h (h 8), or 24 h (h 24) after LPS injection. The effect of Cr supplementation (400 ppb) on weanling pigs after LPS challenge was also determined. The supplementation of Cr before LPS injection (d 0-21) decreased the feed intake (0.42 vs. 0.34 kg/day) and rectal temperature at d 21 was decreased significantly (P<0.05) about 0.36℃. Supplementation with Cr had no effect on alleviating the growth performance after LPS challenges. The plasma metabolites, hormones, and immune response changed dramatically after the initiation of LPS injection. Supplementation with Cr facilitated the recovery of plasma glucose, triglyceride, and urea nitrogen rapidly. Plasma TNF-α level was reduced and cortisol release was delayed with Cr supplementation after LPS challenge at d 35.
The pulmonary alveolar macrophages were incubated with 0-100μg/L of two Cr salts from chromium chloride (CrCl3) and CrPic for 4 hrs at 37℃. The glucose uptake and superoxide anion () production were increased in the cells incubated with 20μg/L of Cr from CrCl3. The glucose uptake induced by CrPic might be an insulin-dependent manner. Supplementation of Cr from CrPic at a concentration of 100μg/L increased production significantly (P<0.05) regardless of the presence of insulin. In the presence of insulin, the phagocytosis of E. coli by macrophages was enhanced significantly (P<0.05) in medium supplemented with 10-100μg/L of Cr from CrCl3 and CrPic supplementation did not affect the phagocytosis of macrophage regardless of the presence of insulin.
The results suggest that supplemental 400 ppb of Cr from CrPic was beneficial during normal feeding condition and under LPS stress in weanling pigs. Supplementation of CrPic under the cooperated action of insulin enhances the cellular activity of macrophages.
I、摘要------------------------------------------------ 1
II、前言----------------------------------------------- 3
III、文獻檢討------------------------------------------ 5
一、鉻對家畜生產之影響-------------------------------- 5
(一)、鉻研究之歷史------------------------------------- 5
(二)、鉻之代謝和功能特性------------------------------- 7
1.鉻之消化與吸收途徑-----------------------------------7
2.鉻對血糖、脂肪及蛋白質代謝之調節--------------------- 9
3.鉻對家畜生產和屠體性狀之影響-------------------------10
4.鉻之抗緊迫功能---------------------------------------12
二、影響仔豬免疫能力之因素----------------------------15
(一)、豬隻免疫系統之發育--------------------------15
(二)、仔豬離乳之緊迫------------------------------17
(三)、飼糧抗原之影響------------------------------18
(四)、脂多醣毒素之影響----------------------------19
1.LPS對家畜荷爾蒙和細胞激素持恆之影響-----------------20
2.LPS對家畜生長、生理及營養分代謝之影響---------------23
3.LPS對免疫反應之影響---------------------------------25
三、影響巨噬細胞葡萄糖代謝之因素---------------------26
(一)、細菌性刺激原之作用-------------------------28
(二)、胰島素之影響-------------------------------29
(三)、鉻離子之作用-------------------------------31
IV、試驗部份-------------------------------------------33
一、 啶甲酸鉻添加量對離乳仔豬生長和免疫反應之
影響---------------------------------------------------33
前言---------------------------------------------------34
材料與方法---------------------------------------------35
結果與討論---------------------------------------------39
二、飼糧中添加啶甲酸鉻對豬隻週邊血液單核細胞
分泌白血球中介質-2和注射脂多醣之生理與免疫
反應---------------------------------------------------51
前言---------------------------------------------------52
材料與方法---------------------------------------------53
結果與討論---------------------------------------------58
三、飼糧中添加鉻對免疫緊迫下離乳仔豬生長和免疫
反應之影響---------------------------------------65
前言---------------------------------------------------66
材料與方法---------------------------------------------67
結果與討論---------------------------------------------72
四、氯化鉻和甲酸鉻對豬隻肺臟巨噬細胞生理活性
之影響---------------------------------------------91
前言---------------------------------------------------91
材料與方法---------------------------------------------93
結果與討論---------------------------------------------96
V、結論--------------------------------------------- 108
VI、參考文獻----------------------------------------- 109
VII、英文摘要------------------------------------------138
VIII、附錄-------------------------------------------- 141
一、血漿IgG濃度之測定------------------------------ 141
二、週邊血液單核細胞(PBMC)增生反應之測定------------ 143
三、週邊血液單核細胞分泌IL-2活性之測定-------------- 145
四、血漿中胰島素濃度之測定-------------------------- 146
五、血液淋巴球表面抗原之測定------------------------ 147
IX、小傳---------------------------------------------- 150
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