臺灣博碩士論文加值系統

本試驗之目的旨在探討於牛隻分娩前2-3週注射β-carotene、Vitamin E或硒，是否對分娩前後牛隻血液中β-carotene、α-tocopherol和硒濃度之維持、泌乳初期臨床性乳房炎的發生率、體細胞數及嗜中性球之噬菌能力有顯著影響，以期能利用此等營養素之施用，增強牛隻乳房之免疫功能，有效降低牛乳體細胞數、臨床性乳房炎的發生率，以減少酪農業者之經濟損失，而增進其收益。28頭荷蘭母牛於分娩前2-3週分成5組，4組投藥組各分別予以肌肉注射(1)β-carotene ( 1.5 mg/ 100 Kg of body weight) + Vitamin E (3,000 IU/ cow) + Sodium Selenite (50 mg/ cow, 含22.4 mg Se) （BCESE），內含佐劑Tween 80 和saseme oil量如TWSO、(2) Vitamin E (3,000 IU/ cow) + Sodium Selenite (50 mg/ cow)（ESE），內含佐劑Tween 80量如TW、(3) Tween 80 (4.4 g/ cow)（TW）、(4) Tween 80 (4.4 g/ cow) + sesame oil ( 3 mL / 100 Kg of body weight)（TWSO），或(5)控制組（CONTR）不予任何處理。於注射前，注射後1、4、7天和分娩後24小時內及分娩後第1、4、7天進行血液、乳液之β-carotene、α-tocopherol和硒濃度測定，分娩後1個月內進行產乳性狀、嗜中性球噬菌能力、臨床性乳房炎發生率之檢查。試驗結果顯示牛隻注射前血漿中β-carotene濃度皆在5μg/mL以上，注射後所測量到之平均血漿β-carotene濃度分別為：BCESE為5.59μg/mL，ESE組為5.09μg/mL，TW為4.77μg/mL，TWSO為5.70μg/mL，CONTR為5.74μg/mL。注射前後各處理組間平均血漿中β-carotene濃度並無顯著差異。分娩後各處理組血漿之平均β-carotene濃度分別為：BCESE為3.07μg/mL，ESE為3.24μg/mL，TW為2.90μg/mL，TWSO為3.30μg/m，CONTR為2.66μg/mL，分娩前後所有牛隻血漿中β-carotene濃度皆高於正常值（1.5μg/mL），表示所有受試牛於試驗期間β-carotene之攝食量均已充足。注射處理後BCESE和ESE之平均血漿中α-tocopherol濃度均顯著高於未注射Vitamin E之TW、TWSO和CONTR者（P＜0.05）。分娩時未注射Vitamin E之TW、TWSO、CONTR之牛隻血漿中α-tocopherol濃度會迅速下降，顯示注射Vitamin E者可維持其血漿中α-tocopherol的濃度使之不至急遽下降，且其乳中α-tocopherol的濃度顯著高於未注射Vitamin E者（P＜0.05）。試驗處理對牛隻血清及乳中硒之濃度、全血中Glutathione peroxidase活性並無顯著影響。BCESE和ESE牛隻分娩後第二週之生乳體細胞數，較低於TW、TWSO與CONTR者。但各試驗處理對牛隻分娩後嗜中性白血球吞噬大腸桿菌和金黃色葡萄球菌的能力、乳房炎的發生率及乳量並未有顯著差異存在。
本試驗所有受試牛之β-carotene和硒的營養狀態均良好，此或許是牛隻於分娩前注射β-carotene和硒未能產生如預期效果之原因。惟分娩前注射Vitamin E可減緩牛隻分娩直後血漿及乳液中α-tocopherol的濃度劇烈下降的程度。

The purposes of this study was to verify whether there would be effects of intramuscular injection of β-carotene, vitamin E, and selenium during 2-3 weeks preparturition on sustaining high blood levels of these nutrients, decreasing incidence of clinical mastitis and somatic cell counts (SCC), and increasing neutrophil phagocytosis during early lactation in Holstein cow. It was expected that the application of these nutrients would enhance the immune function in mammary glands of dairy cows and consequently declined the economic loss attributable to the udder problems.
Twenty eight cows were randomly allotted into one of the five treatments, among them, four medicated groups had been respectively given intramuscular injection of : 1) 1.5 mg β-carotene per 100 Kg body weight plus 3,000 IU vitamin E and 50 mg sodium selenite (induding 22.4 mg Se) per head (BCESE), including the adjuvants, Tween80 and sesame oil (dosage as per TWSO), 2) 3,000 IU vitamin E and 50 mg sodium selenite per head (ESE), including the adjuvant, Tween80 (dosage as per TW), 3) 3 mL sesame oil (solvent forβ-carotene ) per 100 Kg body weight plus 4.4 g Tween 80 per head (TWSO), 4) 4.4 g Tween 80 per head (solvent for vitamin E; TW), or no injection (Control) during 2-3 weeks prior to calving. From the concentration test of β-carotene, α-tocopherol, selenium and glutathione peroxidase in blood or milk conducted on the first, fourth and seventh pre- and post-injection days, within postparturition 24 hours and the first, fourth and seventh days postparturition, in addition to the examination proceeded within one month postparturition on milk yield, neutrophilic bacteriophagology and incidence of clinical mastitis. At preinjection, the blood plasma β-carotene concentration in each of the 5 treatments was above 5 μg/mL. During postinjection, the blood plasma β-carotene concentration in BCESE, ESE, TW, TWSO, and CONTR were 5.59 μg/mL, 5.09μg/mL, 4.77 μg/mL, 5.70 μg/mL, and 5.74 μg/mL, respectively. There was no significant difference in the blood plasma β-carotene concentration between preinjection and postinjection in all the treatments. During postparturition, the blood plasma β-carotene concentration in BCESE, ESE, TW, TWSO, and CONTR were 3.07 μg/mL, 3.24 μg/mL, 2.90 μg/mL, 3.30 μg/mL, and 2.66 μg/mL, respectively. The blood plasma β-carotene concentration of the cows in all treatments was above normal level, 1.5 μg/mL, during periparturition indicating that these cows were sufficient in β-carotene nutritional status. The average of BCESE and ESE in postinjection blood plasma α-tocopherol concentration was greater (P<0.05) than those without injection of vitamin E. Blood plasma α-tocopherol concentration declines abruptly during parturition. Cows injected with vitamin E could prevent from rapid decrease of the nutrient concentration in blood and increase the nutrient concentration in milk (P<0.05). The treatments exerted no significant effect on selenium concentration in blood serum and milk, and glutathione peroxidase activity in whole blood. The SCC at week 2 postparturition in BCESE and ESE were less than those in TW, TWSO, and CONTR. However, there were no significant differences observed in neutrophil phagocytosis to Escherichia Coli and Staphylococcus aureus, incidence of mastitis and milk yield of the cows.
In conclusion, the efficacy of injecting β-carotene and selenium to cows during preparturition was not observed as expecting and the rationale for this might be attributable to the sufficient nutritional status of these nutrients in the cows under examined. Nevertheless, injection of vitamin E to cows during preparturition was effective in decreasing the severity of abruptly drop of α-tocopherol concentrations in blood and in milk right after calving.

目錄
壹 中文摘要………………………………….………………………….1
貳 文獻檢討………………………………………….……………….... 3
一 乳牛之乳房炎………………………………………...………….3
（一）乳房炎之定義…………………………………………….3
（二）乳房炎之分類與症狀……………………………………..3
（三）乳房炎之發生原因……………………………..………..5
二 體細胞數與牛隻乳腺健康之關係...…………………….......…..7
（一） 體細胞之簡介…………………………………...……7
（二） 體細胞數與乳量之關係…………………….….…….9
（三） 影響體細胞數的因素……………………….….…….10
三 嗜中性球及其免疫作用.…………………………..……..…….12
（一） 嗜中性球之簡介…………………………………….12
（二） 血液與乳液之嗜中性球型態學之比較…………….13
（三） 嗜中性球的吞噬作用……………………………….14
（四） 嗜中性球對乳腺之免疫作用…………………...…..14
四 β-carotene及其生物功能…………………………………..….15
（一） β-carotene的來源…………………………...……..15
（二） β-carotene的代謝……………………………….…16
（三） β-carotene的抗氧化功能………………………….17
（四） β-carotene的免疫功能………………………...…..17
（五） β-carotene對乳腺健康及繁殖性能的影響…….....18
五 Vitamin E及其生物功能…………………………………...…...20
（一） Vitamin E的吸收與移轉…………………………....20
（二） Vitamin E的生理功能………………………………21
（三） Vitamin E對乳牛乳腺健康的影響…………………21
六 硒及其生物功能…………………………………………….......23
（一） 硒之簡介…………………………………………….23
（二） 硒在體內之分佈………………………………...…..24
（三） 硒的代謝…………………………………...………..24
（四） 硒的生物功能……………………………………….26
（五） 硒對牛隻乳腺健康之影響………………...………..27
參 前言……………………………………………………………...….29
肆 試驗材料與方法……………………………………………………31
一 試驗動物……………………………………………………..…31
二 實驗處理………………………………………………………..31
三 樣品採集及製備………………………………………………..32
四 Vitamin E和β-carotene含量之分析…………………….……33
（一）β-carotene…………………….………………………..33
（二）Vitamin E………………………..………………………34
五 硒含量的測定…………………………………………………..35
六 全血中Glutathione peroxidase活性之測定…………………...35
七 嗜中性球噬菌能力之測定…………………………………..…36
（一） 血中嗜中性白血球之分離………………………….36
（二） 細菌之製備………………………………………….37
1.菌種保存…………………………………….…….37
2. 細菌處理………………………..……….………...37
（三） 噬菌能力之測定…………………………..………....38
八 乳量…………………………………………………….…..…….38
九 生乳體細胞數之測定…………………………………….…….38
十 臨床性乳房炎………………………………………………..…39
十一 統計分析………………………………………………….…...39
伍 結果與討論………………………………………………………..40
一 各試驗階段乳牛血漿及乳汁中β-carotene
濃度之變化…………………………………………………...….40
二 各試驗階段乳牛血漿及乳汁中α-tocopherol
濃度之變化…………………………………………………...….44
三 各試驗階段乳牛血漿及乳汁中硒濃度之變化………..……..…46
四 各試驗階段乳牛全血中Glutathione peroxides
(GSH-Px)濃度之變化…………………………………………..53
五 注射β-carotene、Vitamin E和硒對牛隻乳液中
體細胞數之影響………………………………………………..56
六 注射β-carotene、Vitamin E和硒對牛隻臨床性
乳房炎發生率之影響……………………………………….….58
七 注射β-carotene、Vitamin E和硒對牛隻產乳性狀之影響….58八 注射β-carotene、Vitamin E和硒對牛隻嗜中性
球噬菌能力之影響………………………………………..…….59
陸 結論………………………………………………………………..66
柒 參考文獻…………………………………………………………..67
捌 英文摘要…………………………………………………………..78
玖 作者小傳…………………………………………………………..81
表次
表1. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血漿及乳中β-carotene (μg/mL)濃度平均值之影響…….41
表2. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血漿中β-carotene (μg/mL)濃度之影響……………...…....42
表3. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其乳中β-carotene (μg/mL)濃度之影響………….………....43
表4. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血漿及乳中α-tocopherol (μg/mL)濃度平均值之影響…..45
表5. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血漿中α-tocopherol (μg/mL)濃度之影響……………...…47表6. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其乳中α-tocopherol (μg/mL)濃度之影響………………...…48
表7. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血清及乳中硒(ng/mL)濃度平均值之影響………...….….49
表8 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其血清硒(ηg/mL)濃度之影響………………………..…….51
表9荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其乳中硒(ηg/mL)濃度之影響……………………………...52
表10荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其全血中GSH-Px (Ug/mL)活性平均值之影響………….....54
表11 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其全血中GSH-Px(Ug/mL)活性之影響…………..……...….55
表12. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其體細胞數、臨床性乳房炎發生率及產乳量之影響………57
表13. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其分娩後每個嗜中性球吞噬金黃色葡萄球菌菌數之影響..64
表14. 荷蘭母牛於分娩前行肌肉注射β-carotene、Vitamin E和硒
對其分娩後每個嗜中性球吞噬大腸桿菌菌數之影響………..65
圖次
圖1、 分娩前後牛隻血清中Vitamin A和β-carotene濃度之變化…...19
圖2、臨近分娩前後時期血漿中Vitamin E濃度之變化……………...22
圖3、荷蘭母牛硒的採食量與其血漿及乳液中硒濃度間之關係.……25
圖4、荷蘭母牛血漿硒濃度與總乳體細胞數間之關係…….………...28
圖5、由罹患臨床型乳房之荷蘭母牛乳液中分離出
之(a)金黃色葡萄球菌(b)大腸桿菌…………………………...61
圖7、光學顯微鏡下嗜中性球吞噬金黃色葡萄球菌之情形………...62
圖8、 光學顯微鏡下嗜中性球吞噬大腸桿菌之情形…………….…....63

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