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研究生:林義福
研究生(外文):Yih-Fwu Lin
論文名稱:飼糧添加維生素E對台灣土雞繁殖性能、免疫反應及抗氧化之影響
論文名稱(外文):Effects of dietary vitamin E supplementation on reproduction, immune response and antioxidant status of Taiwan native chickens
指導教授:張素瓊張素瓊引用關係
指導教授(外文):Sue-Joan Chang
學位類別:博士
校院名稱:國立成功大學
系所名稱:生命科學系碩博士班
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:英文
論文頁數:132
中文關鍵詞:繁殖免疫抗氧化維生素E
外文關鍵詞:vitamin Echickensimmuneantioxidantreproduction
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  • 下載下載:119
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摘 要

  維生素E最先是被發現為大鼠正常繁殖所需之營養成份,隨後亦發現為維持細胞膜完整性所需,並且與免疫功能有關,由於有關維生素E對雞之影響的資料並不多,本研究以台灣種土雞為試驗動物,分別探討飼糧添加維生素E對(1)母雞繁殖(2)公雞繁殖 (3)免疫反應及(4)抗氧化之影響。

  試驗1探討不同維生素E添加量對母雞繁殖之影響。以一日齡母雛雞300隻餵飼玉米-大豆粕為主飼糧至17週齡,17週齡後,逢機區分為五處理組,餵飼以玉米-大豆粕為主飼糧,分別添加0、40、80、120及160 mg/kg維生素E (DL-a-tocopheryl acetate),飼養至46週齡。另一批15隻公雞則餵飼一般實用飼糧,做採精及人工授精用。結果顯示,添加120 mg/kg維生素E顯著降低初產蛋重,但對初產日齡、初產體重或至46週齡止之死亡率,各組間並無顯著差異。添加維生素E提高母雞於產蛋高峰期之增重,添加80 mg/kg維生素E顯著改進飼料換蛋率及產蛋量,並於產蛋高峰後顯著增加產蛋率,但添加量高於80 mg/kg時則效益降低。於17至46週齡間,蛋品質(蛋比重及蛋殼強度)隨週齡增加而降低。在試驗期間,週齡與受精率、孵化率則無相關,顯示種母雞之蛋品質比繁殖性能易受週齡之影響。與對照組比較,添加80 mg/kg維生素E組之受精率與孵化率分別增加7.7及13.4%,受精蛋之孵化率則無差異。本結果顯示,產蛋期添加維生素E可提高種母雞繁殖性能,其中添加80 mg/kg維生素E組有最佳產蛋率、產蛋量、飼料換蛋率、孵化率及受精率。

  試驗2探討不同維生素E添加量對公雞繁殖之影響。一日齡公雛雞90隻餵飼玉米-大豆粕為主飼糧至23週齡,23週齡週齡後,逢機區分為五處理組,餵飼以玉米-大豆粕為主飼糧分別添加0、20、40、80及160 mg/kg維生素E (DL-a-tocopheryl acetate),飼養至52週齡。另一批225隻同齡母雞則餵飼一般實用飼糧,做公雞繁殖能力測定用。母雞於31至43週齡間以未稀釋公雞精液(0.04 mL/隻)作人工受精,於49週齡以稀釋5倍相同量(0.04 mL/隻)之公雞精液作人工受精。結果顯示,添加維生素E於試驗期間並不會影響公雞之有效授精持續力及授精率,然而,添加160 mg/kg維生素E組顯著增加最大授精持續力,(49週齡,5倍稀釋精液) 。添加40至160 mg/kg維生素E,公雞於39週齡後有顯著較高之精蟲存活率及活力,其中添加80 mg/kg維生素E組,於39週齡有顯著較高之精蟲濃度。公雞餵飼添加40 mg/kg以上之維生素E,於試驗期間有較高之增重。血漿膽固醇及睪固酮不受添加維生素E之影響,但添加160 mg/kg維生素E組之血漿促黃體生成激素(LH)顯著較低,本試驗結果顯示,公雞長期(52週)餵飼不添加維生素E飼糧,會使精液品質降低,但不損害授精率,此一結果是否因授精量彌補了精液品質,有待進一步探討。

  試驗3測定維生素E對公雞及母雞之免疫反應。試驗設計如試驗1及試驗2所示,疫苗接種依一般商業雞場之種類、方法及日程進行。測定項目包括:抗綿羊紅血球(SRBC)、新城雞病病毒(NDV)、傳染性華氏囊病病毒(IBDV)、傳染性支氣管病病毒(IBV) 等抗體力價及對植物血凝素(PHA-P)之皮膚腫脹反應。結果顯示,飼糧添加維生素E並不影響產蛋母雞免疫反應,對公雞而言,飼糧添加20及40 mg/kg維生素E可提高抗SRBC抗體力價,添加20 mg/kg維生素E有最高抗IBDV力價最高,然而,糧添加80及160 mg/kg維生素E有較低之抗SRBC及IBDV力價。本試驗結果顯示,適量添加維生素E對某些抗原如SRBC可促進其免疫反應,然而,對某些抗原如IBDV,當添加量超出80 mg/kg時可能有抑制效果,本試驗因此建議,玉米-大豆粕飼糧添加20 mg/kg維生素E來提高公雞免疫反應是合宜的。

  試驗4測定母雞及孵化雛雞之抗氧化能力。試驗設計如試驗1所示,母雞於35週齡測定添加維生素E對雛雞抗氧化能力之影響,逢機抽取12隻孵化雛雞,犧牲後取腦及肝臟做抗氧化分析,包括脂質過氧化指標-malondialdehyde (MDA)量、氧化壓力指標-反應性氧化物(ROS)量、過氧化氫酶(catalase)及過氧化物歧化酶(superoxide dismutase)活性等。結果顯示,母雞血漿維生素E濃度隨飼糧維生素E添加量增加而直線增加(P < 0.001; r = 0.997),但蛋黃維生素E濃度,於添加120 mg/kg維生素E時即達最高。維生素E添加量增加,母雞血漿及雛雞腦MDA之量顯著降低(P < 0.05)。雛雞腦及雛雞肝臟ROS之量亦有相同之結果。於抗氧化酵素方面顯示,母雞給予120及160 mg/kg維生素E,會分別顯著增加(P < 0.05)雛雞肝臟過氧化氫酶及雛雞腦過氧化物歧化酶之活性。本試驗結果顯示,添加之維生素E易轉移到母雞血漿及蛋黃中,此外,母雞添加高量維生素E(120與160 mg/kg),有益於提高孵化雛雞之抗氧化力,降低氧化壓力。

  總合以上結果顯示,維生素E對母雞繁殖之影響大於對公雞之影響。母雞添加維生素E可降低母雞與孵化雛雞之氧化壓力,並提高雛雞抗氧化力。適度添加維生素E可促進公雞抗綿羊紅血球之免疫反應,然而添加高量維生素E對特定抗原如IBDV會降低其反應。
 Vitamin E was originally described as a dietary constituent required for normal reproduction in rats. Subsequently, it was recognized as an essential vitamin for maintaining the integrity of biological cell membranes. Vitamin E was also one of the nutrients that have been reported playing a role in immune function. Due to the rare data in its effects on chicken, effects of dietary vitamin E on 1) female reproduction, 2) male reproduction, 3) immune response and 4) antioxidant status were studied in Taiwan native breeder chickens.
 
 Experiment I — female reproduction. This study was conducted to determine adding graded levels of vitamin E after the onset of laying on the reproductive outcome of breeder pullets.

 Day-old female chicks were fed on corn-soybean growing diets without supplemental vitamin E from hatch to 17 weeks of age. After 17 weeks the pullets (n = 300) were randomly assigned to 5 dietary treatments and fed on corn-soybean laying diets supplemented with 0, 40, 80, 120 and 160 mg/kg of vitamin E (dl-α-tocopherol acetate), respectively, until 46 weeks of age. A flock of 15 cockerels were fed with practical diets throughout the experiment for semen collection and artificial insemination (AI).

 The results for female reproduction indicated that addition of 120 mg/kg of vitamin E lowered the first egg weight (P < 0.05); however, there was no significant difference in the age or body weight of pullets at first egg or mortality rate to 46 weeks of age among the treatments. Supplemental vitamin E improved body weigh gain of laying hens during peak-laying period. Feed efficiency and egg mass were improved (P < 0.05) in pullets fed 80 mg/kg of supplemental vitamin E. A significant increase in egg production was observed after peak egg production in pullets given 80 mg/kg of supplemental vitamin E. However, this favorable effect decreased as supplemental vitamin E exceeded 80 mg/kg. Egg quality decreased with age. There was no correlation between age and fertility or hatchability during the experimental period, suggesting that egg quality is more age-sensitive than reproductive performance for breeder pullets. Compared with the control, fertility and hatchability of all eggs set for the treatment with 80 mg/kg supplemental vitamin E increased by 7.7 and 13.4%, respectively. There was no difference in the hatchability of fertile eggs. These results suggest that using supplemental vitamin E during the laying period can improve the reproductive performance of breeder pullets. The addition of 80 mg/kg of vitamin E obtained the best performance in egg production, egg mass, feed efficiency, hatchability and fertility.

 Experiment II —male reproduction. This study was conducted to investigate effects of supplemental vitamin E on reproduction of cockerels.

 Day-old male chicks were fed with corn-soybean rearing diets without supplemental vitamin E for 23 weeks. The cockerels (n = 90) were then randomly assigned to five groups and fed with the corn-soybean basal diets supplemented with either 0, 20, 40, 80 or 160 mg/kg of vitamin E (dl-a-tocopherol acetate) for 29 weeks. A flock of 225 pullets were fed with practical diets throughout the experiment. They were artificially inseminated with one dose of intact and five-fold diluted pooled semen (0.04 mL/bird) at 31 to 43 weeks and at 49 weeks of age, respectively.

 The results for male reproduction indicated that supplemental vitamin E did not affect cockerels’ effective duration of fertility and percentage of fertility throughout the experiment. However, supplementing 160 mg/kg vitamin E significantly increased the maximum duration of fertility at 49 weeks of age, when pullets were inseminated with diluted semen. Cockerels fed diets supplemented with 40 to 160 mg/kg vitamin E had significantly higher sperm viability and motility after 39 weeks of age and those fed 80 mg/kg vitamin E had significantly higher sperm concentration at 39 weeks of age. Cockerels fed on diets supplemented with more than 40 mg/kg vitamin E had higher body weight gain in the experimental period. This results suggest that long-term (52 weeks) without supplemental vitamin E was associated with lower semen quality in the cockerels. Although it did not cause vitamin E deficiency syndrome. We could not demonstrate that lack of supplemental vitamin E caused impairment of male fertility, in terms of the proportion of fertile eggs laid by inseminated hens, since the insemination dose may have compensated for low sperm quality. We did find that the maximim duration of fertility might be improved by supplementing 160 mg/kg vitamin E at 49 weeks of age.

 Experiment III — immune response. This study was conducted to investigate effects of supplemental vitamin E on immune status of female and male birds.

 Supplemental vitamin E did not affect immune response of laying hens. For cockerels, dietary addition of 20 and 40 mg/kg vitamin E had a positive effect on immune response to sheep red blood cell (SRBC) and supplementation of 20 mg/kg vitamin E had the highest antibody titer to infectious bursal disease virus (IBDV). However, supplementation of 80 to 160 mg/kg vitamin E had lower immune response to SRBC and IBDV. These results suggest that moderate supplementation of vitamin E had shown to enhance immune responses to SRBC, whereas for selective antigens such as IBDV depression may occur when supplementation level is over 80 mg/kg. We suggest that supplementation of 20 mg/kg vitamin E to corn-soybean diet was optimal for enhancing immune response of cockerels.

 Experiment IV — antioxidant status. This study was conducted to investigate effects of supplemental vitamin E on antioxidant status of female birds and hatching chicks. Hatching chicks form each treatment were sacrificed for evaluating effects of maternal vitamin E supplementation on antioxidant status of chicks.

 Plasma vitamin E concentration of pullets increased linearly (P < 0.001; r = 0.997) as the increase of supplemental vitamin E, but egg yolk concentration reached a plateau at 120 mg/kg supplemental vitamin E. The malondialdehyde (MDA) levels, an indicator of lipid peroxidation, of hen plasma and chick brain significantly decreased as the increase of supplemental vitamin E (P < 0.05). Same results were found in the reactive oxygen species (ROS) levels, an indicator of oxidative stress, of chick brain and liver. For antioxidant enzymes, pullets given 120 mg/kg and 160 mg/kg of supplemental vitamin E significantly increased (P < 0.05) activities of catalase (CAT) of chick liver and superoxide dismutase (SOD) of chick brain, respectively. These results indicated that supplemental vitamin E could easily transfer to plasma and egg yolk of pullets. In addition, maternal supplementation of high levels of vitamin E (120 to 160 mg/kg) has beneficial effects on enhancing antioxidant capability and depressing oxidative stress of hatching chicks.

 In conclusion, these data suggest that supplemental vitamin E improve reproductive performance of female chickens better than male chickens. Supplemental vitamin E exhibit beneficial effects on enhancing antioxidant capability and depressing oxidative stress of hatching chicks. Moderate supplementation of vitamin E may enhance immune responses to SRBC, whereas for selective antigens, such as IBDV, depression may occur by high supplemental vitamin E.
TABLE OF CONTENTS

ENGLISH ABSTRACT I
CHINESE ABSTRACT V
ACKNOWLEDGEMENTS IX
TABLE OF CONTENTS X
LIST OF TABLES XIII
LIST OF FIGURES XVI
ABBREVIATIONS XVII

INTRODUCTION 1
LITERATURE REVIEW
Vitamin E Discovery and History 3
Vitamin E Structure 3
Vitamin E Biological Activity 4
Feed Sources 5
Stability of Vitamin E 6
Absorption and Uptake 7
Delivery 9
Metabolism 11
Storage 11
Excretion 12
Vitamin E and Reproduction 12
Antioxidant function of vitamin E 14
Pro-Oxidant Activity of Vitamin E 14
Vitamin E and Immunity 15
Factors Affecting Vitamin E Requirements 16
Vitamin E Deficiency 16
Effects of vitamin E excess 17

EXPERIMENTS: Part I to IV.
I. Effects of supplemental vitamin E during the laying period on the reproductive performance of Taiwan native breeder pullets
1. Abstract 18
2. Introduction 20
3. Materials and Methods 21
4. Results 25
5. Discussion 28
II. Effects of supplemental vitamin E during the mature period on the reproductive performance of Taiwan native breeder cockerels
1. Abstract 42
2. Introduction 44
3. Materials and Methods 45
4. Results 49
5. Discussion 51
III. Effects of supplemental vitamin E on immune response of Taiwan native breeder chickens
1. Abstract 65
2. Introduction 67
3. Materials and Methods 68
4. Results 72
5. Discussion 75
IV. Effects of maternal vitamin E supplementation on antioxidant status of chicks
1. Abstract 87
2. Introduction 89
3. Materials and Methods 90
4. Results 96
5. Discussion 97

SUMMARY AND CONCLUSIONS 105
LITERATURE CITED 107
PUBLICATION LIST 129
CURRICULUM VITAE 131
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