蛋白质与氨基酸营养对早期断奶犊牛免疫相关指标的影响
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摘要
随着我国奶业的发展,利用植物蛋白部分代替乳蛋白集约化饲养犊牛已经获得初步成功,但植物蛋白对犊牛免疫机能的影响效果及如何提高植物蛋白的氨基酸平衡和利用效果仍然有待研究,因此,我们的目标就是通过研究蛋白质营养对犊牛免疫机能的影响,通过改善限制性氨基酸平衡以提高犊牛对植物蛋白的利用效果。研究结果摘要如下:
     1蛋白质水平对断奶犊牛免疫及相关指标的影响
     本试验选择初生荷斯坦公犊15头,随机分为三组,分别饲喂蛋白质水平为18%、22%和26%的代乳品,研究蛋白质水平对断奶犊牛免疫及相关指标的影响。结果表明:蛋白质水平为22%时,犊牛血清IgG含量最高(P>0.05),血清AKP达到峰值,体重和脾脏指数最大;蛋白质水平高于22%时,犊牛炎性细胞因子(IL-1)显著升高(P<0.05),犊牛小肠肥大细胞数量显著增加(P<0.05);犊牛血清GH(P>0.05)、IGF-1(P<0.05)、TAOC(P<0.05)等指标有随蛋白质水平的提高而升高的趋势,但过高的蛋白质水平降低了犊牛NO水平;22%的蛋白质水平有利于促进犊牛蛋白质合成代谢与生长发育,增强机体免疫能力,蛋白水平过高或过低均不利于犊牛生长和机体免疫机能。
     2蛋白质组成对断奶过渡期犊牛免疫相关指标的影响
     本试验选择初生荷斯坦公犊12头,随机分为三组,分别饲喂植物蛋白占总蛋白20%、50%和80%的代乳品,研究蛋白质组成对断奶过渡期(6~11日龄)血液相关指标的影响。结果表明:植物蛋白占总蛋白的50%时,断奶过渡期犊牛血清总蛋白、GH、IgG最高(P>0.05),尿素氮最低(P>0.05),血清TAOC、NO、AKP等指标水平最高(P<0.05);植物蛋白超过总蛋白的50%时,犊牛腹泻率和腹泻频率显著升高(P<0.05),犊牛腹泻情况在断奶过渡期前3天比较严重。
     3蛋白质组成对断奶后犊牛免疫相关指标的影响
     本试验选择初生荷斯坦公犊12头,随机分为三组,分别饲喂植物蛋白占总蛋白20%、50%和80%的代乳品,研究蛋白质组成对断奶后(2~8周龄)犊牛血液相关指标的影响。结果表明:植物蛋白占总蛋白的50%时,断奶后犊牛血清IgG、IL-1和TNF-α水平最高(P>0.05),AKP水平最高;2~4周龄,犊牛IgG、IL-1和TNF-α水平呈下降趋势,4~8周龄则呈上升趋势;植物蛋白含量的提高,降低了小肠粘膜粘膜SIgA水平和肥大细胞数量(P<0.05),提高了杯状细胞数量,降低了犊牛血清皮质醇水平,而对溶菌酶、GH和IGF-1没有显著影响;植物蛋白含量为80%时,提高了犊牛腹泻率(P<0.05)和腹泻频率(P>0.05);犊牛在4周龄以后对50%及以下的植物蛋白表现出较好的适应性。
     4主要限制性氨基酸对犊牛免疫及生产性能的影响
     本试验选择20头荷斯坦公犊牛随机分为四组,分别饲喂四种采用氨基酸部分扣除法配制的不同氨基酸比例模式的代乳品,研究赖氨酸、蛋氨酸和苏氨酸缺乏对犊牛生产性能和免疫相关指标的影响。结果表明:限制性氨基酸缺乏降低了犊牛采食量、体重、日增重,降低了犊牛氮的采食、氮沉积、氮表观消化率和表观生物学价值等,其影响程度以赖氨酸最大,其次是蛋氨酸、苏氨酸;对于犊牛非特异性免疫反应,赖氨酸缺乏时其反应提高,蛋氨酸缺乏时在犊牛28日龄时降低而56日龄时升高,苏氨酸缺乏则影响较小;赖氨酸和蛋氨酸缺乏,使得28日龄犊牛免疫系统由Th2向Th1偏移,犊牛细胞免疫水平提高,体液免疫水平降低,而56日龄犊牛免疫系统则发生了相反的变化,由Th1向Th2偏移,犊牛体液免疫水平提高,与犊牛血清IgG浓度的变化一致,显著提高56日龄犊牛血清IgG水平;苏氨酸缺乏对28日龄和56日龄犊牛免疫系统则均是由Th1向Th2偏移,犊牛体液免疫水平提高;苏氨酸缺乏组犊牛胸腺、脾脏、肝脏指数均有降低趋势;赖氨酸和蛋氨酸缺乏犊牛的腹泻率、腹泻频率、粪便指数均明显提高。
     5免疫应激对犊牛限制性氨基酸需求模式的影响
     本试验选择40头荷斯坦公犊牛,随机分为两个组,分别为正常组、应激组,每组内设正对照组和三个扣除氨基酸的试验组,采用氨基酸部分扣除法比较LPS诱导的免疫应激对断奶犊牛的Lys、Met和Thr三种主要限制性氨基酸平衡模式的影响。结果表明:免疫应激时犊牛体温、血清IL-1和TNF-α、血清NO均显著升高,血清补体蛋白C3显著降低,犊牛免疫系统激活,系统免疫水平提高,犊牛免疫系统由Th2向Th1偏移,犊牛细胞免疫水平高于体液免疫水平;免疫应激降低了犊牛小肠粘膜SIgA水平和IEL、GC、MC数量;降低了犊牛脾脏指数和胸腺指数;免疫应激显著降低了犊牛日增重和料重比;降低了犊牛采食氮、沉积氮、氮的表观消化率、氮的表观生物学价值等指标,但未达到显著水平;显著提高了28日龄犊牛血清尿素氮含量;对犊牛腹泻率、腹泻频率等指标影响不显著。免疫应激影响了犊牛限制性氨基酸的需求比例和限制性顺序。正常条件下25~27日龄犊牛三种氨基酸的平衡比例为Lys:100、Met:29、Thr:58,53~55日龄犊牛正常组三种氨基酸的平衡比例为Lys:100、Met:30、Thr:62;而应激条件下25~27日龄犊牛三种主要限制性氨基酸的平衡比例为Lys:100、Met:27、Thr:61,53~55日龄犊牛三种限制性氨基酸的平衡比例为Lys:100、Met:27、Thr:60。并且三种氨基酸的限制性顺序发生了变化,正常时为赖氨酸、蛋氨酸、苏氨酸,应激时为赖氨酸、苏氨酸、蛋氨酸。
The neonatal calf has heightened susceptibility to infectious disease. Traditional calf-rearing programs limited nutrient intake from milk to promote dry-feed intake and early weaning. Dramatic improvements in growth performance achieved by feeding milk replacer with plant protein is hypothesized to be associated with enhanced immune function and increased resistance of the calf to infectious disease. Objectives of this dissertation were to determine the influence of protein nutrition on the immune response of the calf, and to improve the balance of amino acid and the performance of the calf.
     1. The influence of protein level on the related indexes of immune response of the calf.
     Fifteen male Chinese Holstein calves were selected to determine the influence of protein level of milk replacer on the immune response and other serum index of the calf. The results indicated that the highest IgG and AKP in serum (P>0.05), the live body weight and the index of spleen were observed when the protein level is 22% of milk replacer. And the concentrations of IL-1, the population of mast cells (P<0.05) were significantly improved with the increase of protein level of milk replacer. The GH, IGF-1, TAOC in serum had a trend of increase with higher of protein level of milk replacer, but the concentration of NO in serum decreased with highest protein level of milk replacer. The results suggest that it was beneficial to the anabolic metabolism of protein and the immune enhancement when the protein level is 22%.
     2. The influence of plant protein content on the related indexes of immune response of the calf.
     Twelve male Chinese Holstein calves were selected to determine the influence of plant protein content of milk replacer on the related indexes of immune response of the calf between 6~11days . The results indicated that the highest total protein (P>0.05), GH (P>0.05), IgG (P>0.05) and TAOC (P<0.05), NO (P<0.05), AKP (P<0.05), the lowest urea nitrogen (P>0.05) in serum of calves between 6~9 days were observed when the plant protein content is 50% in milk replacer. And the diarrhea was serious when the plant protein content is 80% of total protein of milk replacer, especially between 6~8 days.
     3. The influence of plant protein content on the related indexes of immune response of the calf.
     Twelve male Chinese Holstein calves were selected to determine the influence of plant protein content of milk replacer on the related indexes of immune response of the calf between 2~8 weeks . The results indicated that the concentration of IgG (P>0.05), IL-1 (P>0.05), TNF-α(P>0.05), AKP (P<0.05)were highest when the plant protein content is 50% of total protein in milk replacer And the concentration of IgG、IL-1 and TNF-αhad a trend of decrease between 2~4 weeks, but had a trend of increase between 4~8 weeks. The concentration of SIgA and the population of mast cells (P<0.05) in mucosal became lower and the population of goblet cells (P<0.05) in mucosal became higher with the increase of plant protein content in milk replacer. But there no influence of plant protein content of milk replacer on the concentration of lysozyme, GH and IGF-1 in serum. The morbidity of calves significantly enhanced when the plant protein content is 80% of total protein of milk replacer. The results suggest that the calf can adapt to massive plant protein after 4 weeks.
     4. The influence of main limited amino acid on the immune response and the performance of calves
     Twenty male Chinese Holstein calves were selected determine the influence of main limited amino acid (LAA) on the immune response and the performance of calves. The results indicated that the deficiency of LAA decreased the intake, BW, ADG and the nitrogen retention, digestibility and biological value of nitrogen. And the lysine had most distinct effect on the performance and the nitrogen balance, and then was the methionine and the threonine. The lysine deficiency increased the innate immunity response; however, the threonine had little influence. The lysine and methionine deficiency respectively made the immune system migration from Th1 type to Th2 type, the humeral immune response enhanced. The morbidity and the mortality of calves also improved significantly with the deficiency of lysine and methionine respectively. The threonine deficiency decreased the organ index of thymus, spleen and hepar.
     5. The influence of immunological stress on the optimum amino acid pattern of LAA of calves Forty male Chinese Holstein calves were selected to determine the influence of immunological stress on the optimum amino acid pattern of LAA of calves by the method amino acid deletion. The body temperature enhanced significantly with the challenge of lipopolysaccharide. The results indicated that the immunological stress enhanced the innate and adapt immune response, and the migration of immune response from Th2 type to Th1 type happened. But the intestinal mucosal immune response decreased. The performance and nitrogen balance also were suppressed by the immunological stress. More importantly, the limit sequence of LAA were changed from lysine > methionine > threonine to lysine > threonine > methionine, and the optimum amino acid pattern of Lys/Met/Thr for calves between 25~27 days changed from 100/29/58 to 100/27/61, the Pattern for calves between 55~57 days changed from 100/30/62 to 100/27/60 with the challenge of LPS.
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