肉仔鸡消化道应激评价及其机理研究
发布时间：2018-07-14 18:23
【摘要】：本论文以小麦面筋蛋白为基础试验材料,经过美拉德反应和氧化反应修饰为异蛋白,模拟肉仔鸡消化道应激,以此研究应激对消化道结构和功能的影响及其机理,同时初步建立应激评价体系,并通过添加抗应激剂研究缓解消化道应激的技术。试验一高级糖基化终产物诱导的肉仔鸡消化道应激本试验旨在研究面筋蛋白AGE对肉仔鸡消化道结构和功能的影响。将180只肉公鸡分为3个处理,每个处理6个重复,每重复10只鸡。分别饲喂对照日粮(CON),原始面筋日粮(WG)和面筋蛋白AGE日粮(AGE)。结果表明,AGE处理组显著降低21日龄肉仔鸡BW,ADG和ADFI,并显著增加饲料转化率(P0.05)。三个处理组中粪便pH值,粪便水分含量和回肠形态组织学并无显著差异。然而,AGE处理组的粗蛋白质表观消化率显著低于对照组的相应值(P0.05)。对35日龄肉仔鸡而言,三个处理组中AGE处理组的BW最低,而FCR值最高。结论:面筋蛋白的美拉德反应产物降低肉仔鸡的生长性能。AGE是肉仔鸡消化道的一种应激源,会影响消化道的功能而并未改变肠道形态。试验二氧化面筋诱导的肉仔鸡消化道应激本研究旨在探讨氧化面筋对肉仔鸡消化道结构和功能的影响。将180只肉公鸡随机分为3个处理组,每个处理6个重复,每个重复10只鸡。分别饲喂对照日粮(CON),原始面筋日粮(WG)和氧化面筋日粮(OG)。结果表明,氧化面筋日粮显著降低21日龄和35日龄肉仔鸡BW和ADG,并显著增加饲料转化率(P0.05)。与对照组相比,氧化面筋处理组粪便pH值显著降低(P0.05),粪便水分含量显著增加(P0.05),试验动物表现出腹泻症状。三种处理日粮对肉仔鸡回肠形态结构无显著影响。与饲喂对照组日粮相比,饲喂OG处理日粮的肉仔鸡显著降低粗蛋白质的表观消化率(P0.05)。结论:氧化面筋蛋白是肉仔鸡消化道应激源之一,造成肉仔鸡消化道处于应激状态,发生腹泻症状。氧化面筋会降低肉仔鸡的生长性能,可以诱导消化道功能的改变,但并未改变回肠组织结构。试验三肉仔鸡消化道应激生物标志物筛选本研究旨在筛选应激状态下肉仔鸡血清、嗉囊和回肠中生物标志物和组织中差异表达蛋白,用于更准确地预测或判定消化道的应激状态。将180只肉公鸡随机分为3个处理组,每个处理6个重复,每个重复10只鸡。分别饲喂对照日粮(CON),原始面筋日粮(WG)和氧化面筋日粮(OG)。结果表明,消化道应激状态下,嗉囊和回肠组织中氧化还原平衡遭破坏,并表现出一定程度的炎症免疫状态,而血清中氧化状态和炎症状态并未受到应激状态的影响。除生长性能、粪便表观状态、粪便p H值、粪便水分含量、粗蛋白质表观消化率显著变化外,消化道组织中GSH、GSSG、GSH/GSSG、LPO、MDA、IL-10和TNF-α发生显著变化。蛋白质组学分析表明,消化道应激状态下嗉囊和回肠组织中m GST和PLA2表达量下调,抗菌肽1、抗菌肽2和IL-18表达量下调。结论:反映肉仔鸡消化道应激状态的生物标志物包括,生长性能、粪便表观评分、粪便pH值、粪便初水分含量、粗蛋白质表观消化率、消化道组织中氧化状态指标和炎症免疫状态指标。依据指标数据采集测定过程对肉仔鸡损伤程度、指标测定难易程度和价格信息,将生物标志物分为两个层级。试验四肉仔鸡消化道应激机理研究本研究利用iTRAQ方法以及质谱和软件分析,探索应激状态下消化道功能改变的原因和通路。将120只肉公鸡随机分为2个处理组,每个处理6个重复,每个重复10只鸡。分别饲喂对照日粮(CON)和氧化面筋日粮(OG)。结果表明,肉仔鸡消化道组织中共鉴定出4385个蛋白质,其中对照组和应激组间共有76个蛋白差异化表达。其中53个蛋白表达量上调,23个蛋白表达量下调。KEGG代谢通路分析发现嗉囊和回肠组织中有1条代谢通路被显著性差异调控。结论:消化道应激状态下,上皮细胞的缝隙连接通路被显著性差异调控,调控细胞内水分和离子转运的功能失调,造成肉仔鸡腹泻。消化道上皮细胞收缩性的改变可能是诱导炎症反应的原因,发挥抗原递呈作用的蛋白质表达量上调,造成消化道中免疫状态激活,而具备清除过氧化物和脂质代谢产物功能的蛋白质表达量下调导致消化道中氧化还原平衡破坏。试验五维生素C对肉仔鸡消化道抗应激作用的研究本试验旨在研究维生素C缓解消化道应激状态的作用。将240只肉公鸡随机分为4个处理组,每个处理6个重复,每个重复10只鸡。分别饲喂对照日粮(CON)、面筋日粮(WG),氧化面筋日粮(OG)和OG+0.4%维生素C日粮(AOG)。结果表明,与氧化面筋组相比,添加Vc处理组显著改善肉仔鸡粪便状态,显著提高粪便pH值,降低粪便中水分含量(P0.05);同时,Vc日粮显著提高消化道应激后肉仔鸡的生长性能,和粗蛋白质表观消化率(P0.05);与氧化面筋处理组相比,AOG处理组回肠中GSSG和MDA浓度显著降低(P0.05)。结论:维生素C改善消化道应激状态下肉仔鸡消化道功能,从而缓解肉仔鸡消化道应激对生长性能的抑制作用。维生素C通过抑制消化道中的氧化应激,发挥其缓解由氧化面筋诱导的消化道应激的作用。
[Abstract]:In this paper, the effects of stress on digestive tract structure and function were studied by Maillard reaction and oxidation reaction modified as isoprotein, and the effects of stress on digestive tract structure and function were studied. At the same time, stress evaluation system was established and stress relieving agent was added to alleviate digestive stress. The test of a high glycosylation end product induced digestive tract stress in broilers was designed to study the effects of gluten AGE on the structure and function of digestive tract in broilers. 180 meat roosters were divided into 3 treatments, each of which was treated with 6 repetitions, and 10 chickens were repeated each. The control diet (CON), raw gluten diet (WG) and gluten A were fed. GE diet (AGE). The results showed that the AGE treatment group significantly reduced the 21 day old broilers' BW, ADG and ADFI, and significantly increased the feed conversion rate (P0.05). There was no significant difference in the fecal pH value in the three treatment groups, and there was no significant difference between the fecal moisture content and the intestinal morphology histology. However, the apparent digestibility of the crude protein in the AGE treatment group was significantly lower than the corresponding value of the control group (P0.05). For 35 day old broilers, the BW of the AGE treatment group in the three treatment groups was the lowest, and the FCR value was the highest. Conclusion: the mallard reaction product of gluten protein decreased the growth performance of broilers, and.AGE was a stressor in the digestive tract of broilers, which could affect the function of the digestive tract and did not change the intestinal form. Test the broilers induced by two oxidized gluten. The effects of digestive stress on the structure and function of digestive tract in broilers were investigated. 180 meat roosters were randomly divided into 3 treatment groups, each of which was treated 6 repetitions, each repeated 10 chickens. The control diet (CON), the raw gluten diet (WG) and the aerobic gluten diet (OG) were fed respectively. The results showed that the dietary gluten diet was significant Reducing the BW and ADG of 21 day and 35 day old broilers and significantly increasing the feed conversion rate (P0.05). Compared with the control group, the fecal pH value of the oxidative gluten treatment group was significantly decreased (P0.05), the water content in the feces was significantly increased (P0.05), and the experimental animals showed diarrhea symptoms. Three kinds of rations had no significant effect on the morphology and structure of the broilers. Compared with the control diet, the apparent digestibility (P0.05) of crude protein was significantly reduced in the broilers fed with OG treatment. Conclusion: the oxidative gluten protein is one of the stress sources of the digestive tract in broilers, causing the digestive tract in the broilers to be in the stress state and the diarrhea symptoms. The oxidation gluten can reduce the growth performance of Broilers and can induce the digestive tract work. The change of energy, but did not change the ileum tissue structure. Test three broiler digestive tract stress biomarkers screening this study aims to screen the serum of broilers under stress, the differentially expressed proteins in the crop and the crop and the ileum, to predict or determine the stress state of the digestive tract more accurately. 180 meat cocks are random. It was divided into 3 treatment groups, each treated with 6 repetitions, each repeated 10 chickens. The control diet (CON), the original gluten diet (WG) and the oxidized gluten diet (OG). The results showed that the redox balance in the crop and the ileum was destroyed in the digestive tract, and showed a certain degree of inflammatory immune state, and the oxidation in the serum. State and inflammatory state were not affected by stress state. Except growth performance, fecal apparent state, fecal P H value, fecal water content, and apparent protein apparent digestibility, GSH, GSSG, GSH/GSSG, LPO, MDA, IL-10 and TNF- alpha in digestive tract were significantly changed. Protein histology analysis showed that the crop under digestive tract stress state The expression of M GST and PLA2 decreased in the ileum and the expression of antibacterial peptide 1, antibacterial peptide 2 and IL-18 decreased. Conclusion: the biomarkers reflecting the stress state of digestive tract in broilers include growth performance, stool apparent score, fecal pH value, initial water content of feces, apparent digestibility of crude protein, oxidation state index and inflammation in digestive tract tissue The index of immune state. According to the index data, the degree of injury of broiler was collected and the index was measured, the index was measured and the biomarker was divided into two levels. The study on the stress mechanism of digestive tract in four broilers was studied by using iTRAQ method, mass spectrometry and software analysis to explore the changes of digestive tract function under stress condition. The 120 meat roosters were randomly divided into 2 treatment groups, each of which was treated with 6 repetitions, each repeated 10 chickens. The control diet (CON) and the oxidized gluten diet (OG) were fed respectively. The results showed that 4385 proteins were identified in the digestive tract tissues of the broilers, of which 76 protein differential expressions were found between the control group and the stress group. The expression of 53 proteins was up-regulated, and 23 protein expression levels were down regulated by.KEGG pathway analysis. 1 metabolic pathways in the crop and ileum were significantly regulated. Conclusion: the gap junction pathway of epithelial cells is regulated by significant difference under the stress state of the digestive tract, regulating the dysfunction of water and ion transport in the cells. The changes in the contractility of the epithelial cells of the digestive tract may be the cause of the inflammatory reaction, and the expression of the protein expression in the digestive tract is up-regulated, causing the activation of the immune state in the digestive tract, and the downregulation of the protein expression in the digestive tract and the function of the metabolites of the peroxide and lipid leads to the redox level in the digestive tract. Test five vitamin C on the anti stress effect of vitamin C on the digestive tract of broilers to study the effect of vitamin C on the stress state of the digestive tract. 240 meat roosters were randomly divided into 4 treatment groups, each treated with 6 repetitions, and each repeated 10 chickens. The control diet (CON), gluten diet (WG), and oxidized gluten diet (OG) were fed. And OG+0.4% vitamin C diet (AOG). The results showed that compared with the oxidative gluten group, the addition of Vc treatment group significantly improved the fecal state of broilers, significantly increased the pH value of the feces and reduced the water content in the feces (P0.05). At the same time, the Vc diet significantly improved the growth performance of Broilers and the apparent digestibility of protein (P0.05) after the digestive tract stress, and the oxidation of the crude protein (P0.05). Compared with the gluten treatment group, the concentration of GSSG and MDA in the ileum of AOG treatment group decreased significantly (P0.05). Conclusion: vitamin C improves digestive tract function of broilers under the stress of digestive tract, thus alleviates the inhibitory effect of digestive tract stress on growth performance of broilers. Vitamin C can relieve the oxidative stress by inhibiting oxidative stress in the alimentary tract and relieving the oxidative gluten. The effect of induced digestive stress.
【学位授予单位】：中国农业科学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S831.5
，
本文编号：2122569
[Abstract]:In this paper, the effects of stress on digestive tract structure and function were studied by Maillard reaction and oxidation reaction modified as isoprotein, and the effects of stress on digestive tract structure and function were studied. At the same time, stress evaluation system was established and stress relieving agent was added to alleviate digestive stress. The test of a high glycosylation end product induced digestive tract stress in broilers was designed to study the effects of gluten AGE on the structure and function of digestive tract in broilers. 180 meat roosters were divided into 3 treatments, each of which was treated with 6 repetitions, and 10 chickens were repeated each. The control diet (CON), raw gluten diet (WG) and gluten A were fed. GE diet (AGE). The results showed that the AGE treatment group significantly reduced the 21 day old broilers' BW, ADG and ADFI, and significantly increased the feed conversion rate (P0.05). There was no significant difference in the fecal pH value in the three treatment groups, and there was no significant difference between the fecal moisture content and the intestinal morphology histology. However, the apparent digestibility of the crude protein in the AGE treatment group was significantly lower than the corresponding value of the control group (P0.05). For 35 day old broilers, the BW of the AGE treatment group in the three treatment groups was the lowest, and the FCR value was the highest. Conclusion: the mallard reaction product of gluten protein decreased the growth performance of broilers, and.AGE was a stressor in the digestive tract of broilers, which could affect the function of the digestive tract and did not change the intestinal form. Test the broilers induced by two oxidized gluten. The effects of digestive stress on the structure and function of digestive tract in broilers were investigated. 180 meat roosters were randomly divided into 3 treatment groups, each of which was treated 6 repetitions, each repeated 10 chickens. The control diet (CON), the raw gluten diet (WG) and the aerobic gluten diet (OG) were fed respectively. The results showed that the dietary gluten diet was significant Reducing the BW and ADG of 21 day and 35 day old broilers and significantly increasing the feed conversion rate (P0.05). Compared with the control group, the fecal pH value of the oxidative gluten treatment group was significantly decreased (P0.05), the water content in the feces was significantly increased (P0.05), and the experimental animals showed diarrhea symptoms. Three kinds of rations had no significant effect on the morphology and structure of the broilers. Compared with the control diet, the apparent digestibility (P0.05) of crude protein was significantly reduced in the broilers fed with OG treatment. Conclusion: the oxidative gluten protein is one of the stress sources of the digestive tract in broilers, causing the digestive tract in the broilers to be in the stress state and the diarrhea symptoms. The oxidation gluten can reduce the growth performance of Broilers and can induce the digestive tract work. The change of energy, but did not change the ileum tissue structure. Test three broiler digestive tract stress biomarkers screening this study aims to screen the serum of broilers under stress, the differentially expressed proteins in the crop and the crop and the ileum, to predict or determine the stress state of the digestive tract more accurately. 180 meat cocks are random. It was divided into 3 treatment groups, each treated with 6 repetitions, each repeated 10 chickens. The control diet (CON), the original gluten diet (WG) and the oxidized gluten diet (OG). The results showed that the redox balance in the crop and the ileum was destroyed in the digestive tract, and showed a certain degree of inflammatory immune state, and the oxidation in the serum. State and inflammatory state were not affected by stress state. Except growth performance, fecal apparent state, fecal P H value, fecal water content, and apparent protein apparent digestibility, GSH, GSSG, GSH/GSSG, LPO, MDA, IL-10 and TNF- alpha in digestive tract were significantly changed. Protein histology analysis showed that the crop under digestive tract stress state The expression of M GST and PLA2 decreased in the ileum and the expression of antibacterial peptide 1, antibacterial peptide 2 and IL-18 decreased. Conclusion: the biomarkers reflecting the stress state of digestive tract in broilers include growth performance, stool apparent score, fecal pH value, initial water content of feces, apparent digestibility of crude protein, oxidation state index and inflammation in digestive tract tissue The index of immune state. According to the index data, the degree of injury of broiler was collected and the index was measured, the index was measured and the biomarker was divided into two levels. The study on the stress mechanism of digestive tract in four broilers was studied by using iTRAQ method, mass spectrometry and software analysis to explore the changes of digestive tract function under stress condition. The 120 meat roosters were randomly divided into 2 treatment groups, each of which was treated with 6 repetitions, each repeated 10 chickens. The control diet (CON) and the oxidized gluten diet (OG) were fed respectively. The results showed that 4385 proteins were identified in the digestive tract tissues of the broilers, of which 76 protein differential expressions were found between the control group and the stress group. The expression of 53 proteins was up-regulated, and 23 protein expression levels were down regulated by.KEGG pathway analysis. 1 metabolic pathways in the crop and ileum were significantly regulated. Conclusion: the gap junction pathway of epithelial cells is regulated by significant difference under the stress state of the digestive tract, regulating the dysfunction of water and ion transport in the cells. The changes in the contractility of the epithelial cells of the digestive tract may be the cause of the inflammatory reaction, and the expression of the protein expression in the digestive tract is up-regulated, causing the activation of the immune state in the digestive tract, and the downregulation of the protein expression in the digestive tract and the function of the metabolites of the peroxide and lipid leads to the redox level in the digestive tract. Test five vitamin C on the anti stress effect of vitamin C on the digestive tract of broilers to study the effect of vitamin C on the stress state of the digestive tract. 240 meat roosters were randomly divided into 4 treatment groups, each treated with 6 repetitions, and each repeated 10 chickens. The control diet (CON), gluten diet (WG), and oxidized gluten diet (OG) were fed. And OG+0.4% vitamin C diet (AOG). The results showed that compared with the oxidative gluten group, the addition of Vc treatment group significantly improved the fecal state of broilers, significantly increased the pH value of the feces and reduced the water content in the feces (P0.05). At the same time, the Vc diet significantly improved the growth performance of Broilers and the apparent digestibility of protein (P0.05) after the digestive tract stress, and the oxidation of the crude protein (P0.05). Compared with the gluten treatment group, the concentration of GSSG and MDA in the ileum of AOG treatment group decreased significantly (P0.05). Conclusion: vitamin C improves digestive tract function of broilers under the stress of digestive tract, thus alleviates the inhibitory effect of digestive tract stress on growth performance of broilers. Vitamin C can relieve the oxidative stress by inhibiting oxidative stress in the alimentary tract and relieving the oxidative gluten. The effect of induced digestive stress.
【学位授予单位】：中国农业科学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S831.5
，
本文编号：2122569
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