【摘要】：本論文以小麥面筋蛋白為基礎試驗材料,經(jīng)過美拉德反應和氧化反應修飾為異蛋白,模擬肉仔雞消化道應激,以此研究應激對消化道結構和功能的影響及其機理,同時初步建立應激評價體系,并通過添加抗應激劑研究緩解消化道應激的技術。試驗一高級糖基化終產(chǎn)物誘導的肉仔雞消化道應激本試驗旨在研究面筋蛋白AGE對肉仔雞消化道結構和功能的影響。將180只肉公雞分為3個處理,每個處理6個重復,每重復10只雞。分別飼喂對照日糧(CON),原始面筋日糧(WG)和面筋蛋白AGE日糧(AGE)。結果表明,AGE處理組顯著降低21日齡肉仔雞BW,ADG和ADFI,并顯著增加飼料轉化率(P0.05)。三個處理組中糞便pH值,糞便水分含量和回腸形態(tài)組織學并無顯著差異。然而,AGE處理組的粗蛋白質表觀消化率顯著低于對照組的相應值(P0.05)。對35日齡肉仔雞而言,三個處理組中AGE處理組的BW最低,而FCR值最高。結論:面筋蛋白的美拉德反應產(chǎn)物降低肉仔雞的生長性能。AGE是肉仔雞消化道的一種應激源,會影響消化道的功能而并未改變腸道形態(tài)。試驗二氧化面筋誘導的肉仔雞消化道應激本研究旨在探討氧化面筋對肉仔雞消化道結構和功能的影響。將180只肉公雞隨機分為3個處理組,每個處理6個重復,每個重復10只雞。分別飼喂對照日糧(CON),原始面筋日糧(WG)和氧化面筋日糧(OG)。結果表明,氧化面筋日糧顯著降低21日齡和35日齡肉仔雞BW和ADG,并顯著增加飼料轉化率(P0.05)。與對照組相比,氧化面筋處理組糞便pH值顯著降低(P0.05),糞便水分含量顯著增加(P0.05),試驗動物表現(xiàn)出腹瀉癥狀。三種處理日糧對肉仔雞回腸形態(tài)結構無顯著影響。與飼喂對照組日糧相比,飼喂OG處理日糧的肉仔雞顯著降低粗蛋白質的表觀消化率(P0.05)。結論:氧化面筋蛋白是肉仔雞消化道應激源之一,造成肉仔雞消化道處于應激狀態(tài),發(fā)生腹瀉癥狀。氧化面筋會降低肉仔雞的生長性能,可以誘導消化道功能的改變,但并未改變回腸組織結構。試驗三肉仔雞消化道應激生物標志物篩選本研究旨在篩選應激狀態(tài)下肉仔雞血清、嗉囊和回腸中生物標志物和組織中差異表達蛋白,用于更準確地預測或判定消化道的應激狀態(tài)。將180只肉公雞隨機分為3個處理組,每個處理6個重復,每個重復10只雞。分別飼喂對照日糧(CON),原始面筋日糧(WG)和氧化面筋日糧(OG)。結果表明,消化道應激狀態(tài)下,嗉囊和回腸組織中氧化還原平衡遭破壞,并表現(xiàn)出一定程度的炎癥免疫狀態(tài),而血清中氧化狀態(tài)和炎癥狀態(tài)并未受到應激狀態(tài)的影響。除生長性能、糞便表觀狀態(tài)、糞便p H值、糞便水分含量、粗蛋白質表觀消化率顯著變化外,消化道組織中GSH、GSSG、GSH/GSSG、LPO、MDA、IL-10和TNF-α發(fā)生顯著變化。蛋白質組學分析表明,消化道應激狀態(tài)下嗉囊和回腸組織中m GST和PLA2表達量下調,抗菌肽1、抗菌肽2和IL-18表達量下調。結論:反映肉仔雞消化道應激狀態(tài)的生物標志物包括,生長性能、糞便表觀評分、糞便pH值、糞便初水分含量、粗蛋白質表觀消化率、消化道組織中氧化狀態(tài)指標和炎癥免疫狀態(tài)指標。依據(jù)指標數(shù)據(jù)采集測定過程對肉仔雞損傷程度、指標測定難易程度和價格信息,將生物標志物分為兩個層級。試驗四肉仔雞消化道應激機理研究本研究利用iTRAQ方法以及質譜和軟件分析,探索應激狀態(tài)下消化道功能改變的原因和通路。將120只肉公雞隨機分為2個處理組,每個處理6個重復,每個重復10只雞。分別飼喂對照日糧(CON)和氧化面筋日糧(OG)。結果表明,肉仔雞消化道組織中共鑒定出4385個蛋白質,其中對照組和應激組間共有76個蛋白差異化表達。其中53個蛋白表達量上調,23個蛋白表達量下調。KEGG代謝通路分析發(fā)現(xiàn)嗉囊和回腸組織中有1條代謝通路被顯著性差異調控。結論:消化道應激狀態(tài)下,上皮細胞的縫隙連接通路被顯著性差異調控,調控細胞內水分和離子轉運的功能失調,造成肉仔雞腹瀉。消化道上皮細胞收縮性的改變可能是誘導炎癥反應的原因,發(fā)揮抗原遞呈作用的蛋白質表達量上調,造成消化道中免疫狀態(tài)激活,而具備清除過氧化物和脂質代謝產(chǎn)物功能的蛋白質表達量下調導致消化道中氧化還原平衡破壞。試驗五維生素C對肉仔雞消化道抗應激作用的研究本試驗旨在研究維生素C緩解消化道應激狀態(tài)的作用。將240只肉公雞隨機分為4個處理組,每個處理6個重復,每個重復10只雞。分別飼喂對照日糧(CON)、面筋日糧(WG),氧化面筋日糧(OG)和OG+0.4%維生素C日糧(AOG)。結果表明,與氧化面筋組相比,添加Vc處理組顯著改善肉仔雞糞便狀態(tài),顯著提高糞便pH值,降低糞便中水分含量(P0.05);同時,Vc日糧顯著提高消化道應激后肉仔雞的生長性能,和粗蛋白質表觀消化率(P0.05);與氧化面筋處理組相比,AOG處理組回腸中GSSG和MDA濃度顯著降低(P0.05)。結論:維生素C改善消化道應激狀態(tài)下肉仔雞消化道功能,從而緩解肉仔雞消化道應激對生長性能的抑制作用。維生素C通過抑制消化道中的氧化應激,發(fā)揮其緩解由氧化面筋誘導的消化道應激的作用。
[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.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S831.5
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本文編號：2122569