【摘要】：禽類產(chǎn)品是我國人民肉類消費的重要來源。近30年來,我國居民對禽產(chǎn)品的消費需求不斷增加。但是受國內養(yǎng)殖業(yè)技術水平的限制,家禽養(yǎng)殖常常遭受疾病、營養(yǎng)失衡、應激等影響,使得禽產(chǎn)品供應經(jīng)常發(fā)生較大波動。因此,如何提高家禽健康水平,提升家禽生產(chǎn)成績,保障禽產(chǎn)品穩(wěn)定供應以滿足居民需求是我國農(nóng)業(yè)產(chǎn)業(yè)的重要課題。轉移因子(Transfer factor, TF)是一組由T淋巴細胞釋放的沒有抗原性的小分子多肽、核苷酸、氨基酸類物質組成的混合液,近年來作為增強免疫制劑在家禽生產(chǎn)領域的應用引起了越來越多人的關注。本試驗對出殼后5日齡雛雞灌服0.1mg、0.25mg及1.0mg劑量的雞脾轉移因子,對照組灌服生理鹽水,至40日齡時取十二指腸、空腸、回腸及直腸組織,從腸道黏膜組織學結構、細胞因子、MUC2(黏蛋白基因2)及TLR2/4 (Toll樣受體2/4)表達及腸道抗氧化功能等途徑研究了雞脾轉移因子的作用機制。試驗同時研究了雞脾轉移因子在生產(chǎn)中的應用,分析了其對雞群增重、免疫器官發(fā)育和新城疫治療效果的影響。結果如下：1.雞脾轉移因子對雞腸黏膜組織學結構影響1.0mg劑量雞脾轉移因子可以顯著增加40日齡試驗雞小腸的絨毛高度、腸絨毛高度／隱窩深度比值(V/C)(P0.05)和杯狀細胞密度(P0.05)；但各劑量雞脾轉移因子對直腸杯狀細胞數(shù)目的影響與對照組相比較差異不顯著。結果表明雞脾轉移因子可改善雞小腸的黏膜屏障功能。2.雞脾轉移因子對雞腸道細胞因子的影響1.0mg劑量的雞脾轉移因子顯著提高40日齡雞十二指腸、空腸、回腸和直腸的IL-10含量(P0.05)。0.25mmg劑量雞脾轉移因子顯著提高試驗組空腸、回腸、直腸IL-13含量(P0.05)。O.1mg和0.25mg劑量的雞脾轉移因子顯著降低40日齡雞十二指腸的IL-1水平(P0.05)。0.25mg和1.0mg劑量水平的雞脾轉移因子顯著降低試驗組十二指腸內TNF-α表達水平(P0.05)。結果表明雞脾轉移因子可提高雞腸道抗炎因子表達和抑制促炎因子分泌,改善腸道黏膜免疫屏障功能。3.雞脾轉移因子對雞腸道MUC2、TLR2及TLR4 mRNA表達的影響灌服1.0mmg劑量的雞脾轉移因子可顯著增加40日齡試驗雞十二指腸、回腸、直腸MUC2 mRNA水平(P0.05)。灌服0.25mg和1.0mg劑量雞脾轉移因子均可以顯著降低TLR2、TLR4基因在十二指腸(P0.05)、空腸(P0.05)、回腸(P0.01；P0.05)和直腸(P0.01；P0.05)的表達水平,且這種作用在一定程度上表現(xiàn)出劑量依賴性。4.雞脾轉移因子對雞腸道抗氧化功能的影響灌服0.25mg雞脾轉移因子顯著提高40日齡雞十二指腸、空腸、直腸CAT活性(P0.05)。0.25mg劑量組的十二指腸、空腸GSH-Px活力與對照組相比顯著升高(P0.05)。0.25mg和1.0mg劑量的雞脾轉移因子可顯著降低十二指腸、空腸及回腸內MDA含量(P0.05)。0.25mg劑量的雞脾轉移因子可顯著性提高十二指腸、回腸、直腸的T-AOC(P0.05)。結果表明雞脾轉移因子可提高雞腸道組織的抗氧化功能。5.雞脾轉移因子對免疫器官發(fā)育及疾病抵抗能力的影響對5日齡雛雞隔周灌服1.0mg劑量的雞脾轉移因子,40日齡試驗組雞體重顯著高于對照組(P0.05)。雛雞5日齡起每日灌服0.25mg劑量的雞脾轉移因子可以顯著提高14日齡和22日齡脾臟與胸腺重量,極顯著提高新城疫免疫后的抗體效價(P0.001)。6.雞脾轉移因子在養(yǎng)雞生產(chǎn)中的應用大群試驗結果顯示雞脾轉移因子可有效提高規(guī)�；a(chǎn)雞群的成活率、個體體重,降低料肉比,提高養(yǎng)殖戶的綜合效益。對感染新城疫的生產(chǎn)雞群應急處理顯示,雞脾轉移因子可以顯著提高雞群的抵抗力,降低感染新城疫雞群的死亡率,大大降低因疫病傳播帶來的損失。結論：雞脾轉移因子可以顯著提高雞小腸絨毛高度,增加雞小腸V/C比值,誘導腸道干細胞增殖分化發(fā)育,增加杯狀細胞密度,改善腸道黏膜機械屏障功能；改變腸道細胞因子的動態(tài)分布,促進腸道抗炎因子IL-10及IL-13的表達,降低促炎因子IL-1和TNF-α水平,改善腸道黏膜免疫屏障功能,進而提高雞群抗病能力和生長性能。其作用機制可能與雞脾轉移因子提高腸道MUC2基因的表達水平、下調各個腸段TLR2及TLR4 mRNA水平和提高腸道抗氧化功能有關。
[Abstract]:Poultry products are an important source of meat consumption in our country. In the past 30 years, the demand for consumption of poultry products has increased in our country. However, by the limitation of the technical level of domestic industry, poultry breeding often suffers from diseases, nutrition imbalance, stress and so on, so that the supply of poultry products often fluctuates greatly. Therefore, how to improve the health level of the poultry, to improve the performance of the poultry production, to guarantee the stable supply of the poultry products to meet the needs of the residents is an important subject of the agricultural industry in China. Transfer factor (TF) is a group of mixed liquor consisting of small, non-antigenic, small-molecular, nuclear, and amino-acid-like substances released by T-lymphocytes, which has attracted more and more attention in recent years as an increase in the application of immune preparation in the field of poultry production. In the experiment, the chicken spleen transfer factor of 0.1 mg, 0.25 mg and 1.0 mg was given to the five-day-old chicks after the shell, and the control group was given physiological saline, and the duodenum, jejunum, ileum and rectum were taken from the intestinal mucosa, and the histological structure and the cytokines of the intestinal mucosa were taken from the intestinal mucosa. MUC2 (mucin gene 2) and TLR2/4 (Toll-like receptor 2/4) expression and intestinal anti-oxidation function were used to study the mechanism of chicken spleen transfer factor. The application of chicken spleen transfer factor in production was also studied, and its effects on the weight gain, immune organ development and the treatment effect of Newcastle disease were analyzed. The results are as follows:1. The effect of the chicken's spleen transfer factor on the histological structure of the chicken's intestinal mucosa was 1.0 mg, and the spleen transfer factor of the chicken was significantly increased by 40-day-old, the ratio of the depth of the intestinal villus to the depth of the crypt (V/ C) (P0.05) and the density of the cup-like cells (P0.05). However, the effect of each dose of chicken spleen transfer factor on the number of the rectal goblet cells was not significantly different from that in the control group. The results show that the chicken spleen transfer factor can improve the mucosal barrier function of the small intestine of the chicken. The effect of the chicken's spleen transfer factor on the intestinal cytokines of the chicken was higher than that of the chicken's spleen transfer factor of 1.0 mg, and the content of IL-10 in the duodenum, jejunum, ileum and rectum of the 40-day-old chicken was significantly increased (P0.05). The spleen transfer factor of the 0.25 mmg dose of chicken was significantly increased in the jejunum and ileum of the experimental group. The level of IL-1 in the duodenum of the 40-day-old chicken was significantly lower than that of the chicken's spleen transfer factor (P <0.05). The level of TNF-1 in the duodenum of the test group was significantly reduced by the chicken spleen transfer factor of 0.25 mg and 1.0 mg (P0.05). The results show that the chicken spleen transfer factor can improve the anti-inflammatory factor expression of the chicken intestinal tract and inhibit the secretion of the pro-inflammatory factor and improve the immune barrier function of the intestinal mucosa. The effect of chicken spleen transfer factor on the expression of MUC2, TLR2 and TLR4 mRNA in chicken's intestinal tract significantly increased the level of MUC2 mRNA in the duodenum, ileum and rectum of the 40-day-old test (P0.05). The expression level of TLR2 and TLR4 gene in the duodenum (P0.05), jejunum (P0.05), ileum (P0.01), and rectum (P0.01) and rectum (P0.01) and rectum (P0.01) was significantly reduced. The effect of the chicken's spleen transfer factor on the anti-oxidation of the chicken's intestinal tract was significantly increased in the duodenum, jejunum and rectum of the 40-day-old chicken (P0.05). The duodenum of the 0.25 mg dose group, The activity of GSH-Px in the jejunum was significantly higher than that in the control group (P0.05). The dose of 0.25 mg and 1.0 mg of the chicken spleen transfer factor could significantly decrease the content of MDA in the duodenum, jejunum and ileum (P0.05). The dose of 0.25 mg of the chicken spleen transfer factor could significantly increase the T-AOC of the duodenum, the ileum and the rectum (P0.05). The results show that the chicken's spleen transfer factor can improve the anti-oxidation function of the chicken's intestinal tissue. The effect of the chicken's spleen transfer factor on the development of immune organs and the resistance of the disease to the disease resistance was significantly higher than that of the control group (P0.05). The spleen and thymus weight of the spleen and thymus of the 14-day-old and 22-day-old chickens were significantly improved, and the antibody titer after the immunization of the newcastle disease was significantly improved (P 0.001). The results show that the chicken spleen transfer factor can effectively improve the survival rate of the chicken group, the individual weight, the ratio of the feed and the meat, and improve the comprehensive benefit of the farmers. The chicken-spleen transfer factor can obviously improve the resistance of the chicken group, reduce the mortality of the chicken group infected with the newcastle disease, and greatly reduce the loss caused by the transmission of the disease, according to the emergency treatment of the chicken group infected with the newcastle disease. Conclusion: The chicken spleen transfer factor can significantly improve the height of the small intestine of the chicken, increase the V/ C ratio of the small intestine of the chicken, induce the proliferation and differentiation of the intestinal stem cells, increase the density of the goblet cells, improve the mechanical barrier function of the intestinal mucosa, and change the dynamic distribution of the intestinal cytokines, The expression of IL-10 and IL-13 in the intestinal tract is promoted, the IL-1 and TNF-1 levels of the proinflammatory factors are reduced, the immune barrier function of the intestinal mucosa is improved, and the disease resistance and the growth performance of the chicken group are further improved. The mechanism of action may be related to the improvement of the expression level of the intestinal MUC2 gene, the down-regulation of the levels of TLR2 and TLR4 mRNA in the intestinal segments and the improvement of the anti-oxidation function of the intestinal tract.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S831

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