百里香酚和香芹酚对肉仔鸡肠上皮屏障和免疫功能的调节作用

发布时间：2018-06-11 16:02

  本文选题：植物精油 + 肉仔鸡　； 参考：《中国农业大学》2016年博士论文

【摘要】：本论文通过四个动物试验研究了日粮中添加以百里香酚和香芹酚为活性成分的植物精油(EO)对肉仔鸡肠上皮屏障功能和免疫功能的调节作用。试验一通过2×2因子完全随机设计研究了日粮中添加EO(活性成分为百里香酚和香芹酚)对脂多糖(LPS)诱导的肉仔鸡炎症反应的调节作用。试验结果表明,日粮中添加EO不影响LPS注射期间的肉仔鸡生产性能以及器官相对重量,但显著降低了LPS注射后3 h和24 h肝脏MDA的含量(P0.05)。同时,在LPS注射后3 h,EO显著上调了脾脏IL-1β和iNOS的mRNA表达量而下调了TLR4的表达量(P0.05)：在注射后8 h,EO显著上调了IL-4的表达量而下调了TNF-a和TLR4的表达量(P0.05),说明EO可提高肝脏的抗氧化力,并且对脾脏炎症相关基因的表达具有显著的调节作用。试验二通过2×2因子完全随机设计研究了日粮中添加EO对球虫感染所造成的肉仔鸡肠道损伤的调节作用。试验结果表明,球虫感染后一周,肉仔鸡体增重和饲料转化率显著降低、小肠组织形态明显受损(P0.05),小肠紧密连接蛋白occludin的mRNA表达量显著下调,而IFN-γ和IL-Ip的mRNA表达量显著上调(P0.05)。日粮中EO的添加降低了球虫感染肉仔鸡的粪便带血程度和粪便中的卵囊排出量(P0.05),上调了小肠IL-4的mRNA表达(P0.05),但是不影响肉仔鸡生产性能、小肠黏膜二糖酶活力和小肠组织形态,说明EO不能有效缓解球虫感染所引起的肉仔鸡肠道结构与功能的损伤。试验三首先通过体外抑菌试验表明百里香酚和香芹酚对致病性大肠杆菌、产气荚膜梭菌和沙门氏菌有显著的抑制效果,而对有益的乳杆菌抑制效果较弱,并且百里香酚和香芹酚的抑菌效果具有相加作用。然后通过2×4因子完全随机设计研究了EO添加水平(0、60、120和240 mg/kg)对产气荚膜梭菌感染所造成的肉仔鸡肠道损伤的影响。试验结果表明,在攻毒组肉仔鸡口粮中添加EO可线性降低肠道损伤评分(P0.05),提高回肠绒毛隐窝比值(P0.05),并且120和240mg/kg的EO可显著提高血清新城疫和禽法氏囊病抗体滴度(P0.05)。随着日粮EO添加水平的升高,肉仔鸡饲料转化率有线性升高的趋势(P0.10),而回肠TLR2和TNF-a的mRNA表达量以及回肠埃希菌的数量线性降低(P0.05),此外60和120 mg/kg的EO可显著提高盲肠乳杆菌的数量(P0.05),说明EO可能通过影响肠道微生物数量、抑制TLR2介导的炎症反应来缓解产气荚膜梭菌对肉仔鸡造成的肠道损伤。试验四首先通过体外抑菌试验表明百里香酚、香芹酚及溶菌酶均能抑制产气荚膜梭菌,并且溶菌酶与百里香酚和香芹酚的抑菌效果有相加作用。在动物饲养试验中,将504只肉仔鸡随机分为9个处理,其中阴性对照组和阳性对照组饲喂小麦型基础日粮,其他处理组分别添加60、120或240 mg/kg的EO,或单独添加100 mg/kg的溶菌酶,或同时添加溶菌酶与不同剂量的EO的混合物。从d 14开始,除阴性对照组外,所有肉仔鸡每天口腔灌服1mL产气荚膜梭菌培养液,连续灌服一周：结果表明，日粮中单独添加溶菌酶或60/120 mg/kg的EO可显著降低肉仔鸡的死亡率(P0.05),降低d 21肝脏细菌移位数(P0.05)。并且,单独添加溶菌酶或EO可不同程度地减轻d 21的肠道损伤评分,改善小肠组织形态，提高肠黏膜二糖酶活力和血清溶菌酶活力。其中,日粮中单独添加120 mg/kg的EO显著提高了小肠iNOS、 occludin和claudin l的mRNA表达(P0.05)。 16s rRNA基因测序和宏基因组测序结果表明120 mg/kg EO的添加改变了回肠菌群结构和微生物功能基因的组成,提高了梭菌目和棒状杆菌属细菌的相对含量(P0.05),抑制了某些致病菌的毒力因子基因的相对丰度(P0.05)。整体来看,日粮中单独添加120 mg/kg EO对肉仔鸡的改善效果最好,而本试验所用的溶菌酶与EO联合使用不能增强其改善坏死性肠炎的作用效果。以上研究表明,日粮中添加以百里香酚和香芹酚为活性成分的EO具有显著的免疫调节作用,能够直接影响LPS诱导的肉仔鸡炎症反应,并且能够影响肠道微生物区系,改善肠上皮屏障功能,缓解产气荚膜梭菌诱导的肉仔鸡坏死性肠炎。但是,EO不能有效缓解球虫感染所造成的肠道损伤,本试验所用的外源溶菌酶与EO配伍使用不能增强EO对坏死性肠炎的改善作用。
[Abstract]:In this paper, four animal experiments were carried out to study the effect of plant essential oil (EO) adding parsley and parsley on the intestinal barrier function and immune function of chicken chick. The experiment was carried out to study the addition of EO (active ingredients for paranolol and carenol) to the diet by the complete random design of 2 x 2 factor. The experimental results showed that EO did not affect the performance of Broilers and the relative organ weight of broilers during LPS injection, but the content of MDA in 3 h and 24 h after LPS injection was significantly reduced (P0.05). Meanwhile, 3 h after LPS injection, EO significantly increased the spleen IL-1 beta and the spleen The expression of NA decreased the expression of TLR4 (P0.05): 8 h after injection, EO significantly up-regulated the expression of IL-4 and reduced the expression of TNF-a and TLR4 (P0.05), indicating that EO can improve the antioxidant capacity of the liver, and has a significant role in regulating the expression of spleen inflammation related genes. Test two was completely randomized by 2 x 2 factor design research. The results showed that the weight gain and feed conversion rate of the broilers were significantly reduced, the morphology of the small intestine was significantly impaired (P0.05), and the mRNA expression of the compact connexin occludin in the small intestine was significantly reduced, while the mRN of the small intestine compact connexin occludin was significantly down, while the mRN of IFN- gamma and IL-Ip was mRN in the week after coccidian infection. The expression of A was significantly up (P0.05). The addition of EO in the diet decreased the level of fecal blood and the excretion of oocyst in feces (P0.05), up the mRNA expression of IL-4 in the small intestine (P0.05), but did not affect the production performance of broilers, the activity of two enzymes and the morphology of small intestine in small intestinal mucosa, indicating that EO could not effectively alleviate coccidiosis. Test three the inhibitory effects of thyme and carcanolol on pathogenic Escherichia coli, Clostridium perfringens and Salmonella, and the inhibitory effect on the beneficial lactobacillus, and the inhibitory effect of thyme and carcanolol on the bacteriostasis in vitro. The effects of the addition level of EO (0,60120 and 240 mg/kg) on intestinal tract injury caused by Clostridium perfringens infection were investigated by the 2 x 4 factor complete random design. The results showed that the addition of EO in the broiler diet in the attack group could reduce the intestinal tract injury score (P0.05) and improve the ileum crypt ratio. Value (P0.05), and EO of 120 and 240mg/kg significantly increased the antibody titer of serum Newcastle disease and avian bursal disease (P0.05). With the increase of dietary EO levels, the feed conversion rate of broilers had a linear increase (P0.10), while the mRNA expression of the ileum TLR2 and TNF-a and the number of ileum in the ileum decreased linearly (P0.05), in addition to the 60 and 120 m. G/kg's EO can significantly increase the number of Lactobacillus cecium (P0.05), indicating that EO may reduce the intestinal damage caused by Clostridium perfringens by affecting the number of intestinal microbes and inhibiting the TLR2 mediated inflammatory reaction. Test four first through in vitro bacteriostasis test, the inhibition of pericarphenol, carcanolol and lysozyme could inhibit the pericarp production. Clostridium, and lysozyme had an additive effect on the bacteriostasis effect of pericolol and caramolol. In the animal feeding test, 504 broilers were randomly divided into 9 treatments, of which the negative control group and the positive control group were fed with wheat based diet, the other treatment groups added 60120 or 240 mg/kg EO, or separately added 100 mg/kg. Enzyme, or at the same time adding a mixture of lysozyme and different doses of EO. From d 14, except for negative control group, all broilers were filled with 1mL producing Clostridium perfringens culture every day for a week. The results showed that the mortality of broilers (P0.05) and D 21 were significantly reduced by adding lysozyme or 60/120 mg/kg EO in the diet. The liver bacterial migration number (P0.05). Moreover, adding lysozyme or EO alone could reduce the intestinal damage score of D 21 to varying degrees, improve the morphology of the intestinal tissue, improve the activity of two enzyme and the activity of serum lysozyme in the intestinal mucosa. Among them, the EO of 120 mg/kg in the diet was significantly higher than the iNOS, occludin and claudin L mRNA expression (P0.0). 5). The results of 16S rRNA gene sequencing and macrogenome sequencing showed that the addition of 120 mg/kg EO changed the composition of the intestinal flora structure and the microbial functional genes, increased the relative content of Clostridium and Corynebacterium bacteria (P0.05), and inhibited the relative abundance of the virulence factor genes of some pathogenic bacteria (P0.05). The addition of 120 mg/kg EO to broilers has the best effect, but the combination of lysozyme and EO used in this experiment can not enhance its effect on the improvement of necrotic enteritis. The above study shows that the dietary EO with pariolol and parsley as active ingredient has a significant immunomodulatory effect, which can directly affect the induced LPS. The inflammatory response of broilers can affect the intestinal microflora, improve the intestinal barrier function and alleviate the bad enteritis induced by Clostridium perfringens. However, EO can not effectively relieve the intestinal damage caused by coccidiosis. The combination of exogenous lysozyme and EO in this test can not enhance EO to necrotic enteritis. Improvement.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S831.5
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本文编号：2005865