本文選題：植物精油 + 肉仔雞��； 參考：《中國(guó)農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：本論文通過(guò)四個(gè)動(dòng)物試驗(yàn)研究了日糧中添加以百里香酚和香芹酚為活性成分的植物精油(EO)對(duì)肉仔雞腸上皮屏障功能和免疫功能的調(diào)節(jié)作用。試驗(yàn)一通過(guò)2×2因子完全隨機(jī)設(shè)計(jì)研究了日糧中添加EO(活性成分為百里香酚和香芹酚)對(duì)脂多糖(LPS)誘導(dǎo)的肉仔雞炎癥反應(yīng)的調(diào)節(jié)作用。試驗(yàn)結(jié)果表明,日糧中添加EO不影響LPS注射期間的肉仔雞生產(chǎn)性能以及器官相對(duì)重量,但顯著降低了LPS注射后3 h和24 h肝臟MDA的含量(P0.05)。同時(shí),在LPS注射后3 h,EO顯著上調(diào)了脾臟IL-1β和iNOS的mRNA表達(dá)量而下調(diào)了TLR4的表達(dá)量(P0.05)：在注射后8 h,EO顯著上調(diào)了IL-4的表達(dá)量而下調(diào)了TNF-a和TLR4的表達(dá)量(P0.05),說(shuō)明EO可提高肝臟的抗氧化力,并且對(duì)脾臟炎癥相關(guān)基因的表達(dá)具有顯著的調(diào)節(jié)作用。試驗(yàn)二通過(guò)2×2因子完全隨機(jī)設(shè)計(jì)研究了日糧中添加EO對(duì)球蟲感染所造成的肉仔雞腸道損傷的調(diào)節(jié)作用。試驗(yàn)結(jié)果表明,球蟲感染后一周,肉仔雞體增重和飼料轉(zhuǎn)化率顯著降低、小腸組織形態(tài)明顯受損(P0.05),小腸緊密連接蛋白o(hù)ccludin的mRNA表達(dá)量顯著下調(diào),而IFN-γ和IL-Ip的mRNA表達(dá)量顯著上調(diào)(P0.05)。日糧中EO的添加降低了球蟲感染肉仔雞的糞便帶血程度和糞便中的卵囊排出量(P0.05),上調(diào)了小腸IL-4的mRNA表達(dá)(P0.05),但是不影響肉仔雞生產(chǎn)性能、小腸黏膜二糖酶活力和小腸組織形態(tài),說(shuō)明EO不能有效緩解球蟲感染所引起的肉仔雞腸道結(jié)構(gòu)與功能的損傷。試驗(yàn)三首先通過(guò)體外抑菌試驗(yàn)表明百里香酚和香芹酚對(duì)致病性大腸桿菌、產(chǎn)氣莢膜梭菌和沙門氏菌有顯著的抑制效果,而對(duì)有益的乳桿菌抑制效果較弱,并且百里香酚和香芹酚的抑菌效果具有相加作用。然后通過(guò)2×4因子完全隨機(jī)設(shè)計(jì)研究了EO添加水平(0、60、120和240 mg/kg)對(duì)產(chǎn)氣莢膜梭菌感染所造成的肉仔雞腸道損傷的影響。試驗(yàn)結(jié)果表明,在攻毒組肉仔雞口糧中添加EO可線性降低腸道損傷評(píng)分(P0.05),提高回腸絨毛隱窩比值(P0.05),并且120和240mg/kg的EO可顯著提高血清新城疫和禽法氏囊病抗體滴度(P0.05)。隨著日糧EO添加水平的升高,肉仔雞飼料轉(zhuǎn)化率有線性升高的趨勢(shì)(P0.10),而回腸TLR2和TNF-a的mRNA表達(dá)量以及回腸埃希菌的數(shù)量線性降低(P0.05),此外60和120 mg/kg的EO可顯著提高盲腸乳桿菌的數(shù)量(P0.05),說(shuō)明EO可能通過(guò)影響腸道微生物數(shù)量、抑制TLR2介導(dǎo)的炎癥反應(yīng)來(lái)緩解產(chǎn)氣莢膜梭菌對(duì)肉仔雞造成的腸道損傷。試驗(yàn)四首先通過(guò)體外抑菌試驗(yàn)表明百里香酚、香芹酚及溶菌酶均能抑制產(chǎn)氣莢膜梭菌,并且溶菌酶與百里香酚和香芹酚的抑菌效果有相加作用。在動(dòng)物飼養(yǎng)試驗(yàn)中,將504只肉仔雞隨機(jī)分為9個(gè)處理,其中陰性對(duì)照組和陽(yáng)性對(duì)照組飼喂小麥型基礎(chǔ)日糧,其他處理組分別添加60、120或240 mg/kg的EO,或單獨(dú)添加100 mg/kg的溶菌酶,或同時(shí)添加溶菌酶與不同劑量的EO的混合物。從d 14開(kāi)始,除陰性對(duì)照組外,所有肉仔雞每天口腔灌服1mL產(chǎn)氣莢膜梭菌培養(yǎng)液,連續(xù)灌服一周：結(jié)果表明，日糧中單獨(dú)添加溶菌酶或60/120 mg/kg的EO可顯著降低肉仔雞的死亡率(P0.05),降低d 21肝臟細(xì)菌移位數(shù)(P0.05)。并且,單獨(dú)添加溶菌酶或EO可不同程度地減輕d 21的腸道損傷評(píng)分,改善小腸組織形態(tài)，提高腸黏膜二糖酶活力和血清溶菌酶活力。其中,日糧中單獨(dú)添加120 mg/kg的EO顯著提高了小腸iNOS、 occludin和claudin l的mRNA表達(dá)(P0.05)。 16s rRNA基因測(cè)序和宏基因組測(cè)序結(jié)果表明120 mg/kg EO的添加改變了回腸菌群結(jié)構(gòu)和微生物功能基因的組成,提高了梭菌目和棒狀桿菌屬細(xì)菌的相對(duì)含量(P0.05),抑制了某些致病菌的毒力因子基因的相對(duì)豐度(P0.05)。整體來(lái)看,日糧中單獨(dú)添加120 mg/kg EO對(duì)肉仔雞的改善效果最好,而本試驗(yàn)所用的溶菌酶與EO聯(lián)合使用不能增強(qiáng)其改善壞死性腸炎的作用效果。以上研究表明,日糧中添加以百里香酚和香芹酚為活性成分的EO具有顯著的免疫調(diào)節(jié)作用,能夠直接影響LPS誘導(dǎo)的肉仔雞炎癥反應(yīng),并且能夠影響腸道微生物區(qū)系,改善腸上皮屏障功能,緩解產(chǎn)氣莢膜梭菌誘導(dǎo)的肉仔雞壞死性腸炎。但是,EO不能有效緩解球蟲感染所造成的腸道損傷,本試驗(yàn)所用的外源溶菌酶與EO配伍使用不能增強(qiáng)EO對(duì)壞死性腸炎的改善作用。
[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.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S831.5
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本文編號(hào)：2005865