【摘要】：藥用植物內(nèi)生真菌是一種新型的藥用微生物資源,其產(chǎn)生的真菌胞外多糖是一種新型綠色飼料添加劑,也具有真菌多糖的多種生物學(xué)特性,如高效、無毒、無殘留,能提高機(jī)體免疫力、抗病能力、提高畜禽的生產(chǎn)性能等。本研究以藥用植物絞股藍(lán)內(nèi)生真菌JY25發(fā)酵液中提取的的胞外多糖(以下簡(jiǎn)稱JY25P)為實(shí)驗(yàn)材料,通過體內(nèi)外試驗(yàn),觀察JY25P對(duì)肉雞生長(zhǎng)、免疫特性的影響,并通過對(duì)體外培養(yǎng)T、B淋巴細(xì)胞和腸黏膜免疫細(xì)胞分泌細(xì)胞因子的影響,探索藥用植物內(nèi)生真菌多糖增強(qiáng)肉雞腸道免疫性能的機(jī)制,為其實(shí)際生產(chǎn)應(yīng)用提供理論依據(jù)和基礎(chǔ)數(shù)據(jù)。試驗(yàn)一JY25P對(duì)肉雞生長(zhǎng)性能和腸道結(jié)構(gòu)的影響在肉雞日糧中全程添加不同濃度的JY25P,通過測(cè)定肉雞平均日增重(ADG)、平均日采食量(ADFI)、料重比(FGR)研究其對(duì)肉雞生長(zhǎng)性能的影響;檢測(cè)不同試驗(yàn)組肉雞十二指腸和空腸的絨毛高度、隱窩深度、絨毛高度/隱窩深度,以分析促進(jìn)腸道營(yíng)養(yǎng)物質(zhì)吸收的機(jī)制。結(jié)果顯示,添加0.4%的JY25P平均末重提高了6.40%(P0.05),ADG提高了9.89%(P0.05),ADFI和FGR分別降低了12.63%(P0.05)和20.50%(P0.05),證實(shí)日糧中添加JY25P可以提高肉雞生長(zhǎng)性能,從而提高經(jīng)濟(jì)效益;從飼喂JY25P的腸道結(jié)構(gòu)分析發(fā)現(xiàn):添加0.4%的JY25P在42日齡可以提高肉雞十二指腸絨毛高度和V/C值分別提高了19.73%(P0.05)和24.08%(P0.05),空腸絨毛高度和V/C值分別提高了16.03%(P0.05)和25.35%(P0.05),證實(shí)JY25P可以改善肉雞小腸腸道結(jié)構(gòu),從而可以增強(qiáng)機(jī)體消化吸收功能。綜合考慮JY25P對(duì)肉雞生長(zhǎng)性能和腸道結(jié)構(gòu)的影響,日糧中JY25P的添加量為0.4%時(shí)生產(chǎn)效益最好。試驗(yàn)二JY25P提高肉雞免疫性能的機(jī)制研究通過檢測(cè)全程飼喂JY25P肉雞的新城疫抗體效價(jià)和腸道黏膜SIgA、IL-4、IFN-γ免疫因子的變化以分析JY25P提高肉雞免疫能力的機(jī)制。結(jié)果顯示,添加0.6%的JY25P,在21、28、35、42日齡分別提高新城疫抗體效價(jià)35.77%、35.14%、25.00%、33.50%,差異顯著(P0.05),增強(qiáng)了機(jī)體的特異性免疫能力;JY25P能夠增加腸道黏膜分泌SIgA、IL-4和IFN-γ(P0.05),提高機(jī)體體液免疫和細(xì)胞免疫水平,同時(shí)維持腸道黏膜免疫平衡,促進(jìn)機(jī)體的非特異性免疫。由此可見,JY25P的添加可增強(qiáng)特異性免疫能力、提高腸黏膜非特異性免疫性能達(dá)到增強(qiáng)機(jī)體免疫、抗病的作用。試驗(yàn)三JY25P對(duì)體外培養(yǎng)淋巴細(xì)胞增殖和腸黏膜淋巴細(xì)胞因子的影響采用MTT法檢測(cè)JY25P對(duì)體外培養(yǎng)肉雞外周血及不同免疫器官的T、B淋巴細(xì)胞增殖的影響;以不同劑量的JY25P刺激肉雞腸道派伊爾淋巴細(xì)胞(PPL)、腸上皮內(nèi)淋巴細(xì)胞(IEL)、固有層淋巴細(xì)胞(LPL)分泌IL-2、IL-10、TNF-α免疫因子。試驗(yàn)結(jié)果顯示,添加80μg/mL的JY25P可刺激體外培養(yǎng)的外周血、胸腺、脾臟T淋巴細(xì)胞的增殖,分別提高142.23%(P0.01)、42.05%(P0.05)、35.09%(P0.01),添加80μg/mL的JY25P在體外直接刺激外周血、脾臟、法氏囊B淋巴細(xì)胞的增殖,分別提高67.99%(P0.01)、68.47%(P0.01)、48.73%(P0.05),說明JY25P可以增強(qiáng)淋巴細(xì)胞的增殖;添加160μg/mL的JY25P,可顯著提高淋巴細(xì)胞抗炎細(xì)胞因子(IL-10)的表達(dá)(P0.05),抑制炎性細(xì)胞因子(IL-2、TNF-α)的表達(dá)(P0.05),在一定程度上調(diào)節(jié)黏膜免疫狀態(tài)、促進(jìn)組織修復(fù),達(dá)到治療和緩解黏膜免疫紊亂的作用。綜上所述,日糧中添加0.4%~0.6%的JY25P可以促進(jìn)肉雞的生長(zhǎng),通過改善腸道絨毛和隱窩結(jié)構(gòu)促進(jìn)營(yíng)養(yǎng)物質(zhì)的消化吸收,提高生產(chǎn)效益;同時(shí),提高血清中新城疫抗體效價(jià)、促進(jìn)腸黏膜分泌免疫因子,增強(qiáng)機(jī)體特異性和非特異性免疫能力;通過體外試驗(yàn)證實(shí),一定濃度的JY25P能夠刺激肉雞T、B淋巴細(xì)胞的增殖,調(diào)整腸黏膜免疫細(xì)胞分泌淋巴細(xì)胞因子,從而調(diào)節(jié)腸黏膜的免疫狀態(tài),增強(qiáng)肉雞腸道免疫和抗病能力,從而達(dá)到綜合提高生產(chǎn)性能的效果。
[Abstract]:Endophytic fungi of medicinal plants are a new kind of medicinal microbial resources. The extracellular polysaccharide produced by fungi is a new kind of green feed additive, and it also has many biological characteristics of fungal polysaccharides, such as high efficiency, non-toxic and no residue. It can improve the immunity of the body, resistance to disease, and improve the production performance of livestock and poultry. This study is a medicinal plant. Extracellular polysaccharide (hereinafter referred to as JY25P) extracted from the Gynostemma pentaphyllum endophyte JY25 fermentation broth was used as the experimental material. The effects of JY25P on the growth and immune characteristics of broilers were observed in vitro and in vitro. The effects of T, B lymphocytes and intestinal mucosal immune cells on the secretion of fine cell factors in vitro were investigated to explore the increase of Polysaccharides from endophytic fungi in medicinal plants. The mechanism of intestinal immune performance of strong broilers provides theoretical basis and basic data for its practical application. The effects of JY25P on growth performance and intestinal structure of broilers are added with different concentrations of JY25P in broiler diet, by measuring average daily weight gain (ADG), average daily feed intake (ADFI) and feed weight ratio (FGR) for meat in broilers. The effects of chicken growth performance; detection of the villus height of duodenum and jejunum in different test groups, the depth of the recess, the height of the villi / recess depth to analyze the mechanism of promoting the absorption of intestinal nutrients. The results showed that the average end weight of 0.4% JY25P increased by 6.40% (P0.05), ADG increased by 9.89% (P0.05), and ADFI and FGR decreased by 12.63% respectively. P0.05) and 20.50% (P0.05), confirmed that adding JY25P in the diet could improve the growth performance of Broilers and improve economic benefits. From the intestinal structure analysis of feeding JY25P, it was found that adding 0.4% JY25P at 42 days of age could increase the height of duodenum villus and V/C value of broilers to increase 19.73% (P0.05) and 24.08% (P0.05), and the height and V/C of jejunum villus. 16.03% (P0.05) and 25.35% (P0.05) were improved respectively. It was proved that JY25P could improve the intestinal intestinal structure of Broilers and enhance the body's digestive and absorption function. The effect of JY25P on growth performance and intestinal structure of broilers was considered. The production benefit was best when the dosage of JY25P was 0.4% in the diet. Test two JY25P to improve the immune performance of broilers. By detecting the antibody titer of Newcastle disease in JY25P broilers and the changes of SIgA, IL-4, and IFN- gamma immune factors in the intestinal mucosa to analyze the mechanism of JY25P to improve the immune ability of broilers, the results showed that the antibody titer of Newcastle disease was increased by 35.77%, 35.14%, 25%, 33.50% at 21,28,35,42 days (P0.05), respectively (P0.05). It can enhance the specific immunity of the body; JY25P can increase the secretion of SIgA, IL-4 and IFN- gamma (P0.05) in the intestinal mucosa, improve the body humoral immunity and cellular immunity level, and maintain the intestinal mucosal immune balance and promote the non specific immunity of the body. Thus, the addition of JY25P can enhance the specific immunity and improve the intestinal mucosa nonspecific. The effect of heterosexual immunity can enhance the immune and disease resistance. Experiment three JY25P on the proliferation of lymphocyte and the influence of intestinal mucosal lymphocyte factor in vitro, the effect of JY25P on the proliferation of T and B lymphocyte in peripheral blood and different immune organs of broilers in vitro was detected by MTT, and the intestinal pies of broiler were stimulated with different doses of JY25P. IL lymphocyte (PPL), intestinal intraepithelial lymphocyte (IEL) and lamina propria lymphocyte (LPL) secrete IL-2, IL-10, TNF- alpha immune factors. Experimental results showed that the addition of 80 mu JY25P could stimulate the proliferation of peripheral blood, thymus and spleen T lymphocytes in vitro, increased by 142.23% (P0.01), 42.05% (P0.05), 35.09% (P0.01), and 80 micron. The proliferation of peripheral blood, spleen, and bursal B lymphocyte proliferation by JY25P in vitro increased by 67.99% (P0.01), 68.47% (P0.01), 48.73% (P0.05), indicating that JY25P could enhance the proliferation of lymphocyte, and the expression of IL-10 (P0.05) was significantly increased by the addition of 160 mu JY25P (P0.05), and inflammatory cytokines were inhibited (IL-2, TN). The expression of F- alpha (P0.05), to some extent, regulates the mucosal immune state, promotes tissue repair, and achieves the effect of treating and alleviating the mucosal immune disorder. To sum up, the addition of 0.4%~0.6% JY25P in the diet can promote the growth of broiler, and improve the digestion and absorption of nutrients by improving the intestinal villi and recess structure, and improve the production efficiency. At the same time, the antibody titer of Newcastle disease in the serum was enhanced, the intestinal mucosa secreted the immune factors and the body specific and non specific immune ability was enhanced. Through in vitro experiments, it was proved that a certain concentration of JY25P could stimulate the proliferation of T and B lymphocytes in Broilers and adjust the intestinal mucosal immune cell secreting lymphocyte factors, thus regulating the immunity of intestinal mucosa. The state enhances the intestinal immunity and disease resistance of the broiler, so as to achieve the comprehensive improvement of the production performance.
【學(xué)位授予單位】：河南工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S831.5

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