绞股蓝内生真菌JY25胞外多糖对肉鸡生长性能和肠黏膜免疫功能的影响

发布时间：2018-07-24 09:39

【摘要】：药用植物内生真菌是一种新型的药用微生物资源,其产生的真菌胞外多糖是一种新型绿色饲料添加剂,也具有真菌多糖的多种生物学特性,如高效、无毒、无残留,能提高机体免疫力、抗病能力、提高畜禽的生产性能等。本研究以药用植物绞股蓝内生真菌JY25发酵液中提取的的胞外多糖(以下简称JY25P)为实验材料,通过体内外试验,观察JY25P对肉鸡生长、免疫特性的影响,并通过对体外培养T、B淋巴细胞和肠黏膜免疫细胞分泌细胞因子的影响,探索药用植物内生真菌多糖增强肉鸡肠道免疫性能的机制,为其实际生产应用提供理论依据和基础数据。试验一JY25P对肉鸡生长性能和肠道结构的影响在肉鸡日粮中全程添加不同浓度的JY25P,通过测定肉鸡平均日增重(ADG)、平均日采食量(ADFI)、料重比(FGR)研究其对肉鸡生长性能的影响;检测不同试验组肉鸡十二指肠和空肠的绒毛高度、隐窝深度、绒毛高度/隐窝深度,以分析促进肠道营养物质吸收的机制。结果显示,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分别降低了12.63%(P0.05)和20.50%(P0.05),证实日粮中添加JY25P可以提高肉鸡生长性能,从而提高经济效益;从饲喂JY25P的肠道结构分析发现:添加0.4%的JY25P在42日龄可以提高肉鸡十二指肠绒毛高度和V/C值分别提高了19.73%(P0.05)和24.08%(P0.05),空肠绒毛高度和V/C值分别提高了16.03%(P0.05)和25.35%(P0.05),证实JY25P可以改善肉鸡小肠肠道结构,从而可以增强机体消化吸收功能。综合考虑JY25P对肉鸡生长性能和肠道结构的影响,日粮中JY25P的添加量为0.4%时生产效益最好。试验二JY25P提高肉鸡免疫性能的机制研究通过检测全程饲喂JY25P肉鸡的新城疫抗体效价和肠道黏膜SIgA、IL-4、IFN-γ免疫因子的变化以分析JY25P提高肉鸡免疫能力的机制。结果显示,添加0.6%的JY25P,在21、28、35、42日龄分别提高新城疫抗体效价35.77%、35.14%、25.00%、33.50%,差异显著(P0.05),增强了机体的特异性免疫能力;JY25P能够增加肠道黏膜分泌SIgA、IL-4和IFN-γ(P0.05),提高机体体液免疫和细胞免疫水平,同时维持肠道黏膜免疫平衡,促进机体的非特异性免疫。由此可见,JY25P的添加可增强特异性免疫能力、提高肠黏膜非特异性免疫性能达到增强机体免疫、抗病的作用。试验三JY25P对体外培养淋巴细胞增殖和肠黏膜淋巴细胞因子的影响采用MTT法检测JY25P对体外培养肉鸡外周血及不同免疫器官的T、B淋巴细胞增殖的影响;以不同剂量的JY25P刺激肉鸡肠道派伊尔淋巴细胞(PPL)、肠上皮内淋巴细胞(IEL)、固有层淋巴细胞(LPL)分泌IL-2、IL-10、TNF-α免疫因子。试验结果显示,添加80μg/mL的JY25P可刺激体外培养的外周血、胸腺、脾脏T淋巴细胞的增殖,分别提高142.23%(P0.01)、42.05%(P0.05)、35.09%(P0.01),添加80μg/mL的JY25P在体外直接刺激外周血、脾脏、法氏囊B淋巴细胞的增殖,分别提高67.99%(P0.01)、68.47%(P0.01)、48.73%(P0.05),说明JY25P可以增强淋巴细胞的增殖;添加160μg/mL的JY25P,可显著提高淋巴细胞抗炎细胞因子(IL-10)的表达(P0.05),抑制炎性细胞因子(IL-2、TNF-α)的表达(P0.05),在一定程度上调节黏膜免疫状态、促进组织修复,达到治疗和缓解黏膜免疫紊乱的作用。综上所述,日粮中添加0.4%~0.6%的JY25P可以促进肉鸡的生长,通过改善肠道绒毛和隐窝结构促进营养物质的消化吸收,提高生产效益;同时,提高血清中新城疫抗体效价、促进肠黏膜分泌免疫因子,增强机体特异性和非特异性免疫能力;通过体外试验证实,一定浓度的JY25P能够刺激肉鸡T、B淋巴细胞的增殖,调整肠黏膜免疫细胞分泌淋巴细胞因子,从而调节肠黏膜的免疫状态,增强肉鸡肠道免疫和抗病能力,从而达到综合提高生产性能的效果。
[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.
【学位授予单位】：河南工业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S831.5

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