益生菌对蛋鸡生产性能及肠道微生物变化与基因表达研究

发布时间：2018-04-26 14:11

本文选题：蛋鸡 + 益生菌　；参考：《四川农业大学》2016年硕士论文

【摘要】：动物的胃肠道内存在着大量微生物,可以影响宿主的消化、代谢、营养、免疫等各个方面。过去的研究中关于益生菌对肠道微生物菌群和宿主免疫影响的研究比较少,而高通量测序技术的进步,提供了一个更深入的了解细菌多样性和微生物菌群与动物相互作用的途径。目前,关于优化鸡肠道微生物菌群的干预措施已经成为研究的重点。本试验选用600羽罗曼粉壳蛋鸡,通过添加不同益生菌研究对生产性能的影响,并利用荧光定量PCR技术检测肝脏中生长激素(GH)、生长激素受体(GHR)、胰岛素样生长因子1 (IGF-1)基因的相对表达量,采集的盲肠组织通过扩增总16S rDNA的V4-V5区来分析样品的微生物组成。采集卵巢组织,利用荧光定量PCR技术检测雌激素受体-α受体(ERa)、雌激素受体-p受体(ERβ)、极低密度脂蛋白受体(VLDLR)、催乳素受体(PRLR)基因的相对表达量。主要试验结果如下：添加戊糖片球菌可以在3-9周龄显著提高蛋鸡体重(P0.05),可以增重4%左右,而添加枯草芽孢杆菌的个体则体重最轻,添加植物乳杆菌对影响蛋鸡体重方面效果不明显。但是益生菌对蛋鸡的体重影响在后期逐渐变小,各组之间的体重差异不再显著(P0.05)。添加添加枯草芽孢杆菌的个体在产蛋高峰期产蛋率显著高于其他组(P0.05),最高可达97.98%,并且可以延长蛋鸡产蛋高峰,相对于对照组多了两周。而添加植物乳杆菌的个体产蛋率是四组之间最低,添加戊糖片球菌则对个体的产蛋率影响不明显。添加益生菌并没有对初产体重、初产蛋重和初产日龄产生显著影响(P0.05)。添加枯草芽孢杆菌可以极显著提高蛋黄重、蛋重和长径(P0.01),分别比对照组高4.16%、3.22%和2.23%,并且显著提高了蛋壳平均厚度和短径(P0.05),添加植物乳杆菌可以极显著提高蛋黄颜色(P0.01),而添加戊糖片球菌对蛋品质未产生明显效果。通过16S rDNA测序共得到285604个有效序列,每个样品平均有效序列为23800+2338条,共得到347个OTUs。肠道微生物主要由由拟杆菌门、厚壁菌门和变形菌门3个菌门组成,占了96%以上,但是这3个菌门在各组中所占的百分比有较大差异,拟杆菌门比率最高的为LP组的79.50%,最低的为CP组的63.50%；厚壁菌门比率最高的为CP组的29.20%,最低为PP组的14.50%;变形菌门比率最高的为PP组的4.90%,最低的为SB组的2.30%。四组样品的肠道微生物中的Oxalobacter和S24-7_norank这两个菌属差异显著(P0.05),并且S24-7_norank这个菌属只在添加戊糖片球菌的个体中存在。各个处理组分别与对照组进行比较,发现添加戊糖片球菌的个体共有6种菌属有显著的倍数变化(P0.05),分别为Oscillibacter,Oxalobacter, Bacteroides, Barnesiella, Gastranaerophilales, S24-7_norank菌属,添加枯草芽孢杆菌的个体有5种菌属有显著的倍数变化(P0.05),分别为Bacteroides, Barnesiella, Fusobacterium, Gastranaerophilales, Rikenella,而添加植物乳杆菌的个体只有Odoribacter菌属有显著的倍数变化(P0.05)通过荧光定量PCR分析发现添加戊糖片球菌的个体肝脏中GH和GHR表达量显著增加(P0.05),添加枯草芽孢杆菌的个体卵巢中PRLR的表达量和肝脏中GHR的表达量显著下降(P0.05)。在产蛋期长期添加枯草芽孢杆菌的个体卵巢中ERβ, ERα基因的表达量显著增加(P0.05),并且显著降低了跟产蛋率负相关的PRLR表达(P0.05)。上述结果说明早期定植益生菌可以对蛋鸡肠道微生物组成产生影响,进而促进或抑制蛋鸡的生长发育过程,并且这种影响有可能是通过调节宿主自身基因表达造成的,然而这种影响具有分阶段性,不同益生菌产生的影响也不相同。
[Abstract]:There are a large number of microbes in the gastrointestinal tract of the animal, which can affect the digestive, metabolic, nutritional and immune aspects of the host. In the past, there are few studies on the effects of probiotics on the intestinal microflora and host immunity, and the progress of high flux sequencing technology provides a more thorough understanding of bacterial diversity and microbiology. At present, the intervention measures for optimizing the microbial flora of Chicken Intestines have been the focus of the study. 600 Roman shell layers were selected in this experiment. The effects of different probiotics on the production performance were added, and the growth hormone (GH) and growth hormone in the liver were detected by the fluorescence quantitative PCR technique. The relative expression of the receptor (GHR), the insulin like growth factor 1 (IGF-1) gene, and the collected cecum tissue by amplification of the V4-V5 region of the total 16S rDNA to analyze the microorganism composition of the samples. The ovarian tissue was collected and the estrogen receptor alpha receptor (ERa), the estrogen receptor -p receptor (ER beta), and the extremely low density lipoprotein receptor (LDLR) were detected by the fluorescence quantitative PCR technique. VLDLR) the relative expression of the prolactin receptor (PRLR) gene. The main results are as follows: adding pentose Micrococcus can significantly increase the body weight (P0.05) at the age of 3-9 weeks and increase the weight by about 4%, while the individual with Bacillus subtilis is the least body weight, but the effect of adding Lactobacillus plantarum on the weight of laying hens is not obvious. The effect of bacteria on the weight of laying hens gradually became smaller in the later period, and the weight difference between each group was no longer significant (P0.05). The egg production rate of the addition of Bacillus subtilis was significantly higher than that of the other groups (P0.05), up to 97.98%, and the egg production peak could be extended to the control group for two weeks, and the addition of plant milk was added. The individual egg production rate of bacilli was the lowest among the four groups, and the addition of pentose Micrococcus had no significant influence on the egg production rate of the individual. The addition of probiotics did not have a significant effect on the initial birth weight, the egg weight and the day of birth (P0.05). The addition of Bacillus subtilis could significantly increase the yolk weight, the egg weight and the length (P0.01), higher than the control group, respectively. 4.16%, 3.22% and 2.23%, and significantly increased the average thickness and short diameter of eggshell (P0.05). Adding Lactobacillus plantarum could significantly improve the yolk color (P0.01), while adding pentose pentose coccus had no obvious effect on egg quality. 285604 effective sequences were obtained by 16S rDNA sequencing. The average effective sequence of each sample was 23800+2338. 347 OTUs. intestinal microbes were mainly composed of 3 bacteriobacteria, pthickens and deformable bacteria doors, accounting for more than 96%, but the percentage of the 3 bacteria doors in each group was different. The highest percentage of the bacteriobacteria was 79.50% in the LP group, the lowest in the CP group, and the highest in the CP group 29.20. The lowest was 14.50% of group PP, and the highest rate of deformable bacteria was 4.90% in group PP, and the lowest was Oxalobacter and S24-7_norank in the intestinal microbes of group 2.30%. four of group SB, which were significantly different (P0.05), and S24-7_norank was only in individuals adding pentose pentose. A total of 6 species of bacteria with pentose pentacose were found to have significant multiplier changes (P0.05), which were Oscillibacter, Oxalobacter, Bacteroides, Barnesiella, Gastranaerophilales, S24-7_norank, and 5 species with Bacillus subtilis, respectively, with a significant multiplier variation (P0.05), respectively, Bacteroide, respectively. S, Barnesiella, Fusobacterium, Gastranaerophilales, Rikenella, and the individuals adding Lactobacillus plantarum only had a significant multiplier variation (P0.05) by the fluorescence quantitative PCR analysis. The expression of GH and GHR in the individual liver added to pentacose streptococcus was significantly increased (P0.05), and the individual ovary of Bacillus subtilis was added. The expression of PRLR and the expression of GHR in the liver decreased significantly (P0.05). The expression of ER beta in the ovary of Bacillus subtilis was significantly increased (P0.05) at the laying period of Bacillus subtilis (P0.05), and the expression of PRLR (P0.05) negatively related to the egg production rate (P0.05) was significantly reduced. The results indicated that early colonization of probiotics could be used in the intestinal tract. The effect of the microbial composition on the growth and development of the laying hens may be caused by the regulation of the gene expression of the host's own, but the effect is phased, and the effects of different probiotics are different.

【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S831.5

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