本文選題：蛋雞 + 益生菌　； 參考：《四川農業(yè)大學》2016年碩士論文

【摘要】：動物的胃腸道內存在著大量微生物,可以影響宿主的消化、代謝、營養(yǎng)、免疫等各個方面。過去的研究中關于益生菌對腸道微生物菌群和宿主免疫影響的研究比較少,而高通量測序技術的進步,提供了一個更深入的了解細菌多樣性和微生物菌群與動物相互作用的途徑。目前,關于優(yōu)化雞腸道微生物菌群的干預措施已經成為研究的重點。本試驗選用600羽羅曼粉殼蛋雞,通過添加不同益生菌研究對生產性能的影響,并利用熒光定量PCR技術檢測肝臟中生長激素(GH)、生長激素受體(GHR)、胰島素樣生長因子1 (IGF-1)基因的相對表達量,采集的盲腸組織通過擴增總16S rDNA的V4-V5區(qū)來分析樣品的微生物組成。采集卵巢組織,利用熒光定量PCR技術檢測雌激素受體-α受體(ERa)、雌激素受體-p受體(ERβ)、極低密度脂蛋白受體(VLDLR)、催乳素受體(PRLR)基因的相對表達量。主要試驗結果如下：添加戊糖片球菌可以在3-9周齡顯著提高蛋雞體重(P0.05),可以增重4%左右,而添加枯草芽孢桿菌的個體則體重最輕,添加植物乳桿菌對影響蛋雞體重方面效果不明顯。但是益生菌對蛋雞的體重影響在后期逐漸變小,各組之間的體重差異不再顯著(P0.05)。添加添加枯草芽孢桿菌的個體在產蛋高峰期產蛋率顯著高于其他組(P0.05),最高可達97.98%,并且可以延長蛋雞產蛋高峰,相對于對照組多了兩周。而添加植物乳桿菌的個體產蛋率是四組之間最低,添加戊糖片球菌則對個體的產蛋率影響不明顯。添加益生菌并沒有對初產體重、初產蛋重和初產日齡產生顯著影響(P0.05)。添加枯草芽孢桿菌可以極顯著提高蛋黃重、蛋重和長徑(P0.01),分別比對照組高4.16%、3.22%和2.23%,并且顯著提高了蛋殼平均厚度和短徑(P0.05),添加植物乳桿菌可以極顯著提高蛋黃顏色(P0.01),而添加戊糖片球菌對蛋品質未產生明顯效果。通過16S rDNA測序共得到285604個有效序列,每個樣品平均有效序列為23800+2338條,共得到347個OTUs。腸道微生物主要由由擬桿菌門、厚壁菌門和變形菌門3個菌門組成,占了96%以上,但是這3個菌門在各組中所占的百分比有較大差異,擬桿菌門比率最高的為LP組的79.50%,最低的為CP組的63.50%；厚壁菌門比率最高的為CP組的29.20%,最低為PP組的14.50%;變形菌門比率最高的為PP組的4.90%,最低的為SB組的2.30%。四組樣品的腸道微生物中的Oxalobacter和S24-7_norank這兩個菌屬差異顯著(P0.05),并且S24-7_norank這個菌屬只在添加戊糖片球菌的個體中存在。各個處理組分別與對照組進行比較,發(fā)現(xiàn)添加戊糖片球菌的個體共有6種菌屬有顯著的倍數(shù)變化(P0.05),分別為Oscillibacter,Oxalobacter, Bacteroides, Barnesiella, Gastranaerophilales, S24-7_norank菌屬,添加枯草芽孢桿菌的個體有5種菌屬有顯著的倍數(shù)變化(P0.05),分別為Bacteroides, Barnesiella, Fusobacterium, Gastranaerophilales, Rikenella,而添加植物乳桿菌的個體只有Odoribacter菌屬有顯著的倍數(shù)變化(P0.05)通過熒光定量PCR分析發(fā)現(xiàn)添加戊糖片球菌的個體肝臟中GH和GHR表達量顯著增加(P0.05),添加枯草芽孢桿菌的個體卵巢中PRLR的表達量和肝臟中GHR的表達量顯著下降(P0.05)。在產蛋期長期添加枯草芽孢桿菌的個體卵巢中ERβ, ERα基因的表達量顯著增加(P0.05),并且顯著降低了跟產蛋率負相關的PRLR表達(P0.05)。上述結果說明早期定植益生菌可以對蛋雞腸道微生物組成產生影響,進而促進或抑制蛋雞的生長發(fā)育過程,并且這種影響有可能是通過調節(jié)宿主自身基因表達造成的,然而這種影響具有分階段性,不同益生菌產生的影響也不相同。
[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.

【學位授予單位】：四川農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S831.5

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