利用16S rDNA测序技术分析腹泻仔猪肠道菌群结构特征及灌服粪肠球菌对其调节作用

发布时间：2018-02-08 09:17

本文关键词： 腹泻粪肠球菌肠道微生物宏基因组学 16SrDNA　出处：《南京农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：肠道微生物与机体的健康息息相关。仔猪在哺乳期因生理缺陷、外界环境压力、食物变化等原因常导致肠道菌群紊乱,发生腹泻。腹泻同样会加剧肠道微生物的紊乱,生产性能下降,并造成巨大经济损失。有研究表明宿主的肠道微生物与性别相关,但也一些截然相反的结果。微生态制剂作为一种理想的抗生素替代品常用来调节猪肠道微生物平衡,预防和治疗仔猪腹泻。本文利用16SrDNA技术分析腹泻仔猪肠道菌群结构特征及灌服微生态制剂对仔猪肠道微生物的内在影响及其性别差异。本实验选择胎次、产期、产仔数相近的新生仔猪48窝,观察仔猪群健康状况,一旦发现某窝仔猪发生腹泻,立即选择该窝腹泻仔猪1头及同窝同性别未腹泻的仔猪1头采集血样、粪样,直至采集20窝仔猜样,并将这20窝仔猪随机分对照组(C)、试验组1(T1)、试验组2(T2)、试验组3(T3)、试验组4(T4)、每组4窝。五个组于腹泻当日(0d)、第3日(3d)、第6日(6d)全窝分别灌服双蒸水1mL(C)、粪肠球菌2.5 × 108 CFU/mL(T1)、5 × 108 CFU/mL(T2)、1 × 109 CFU/mL(T3)、2 × 109 CFU/mL(T4)的微生态制剂1mL。到20日龄时,每窝选择试验期间发生过腹泻与同性别从未腹泻的仔猪各一头采集血液样品和粪样。试验分为两个部分:1腹泻仔猪肠道微生物、免疫指标的情况本实验利用16SrDNA技术分析腹泻仔猪肠道微生物特征及其性别差异。结果表明:腹泻仔猪与未腹泻仔猪肠道微生物多样性没有差异,但在门水平上,腹泻组疣微菌门Verrucomicrobia(P=0.0419)的丰度显著高于未腹泻组;在属水平上,腹泻组消化链球菌属Peptostreptococcus(P=0.0195)的丰度显著高于未腹泻组,而丛毛单胞菌属Comamonas(P=0.0308)和Peptoniphilus(=0.0243)的丰度显著低于未腹泻组;公仔猪组中 Butyricimonas(P=0.0019)、Sharpea(P=0.0049)和Akkermansia(P=0.0168)的丰度显著显著高于母仔猪组。腹泻仔猪血液中免疫球蛋白M(IgM)浓度显著低于未腹泻仔猪组(P=0.049)。2灌服粪肠球菌微生态制剂对仔猪肠道微生物、免疫指标的影响参考本实验室前期关于粪肠球菌微生态制剂对仔猪腹泻、血液指标及生长性能影响的研究结果,挑选对照组(C)、试验组2(T2)、试验组4(T4)中同单元、同性别仔猪各6头,用以研究灌服粪肠球菌微生态制剂对仔猪肠道微生物的影响。结果表明,灌服粪肠球菌微生态制剂有治疗腹泻的功效,但没有改变仔猪肠道微生物的多样性和丰度。与对照组相比,灌服微生态制剂显著提高血液中IgA(P=0.0250)、IgM(P=0.0402)的浓度。此外,通过两个时间段对照组肠道微生物的比较,我们发现仔猪肠道微生物的多样性和丰度随日龄改变;在门水平,3-7日龄组梭杆菌门Fusobacteria(P=0.015)的丰度显著低于20日龄组,但螺旋体门Spirochaetes(P=0.045)和纤维杆菌门Fibrobacteres(P=0.043)的丰度显著高于20日龄组。
[Abstract]:Intestinal microbes are closely related to the health of the body. During lactation, piglets often suffer from intestinal microflora disorders due to physiological defects, environmental pressures, food changes, and other reasons. Diarrhea also exacerbates intestinal microbiological disorders. Poor productivity and huge economic losses. Studies have shown that intestinal microbes in the host are gender-related. As an ideal substitute for antibiotics, microecological agents are often used to regulate the microbial balance in pig intestines. To prevent and treat diarrhea in piglets, 16s rDNA technique was used to analyze the characteristics of intestinal microflora in piglets, the internal effects of microecological preparations on intestinal microbes and their sex differences. 48 litters of newborn piglets with similar litter size were observed. Once diarrhea was found in one litter piglet, one piglet with diarrhea and one piglet of the same litter with no diarrhea of the same sex were selected to collect blood samples and feces. Until the collection of 20 litter guess, The 20 piglets were randomly divided into two groups: control group (n = 1), test group (n = 1), test group (n = 2), test group (n = 3), test group (n = 3), test group (n = 4), group B (n = 4), each group (n = 4). The five groups were given double distilled water (1 mL C) and enterococcus faecalis (2.5 脳 10 ~ 8 CFU / m L ~ (1) T _ (1)) on the day of diarrhea. A microecological preparation of 1 脳 10 9 CFU / mL T3, 2 脳 10 9 CFU / mL T4). By the age of 20 days, Blood samples and feces were collected from one piglet with diarrhea during each litter selection trial and one piglet with no diarrhea of the same sex. The experiment was divided into two parts: 1. The characteristics of intestinal microbes and their sex differences in diarrhea piglets were analyzed by 16s rDNA technique. The results showed that there was no difference in intestinal microbial diversity between diarrhea piglets and non-diarrhea piglets, but at the portal level, there was no significant difference in intestinal microbial diversity between diarrhea piglets and non-diarrhea piglets. The abundance of Verrucomicrobiae (0.0419) in diarrhea group was significantly higher than that in non-diarrhea group, and the abundance of Peptostreptococcus P0. 0195 in diarrhea group was significantly higher than that in non diarrhea group, while the abundance of Comamonas P0.0308) and Peptoniphilus 0.0243) in diarrhea group was significantly lower than that in non diarrhea group. The abundance of Butyric imonasone P0. 0019 Sharpea P0. 0049) and Akkermania P0. 0168) in male piglets was significantly higher than that in female piglets, and the concentration of immunoglobulin (IgM) in blood of diarrhea piglets was significantly lower than that in non diarrhea piglets (P0. 049.2), and the microecological preparation of Enterococcus faecalis on intestinal microorganism of piglets was significantly lower than that of non diarrhea piglets (P 0. 049. 2). The effect of immune index on the effect of enterococcus faecalis microecological preparation on diarrhea, blood index and growth performance of piglets was studied in our laboratory. The control group was selected as the control group, the control group was divided into two groups: the control group (n = 2), the test group (n = 4) and the control group (n = 6). To study the effect of enterococcus faecalis microecological preparation on intestinal microorganism of piglets. The results showed that enterococcus faecalis microecological preparation had the effect of treating diarrhea. But it did not change the diversity and abundance of intestinal microorganism in piglets. Compared with the control group, the concentration of IgAP0. 0250 in blood was significantly increased by using microecological preparation. In addition, compared with the control group, the intestinal microbes in the control group were compared in two time periods. We found that the diversity and abundance of intestinal microorganism in piglets varied with age, and the abundance of Fusobacteriae (P0. 015) was significantly lower than that of 20 days old group, but the abundance of spirochaetes P0. 045) and Fibrobacteres P0. 043) were significantly higher than those of 20 days old group.
【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.28

【参考文献】