以16S rDNA高通量测序法研究果寡糖对奶牛瘤胃菌群结构及多样性的影响

doi:10.16431/j.cnki.1671-7236.2018.10.013

摘要/Abstract

摘要：

试验通过16S rDNA高通量测序技术研究果寡糖对奶牛瘤胃菌群结构及多样性的影响。采用2×2交叉设计，选用4头泌乳阶段相近、胎次相同的中国荷斯坦泌乳奶牛为试验动物，随机分为2组（试验组和对照组），对照组饲喂基础日粮，试验组在基础日粮中添加果寡糖60 g/（d·头），每期21 d，预饲期14 d，正试期7 d，交叉试验过渡期14 d。分别于饲喂前（0 h）和饲喂后2、4、6、9、12 h通过口腔导管采集瘤胃液，每期连续采集3 d（第16、17、18天），利用16S rDNA高通量测序技术测定瘤胃液中菌群结构及其多样性。结果显示，从Alpha多样性指数（Simpson、Shannon）、丰富度指数（Chao、Ace）及Beta diversity热图可知，果寡糖具有降低奶牛瘤胃中细菌多样性的趋势。从门水平上分析，试验组及对照组中奶牛瘤胃中拟杆菌门、厚壁菌门和变形细菌门占总细菌的95%以上，果寡糖显著降低了奶牛瘤胃未注释细菌门的丰度（P<0.05），互养菌门丰度有增加的趋势（P=0.075）。从属水平上分析，果寡糖显著提高了奶牛瘤胃假单胞菌属丰度（P<0.05），极显著提高了拟杆菌属丰度（P<0.01），而脱盐杆菌属丰度显著降低（P<0.05）。与对照组相比，添加果寡糖的试验组奶牛瘤胃中瘤胃球菌属、丁酸弧菌属、琥珀酸菌属、未注释毛螺菌属丰度分别增加80.0%、7.5%、127.9%和20.0%，但差异均不显著（P>0.05）。综上所述，本试验条件下，果寡糖的添加对奶牛瘤胃细菌菌群多样性及结构存在一定的影响，果寡糖显著提高了奶牛瘤胃中不发酵糖类的假单胞菌属的丰度，使碳水化合物为发酵源的瘤胃拟杆菌丰度极显著增加，对瘤胃纤维降解菌丰度的增加具有促进作用。

关键词: 果寡糖; 中国荷斯坦奶牛; 瘤胃细菌菌群; 16S rDNA高通量测序技术

Abstract:

This experiment was aimed to study the effect of fructooligosaccharides on the structure and diversity of rumen bacterial community in dairy cows by 16S rDNA sequencing technology.Four lactating dairy cows with similar lactation stages and the same parity were randomly divided into two groups using two-phase cross design method.The cows in control group were fed with basal diet and that in test group were fed with the basal diet with 60 g/(d·head) fructooligosaccharides.The experiment was designed through 2×2 cross-over test with 21 d in each stage (14 d for pre-experiment and 7 d for test) and 14 d transitional period of cross test.The rumen fluid samples were collected at 0 (before feeding),2,4,6,9 and 12 h after feeding by cattle catheter.Each phase was collected continuously for 3 days,and the structure and diversity of rumen bacterial community were measured by 16S rDNA sequencing technology.The results of Alpha diversity indexes (Simpson,Shannon),richness indexes (Chao,Ace) and Beta diversity profile showed that the addition of fruitooligosaccharide decreased the bacterial diversity in the rumen of dairy cows.And the results of 16S rDNA sequencing showed that according to the analysis at phylum level,Bacteroidetes,Firmicutes and Proteobacteria accounted for more than 95% of the total bacterial population in cow rumen of two groups.The addition of fructooligosaccharides significantly decreased the abundance of Bacteria_NA (P < 0.05),and the abundance of Synergistetes showed an increasing trend (P=0.075).According to the analysis at genus level,fructooligosaccharides significantly increased the abundance of the Pseudomonas (P < 0.05),extremely significantly increased the abundance of Bacteroides (P < 0.01),while the abundance of Dehalobacterium significantly decreased (P < 0.05).Compared with the control group,the abundance of Ruminococcus,Butyrivibrio,Succiniclasticum and Lachnospiraceae_NA in test group was increased by 80.0%,7.5%,127.9% and 20.0%,respectively,but these differences were not significant (P > 0.05).Under the test conditions,the addition of fructooligosaccharides had a certain effect on the diversity and structure of rumen bacterial flora in dairy cows,which significantly increased the abundance of Pseudomonas which unfermented sugar,extremely significantly increased the abundance of Bacteroides which used carbohydrates as a source of fermentation,had an accelerating effect on the abundance of rumen fiber degrading bacteria.

Key words: fructooligosaccharides; Chinese Holstein dairy cows; rumen bacterial flora; 16S rDNA sequencing technology

中图分类号:

S816.7

胡丹丹, 郭婷婷, 李娜, 张力莉, 徐晓锋. 以16S rDNA高通量测序法研究果寡糖对奶牛瘤胃菌群结构及多样性的影响[J]. 《中国畜牧兽医》, 2018, 45(10): 2761-2771.

HU Dandan, GUO Tingting, LI Na, ZHANG Lili, XU Xiaofeng. Effect of Fructooligosaccharide on the Structure and Diversity of Rumen Bacterial Community in Dairy Cow by 16S rDNA Sequencing Technology[J]. , 2018, 45(10): 2761-2771.

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