单宁与饲用纤维素酶对湖羊瘤胃微生物菌群的影响

doi:10.16431/j.cnki.1671-7236.2019.01.013

摘要/Abstract

摘要：

试验旨在利用高通量测序技术研究添加单宁与饲用纤维素酶对湖羊生长育肥期瘤胃微生物菌群结构的影响，为单宁和饲用纤维素酶在反刍动物生产中更好地利用提供理论依据。选用3月龄生长发育良好、平均体重（19.85±1.45） kg的肉用湖羊36只，随机分成对照组和3个处理组，对照组饲喂基础日粮，处理组分别在基础日粮中加入0.1%单宁（单宁组）、0.1%饲用纤维素酶（纤维素酶组）和0.1%单宁+0.1%饲用纤维素酶（混合组），每组3个重复，每个重复3只湖羊。试验期70 d，其中过渡期7 d，预试期7 d，正饲期56 d。试验结束后，采集湖羊瘤胃液，提取细菌总DNA，进行PCR扩增后用Illumina Hi Seq 2500测序平台进行高通量测序。结果表明：①12个湖羊瘤胃微生物样品测序共获得 957 440 对序列，平均每个样品产生55 997条clean tags，过滤嵌合体后共产生502 965条effective tags；各组湖羊瘤胃微生物样品的AvgLen均在419~420之间；②各组间湖羊瘤胃微生物的Ace指数和Chao1指数差异均不显著（P > 0.05）；与对照组相比，单宁组、纤维素酶组和混合组湖羊瘤胃微生物的香农指数显著提高（P < 0.05）；纤维素酶组湖羊瘤胃微生物的辛普森指数较对照组显著降低（P < 0.05）；③在门水平上，单宁组湖羊瘤胃内拟杆菌门（Bacteroidetes）丰度比对照组显著降低（P < 0.05）；单宁组、纤维素酶组和混合组湖羊瘤胃内厚壁菌门（Firmicutes）丰度较对照组均呈下降趋势，但差异不显著（P > 0.05）；④在属水平上，单宁组湖羊瘤胃内普雷沃菌属-1（Prevotella_1）丰度降低，与对照组、纤维素酶组差异均显著（P < 0.05），但与混合组差异不显著（P > 0.05）；单宁组和纤维素酶组湖羊瘤胃内瘤胃菌属（Rumen_bacterium）丰度较对照组和混合组提高，且与混合组的差异显著（P < 0.05）。综合试验结果，在湖羊日粮中同时加入单宁和饲用纤维素酶可以提高湖羊瘤胃内菌群的多样性和丰度，影响瘤胃菌群结构，缓解单宁对拟杆菌门（Bacteroidetes）和厚壁菌门（Firmicutes）菌落的抑制；同时混合添加可以缓解单独添加单宁对纤维素分解的抑制作用；在门水平上各组湖羊瘤胃内优势菌群均为拟杆菌门和厚壁菌门；在属水平上各组湖羊瘤胃内优势菌群均为理研菌科-RC9（Rikenellaceae_RC9_gut_group）、细菌（Bacterium）、普雷沃菌属-1和瘤胃菌属。

关键词: 单宁; 饲用纤维素酶; 湖羊; 微生物菌群; 高通量测序

Abstract:

The purpose of this study was to investigate the effects of adding tannins and feeding cellulase on rumen microbes in the growth and fattening period of Hu sheep using high-throughput sequencing technology,and provide useful data for the better utilization of tannins and cellulase in ruminant production.Thirty-six meat Hu sheep with good growth and average body weight (19.85±1.45) kg at 3 months old were randomly divided into control group and 3 treatment groups.The control group was fed with basal diet,and three treatment groups were added 0.1% tannin (tannin group),0.1% feeding cellulase (cellulase group) and 0.1% tannin +0.1% feeding cellulase (mixed group) in the basal diet respectively,3 replicates per group,each repeat 3 sheep.The test period was 70 days,in which the transition period was 7 days,the pre-test period was 7 days,and the positive feeding period was 56 days.At the end of the experiment,the rumen fluid of Hu sheep was collected,and the total DNA of the bacteria was extracted and PCR amplification was conducted.The amplified products were sequenced by Illumina Hi Seq 2500 for high-throughput sequencing.The results showed that:① A total of 957 440 pairs of microbial samples were obtained from 12 rumen samples,with an average of 55 997 clean tags,after filtering chimeras,a total of 502 965 effective tags were generated.AvgLen of rumen microbial samples from each group of sheep were between 419 and 420;②The Ace index and Chao1 index of rumen microorganisms in each group had no significant difference (P > 0.05);Compared with the control group,the Shannon index of rumen microorganisms in tannin,cellulose and mixed groups were significantly increased (P < 0.05).The Simpson index of rumen microorganisms in cellulase group was significantly lower than that in the control group (P < 0.05).③At the level of phylum:The abundance of Bacteroidetes of tannin group was significantly lower than that of the control group (P < 0.05);The abundance of the Firmicutes in tannin,cellulose and mixed group were lower than that of the control group,but the difference was not significant (P > 0.05).④On the genus level,the abundance of Prevotella_1 in tannin group was droped,and there was significant difference compared with the control and cellulase groups (P < 0.05),but there was no significant difference with mixed groups (P > 0.05).The abundance of Rumen-bacterium in tannin and cellulase groups were higher than that of the control and mixed group,and the difference with mixed group was significant (P < 0.05).In conclusion,adding tannin and cellulase at the same time could improve the diversity and abundance of rumen flora,affect the structure of rumen flora,and alleviate the inhibition of tannin on Bacteroidetes and Firmicutes.Mixed addition could alleviate the inhibition of cellulose decomposition by adding tannin alone.The dominant bacteria in the rumen of each group of Hu sheep were Bacteroidetes and Firmicutes.On the genus level,the dominant bacteria in the rumen of each group were studied Rikenellaceae_RC9_gut_group,Bacterium,Prevotella_1 and Rumen_bacterium.

Key words: tannin; feeding cellulase; Hu sheep; microbial flora; high-throughput sequencing

中图分类号:

S816.7

赵梦迪, 邸凌峰, 唐泽宇, 曹雪, 李成云. 单宁与饲用纤维素酶对湖羊瘤胃微生物菌群的影响[J]. 《中国畜牧兽医》, 2019, 46(1): 112-122.

ZHAO Mengdi, DI Lingfeng, TANG Zeyu, CAO Xue, LI Chengyun. Effects of Tannin and Feeding Cellulase on Rumen Microflora of Hu Sheep[J]. , 2019, 46(1): 112-122.

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