不同年龄奶水牛瘤胃菌群多样性研究

彭科兰; 任大喜; 刘建新

doi:10.3969/j.issn.1006-267x.2019.11.020

动物营养学报 >

2019 , Vol. 31 >Issue 11: 5053 - 5064

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.11.020

反刍与草食动物营养 Ruminant and herbivore nutrition

不同年龄奶水牛瘤胃菌群多样性研究

彭科兰 ,
任大喜 ,
刘建新

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浙江大学动物科学学院奶业科学研究所, 杭州 310058

彭科兰(1990-),女,湖南邵阳人,硕士研究生,从事动物消化道微生物研究。E-mail:pengkelan@live.cn

收稿日期: 2019-04-16

网络出版日期: 2019-11-19

基金资助

国家自然科学基金项目（31328022）

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Diversity of Rumen Microflora in Dairy Buffaloes at Different Ages

PENG Kelan ,
REN Daxi ,
LIU Jianxin

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Institute of Dairy Science, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Received date: 2019-04-16

Online published: 2019-11-19

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摘要

本研究旨在利用16S rRNA高通量测序技术分析不同年龄奶水牛瘤胃菌群多样性，探究其随年龄增长变化的规律。选择4个年龄组，即Y组（12月龄，胎次=0）、M组（3~5岁，胎次=1）、E组（6~8岁，胎次=2）和O组（9岁及以上，胎次≥3），各组奶水牛分别为9、7、4和5头，采集瘤胃液并提取总DNA，经PCR扩增后对16S rDNA的V3~V4区进行高通量测序。结果表明：alpha多样性分析结果发现，随着年龄增长，奶水牛瘤胃菌群数量呈现显著下降的趋势（P=0.07），菌属相对丰度显著减少（P<0.05）；beta多样性分析发现，不同年龄奶水牛的瘤胃菌群结构存在明显的不同，并随年龄增加个体间的瘤胃菌群结构差异逐渐增大。门水平下，本试验所有年龄奶水牛瘤胃的优势菌群为拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）和变形菌门（Proteobacteria）；属水平下，普雷沃氏菌属_1（Prevotella_1）相对丰度在4个年龄组均最高，优势菌属和次级优势菌属的相对丰度随奶水牛年龄的增长呈现相互聚拢的趋势。不同年龄奶水牛瘤胃中存在着差异显著的特征性菌属，12月龄奶水牛瘤胃中以乳酸菌属（Lactobacillus）和糖酵菌属（Saccharofermentans）为主，其相对丰度随着年龄增长而显著减少（P<0.05）；3~5岁奶水牛瘤胃中则以未分类梭菌目菌属（unclassified_o_Clostridiales）和未排位双歧杆菌科菌属（norank_f_Bifidobacteriaceae）为主，相对丰度显著高于12月龄和9岁及以上年龄（P<0.05）；9岁及以上年龄奶水牛瘤胃中的未分类普雷沃氏菌科菌属（unclassified_f_Prevotellaceae）和候选单胞生糖菌属（Candidatus_Saccharimonas）相对丰度高于12月龄和3~5岁。共性网络物种关联分析结果显示，奶水牛瘤胃菌群间的相关性和紧密性随着年龄增加而逐渐下降。综上所述，随着奶水牛年龄增长，其瘤胃菌群数量和相对丰度显著减少，优势菌群和亚优势菌群呈现一定的年龄特征变化；奶水牛瘤胃存在着一些年龄特征菌属，其菌群结构紧密性随年龄增加而减弱，表明宿主年龄增长可影响瘤胃菌群结构和功能的稳定。

关键词： 奶水牛; 瘤胃; 微生物多样性; 菌群; 年龄

本文引用格式

彭科兰 , 任大喜 , 刘建新 . 不同年龄奶水牛瘤胃菌群多样性研究[J]. 动物营养学报, 2019 , 31(11) : 5053 -5064 . DOI: 10.3969/j.issn.1006-267x.2019.11.020

Abstract

This experiment was conducted to study the diversity of rumen microflora in dairy buffaloes at different ages using 16S rRNA high throughput sequencing, and to explore the charge with age increasing. Four age groups were group Y (12 months of age, parity=0), group M (3 to 5 years of age, parity=1), group E (6 to 8 years of age, parity=2), and group O (more than 9 years of age, parity ≥ 3), and had 9, 7, 4 and 5 dairy buffaloes in groups, respectively. Total bacterial DNA was extracted from the rumen fluids. After PCR amplication, V3 to V4 regions of 16S rDNA were sequenced by high-throughput sequencing technology. The results showed as follows:alpha diversity analysis indicated that the number of rumen bacteria tended to decrease with age increasing (P=0.07), and the relative abundance decreased significantly (P<0.05). Beta diversity analysis indicated that significant differences were found in the rumen bacterial structure among dairy buffalo at different ages. At the phylum level, the dominant bacteria for all ages were Bacteroidetes, Firmicutes and Proteobacteria. At the phylum level, the relative abundance of Prevotella_1 was the highest among the four age groups. The relative abundance of dominant and sub-dominant genus tended to close with each other with age increasing. There existed some significant characteristic rumen bacteria at species level among different ages. The Lactobacillus and Saccharofermentans were the main bacterial genus in rumen of dairy buffalo at 12 months of age, and the relative abundance decreased significantly with age increasing (P<0.05); the unclassified_o_Clostridiales and g_norank_f_Bifidobacteriaceae were the main bacterial genus in rumen of dairy buffalo at 3 to 5 years of age, and the relative abundance were significantly higher than those of dairy buffalo at 12 months of age and 9 years of age and above (P<0.05); the relative abundance of g_unclassified_f_Prevotellaceae and g_Candidatus_Saccharimonas were higher than those at 12 months of age and 3 to 5 days of age. Co-occurrence analysis indicated that the correlation and compactness of the rumen bacteria in dairy buffalo were gradually decreased with age increasing. From the results obtained in this work, it is indicated that the rumen bacteria number and relative abundance in dairy buffalo decrease with age increasing. There exist some characteristic rumen bacteria at different ages. The correlation among the rumen bacteria decreases gradually with age increasing, suggesting that the stability of rumen microflora may reduce with the increasing age of dairy buffalo.

Key words： dairy buffalo; rumen; bacterial diversity; bacteria; age

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