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浙江大学学报(农业与生命科学版)  2023, Vol. 49 Issue (4): 578-590    DOI: 10.3785/j.issn.1008-9209.2022.06.291
动物科学与动物医学     
泌乳中期荷斯坦奶牛瘤胃细菌群落组成与多样性变化
马晓娇(),薛茗元,孙会增,刘建新()
浙江大学动物科学学院奶业科学研究所,浙江 杭州 310058
Changes in the composition and diversity of the rumen bacterial community in mid-lactation Holstein cows
Xiaojiao MA(),Mingyuan XUE,Huizeng SUN,Jianxin LIU()
Institute of Dairy Science, College of Animal Sciences, Zhejiang University, Hangzhou 310058, Zhejiang, China
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摘要:

本研究旨在探究泌乳中期荷斯坦奶牛瘤胃细菌群落组成及其多样性的变化。共设置2个试验:试验1,选取20头泌乳中期健康高产荷斯坦奶牛,饲喂相同饲粮,持续8周,分别在第0、4、7周的第7天采集瘤胃内容物;试验2,选取30头泌乳中期健康高产荷斯坦奶牛,在基础饲粮的基础上补充20 g/d瘤胃保护蛋氨酸,持续8周,分别在第0、8周的第7天采集瘤胃内容物。分析2个试验在不同时间点采集到的瘤胃内容物的细菌群落组成与多样性变化,以及功能稳定性差异。结果发现:2个试验中,瘤胃细菌的α多样性、β多样性、功能稳定性在不同时间点之间都不存在显著差异(P>0.05)。试验1在高丰度细菌的门和属水平上分别发现6个与1个差异细菌,分别是放线菌门、变形菌门、蓝菌门、绿弯菌门、互养菌门、纤维杆菌门与Lachnospiraceae_NK3A20_group;试验2在高丰度细菌的门和属水平上分别发现3个与1个差异细菌,分别为放线菌门、螺旋体门、迷踪菌门与unclassified_f_Lachnospiraceae。由此可见,在相同的饲粮条件下,泌乳中期荷斯坦奶牛瘤胃细菌组成随时间变化的程度有限,其多样性与功能稳定性均未发生显著改变;在基础饲粮基础上补充瘤胃保护蛋氨酸时,得到的结果相似。综上所述,本研究表明,处于泌乳中期荷斯坦奶牛的瘤胃微生物群落组成和功能相对稳定,在奶牛泌乳中期开展相关试验时,无需特别考虑瘤胃细菌会随泌乳阶段进程出现群落结构和功能的改变。

关键词: 荷斯坦奶牛泌乳中期瘤胃细菌微生物多样性功能稳定性    
Abstract:

This study aimed to investigate the changes in the composition and diversity of the rumen bacterial community in mid-lactation Holstein cows. In trial 1, a total of 20 healthy high-yielding Holstein cows at mid-lactation were continuously fed with the same basal diet for eight weeks and the rumen contents were collected on the 7th day at weeks zero, four, and seven. In trial 2, a total of 30 healthy high-yielding Holstein cows at mid-lactation were supplemented with 20 g/d rumen-protected methionine to the basal diet, and the rumen contents were collected on the 7th day at weeks zero and eight. The rumen contents collected at different time points were analyzed for changes in composition and diversity of the bacterial community, as well as differences in functional stability in both trials. The results showed that there were no significant differences in the alpha diversity, beta diversity, and functional stability of rumen bacteria at different time points (P>0.05) in both trials. In trial 1, six and one differential bacteria, such as Actinobacteriota, Proteobacteria, Cyanobacteria, Chloroflexi, Synergistota, and Fibrobacterota, as well as Lachnospiraceae_NK3A20_group, were found in highly abundant bacteria at phylum and genus levels, respectively. In trial 2, three and one differential bacteria, such as Actinobacteriota, Spirochaetota, and Elusimicrobiota, as well as unclassified_f_Lachnospiraceae, were found in highly abundant bacteria at phylum and genus levels, respectively. It is indicated that the rumen bacterial community composition of Holstein cows at mid-lactation changed to a limited extent over time under the same feeding conditions, with no significant changes in their diversity and functional stability. Similar results were obtained when supplemented with rumen-protected methionine to the basal diet. In conclusion, the results suggested that the rumen microbial community composition and function of Holstein cows at mid-lactation were relatively stable, and there is no need to specifically consider the changes in rumen bacterial community structure and function along with lactation progress in the related experiments developed during the mid-lactation.

Key words: Holstein dairy cows    mid-lactation    rumen bacteria    microbial diversity    functional stability
收稿日期: 2022-06-29 出版日期: 2023-08-29
CLC:  S823  
基金资助: 国家自然科学基金项目(31872380)
通讯作者: 刘建新     E-mail: 22017022@zju.edu.cn;liujx@zju.edu.cn
作者简介: 马晓娇(https://orcid.org/0000-0002-5352-8939),E-mail:22017022@zju.edu.cn
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引用本文:

马晓娇,薛茗元,孙会增,刘建新. 泌乳中期荷斯坦奶牛瘤胃细菌群落组成与多样性变化[J]. 浙江大学学报(农业与生命科学版), 2023, 49(4): 578-590.

Xiaojiao MA,Mingyuan XUE,Huizeng SUN,Jianxin LIU. Changes in the composition and diversity of the rumen bacterial community in mid-lactation Holstein cows. Journal of Zhejiang University (Agriculture and Life Sciences), 2023, 49(4): 578-590.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2022.06.291        https://www.zjujournals.com/agr/CN/Y2023/V49/I4/578

参数

Parameter

试验1 Trial 1试验2 Trial 2

第0周

Week

zero

第4周

Week

four

第7周

Week

seven

均值

标准误

SEM

p

p-value

第0周

Week

zero

第8周

Week

eight

均值

标准误

SEM

p

p-value

泌乳天数 Lactation day/d190218239141197
干物质采食量 DMI/(kg/d)25.626.024.30.370.1524.9a25.7b0.420.05
奶产量 Milk yield/(kg/d)
牛奶 Milk (M)36.4a34.6ab32.7b0.600.0435.435.60.430.61
乳脂矫正乳 FCM38.936.434.80.770.0738.4b40.6a0.810.01
乳成分 Milk composition
乳蛋白 Milk protein/%3.323.363.340.0260.763.52a3.46b0.0240.03
乳脂 Milk fat/%3.933.833.890.0800.904.03b4.37a0.110<0.01
乳糖 Milk lactose/%4.964.935.010.0200.305.115.130.0210.45
总固形物 Total solids/%12.9012.6712.790.0820.5512.90b13.22a0.1180.01
乳中尿素氮 MUN/(mg/dL)13.5ab14.1a12.3b0.300.0416.916.60.600.62
体细胞数 SCC/(103 mL-1)84.343.883.922.530.7168.057.714.020.47
饲料效率 Feed efficiency
M/DMI1.421.331.350.0220.211.421.390.0450.08
FCM/DMI1.521.401.420.0260.151.531.570.0630.90
表1  试验奶牛生产性能

参数

Parameter

第0周

Week zero

第8周

Week eight

均值标准误

SEM

p

p-value

pH6.49a6.40b0.0350.02
NH3-N/(mg/dL)16.314.40.980.06
总挥发性脂肪酸 Total VFAs/(mmol/L)117.3120.04.540.56
各脂肪酸摩尔分数 Mole fraction of each fatty acid/%
乙酸 Acetate61.862.50.7840.38
丙酸 Propionate22.823.70.600.15
丁酸 Butyrate11.4a10.8b0.21<0.01
异丁酸 Isobutyrate1.20.80.330.24
戊酸 Valerate1.4a1.1b0.05<0.01
异戊酸 Isovalerate1.3a1.2b0.05<0.01
乙酸/丙酸 Acetate/propionate2.732.720.1300.95
表2  奶牛瘤胃发酵指标(试验2)

参数

Parameter

试验1 Trial 1试验2 Trial 2

第0周

Week zero

第4周

Week four

第7周

Week seven

均值标准误

SEM

p

p-value

第0周

Week zero

第8周

Week eight

均值标准误

SEM

p

p-value

Sobs指数

Sobs index

164.00171.90168.602.6920.58176.77169.606.0060.94

Shannon指数

Shannon index

2.843.003.140.0650.213.063.000.0530.83

Simpson指数

Simpson index

0.190.160.130.0120.150.140.160.0110.98

ACE指数

ACE index

166.90175.20172.902.7160.58181.67173.236.5720.91

Chao 1指数

Chao 1 index

166.40174.50172.002.6690.63181.03172.866.5910.81
表3  瘤胃细菌 α 多样性指数
图1  瘤胃细菌属水平的 β 多样性(试验1)
图2  瘤胃细菌在不同时间点之间的差异(试验1)门(A)和属(B)水平前15名高丰度微生物在不同时间点之间的差异分析;C. LEfSe多级物种差异判别分析;D.差异微生物的LDA得分分布柱状图(LDA得分>2)。*、**、***分别表示在P<0.05、P<0.01、P<0.001水平差异有统计学意义。
图3  瘤胃细菌区系中衰减值和缓冲值的时间变化(试验1)
图4  瘤胃细菌属水平的 β 多样性(试验2)
图5  瘤胃细菌在不同时间点之间的差异(试验2)门(A)和属(B)水平前15名高丰度微生物在不同时间点之间的差异分析;C. LEfSe多级物种差异判别分析;D. 差异微生物的LDA得分分布柱状图(LDA得分>3)。*、**分别表示在P<0.05、P<0.01水平差异有统计学意义。
图6  瘤胃细菌区系中衰减值和缓冲值的时间变化(试验2)
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