研究论文 RESEARCH PAPER

不同乳脂率奶牛瘤胃微生物及其代谢物差异分析

  • 沈子亮 ,
  • 王全 ,
  • 曹辉 ,
  • 胡立萍 ,
  • 田雨 ,
  • 姜惠 ,
  • 毛永江 ,
  • 杨章平 ,
  • 张慧敏
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  • 1. 扬州大学动物科学与技术学院, 扬州 225009;
    2. 姜堰区白米镇畜牧兽医站, 泰州 225505;
    3. 南京卫岗乳业有限公司, 南京 210008
沈子亮(1997—),男,江苏泰州人,硕士研究生,畜牧专业。E-mail:3143197661@qq.com

收稿日期: 2021-10-04

  网络出版日期: 2022-05-14

基金资助

江苏省研究生科研与实践创新计划项目(SJCX21_1627);国家自然科学基金面上项目(31972555);江苏省现代农业(奶牛)产业技术体系(JATS[2021]486);扬州大学大学生科技创新基金(X20210671)

Differential Analysis of Rumen Microbes and Their Metabolites for Dairy Cows with Different Milk Fat Percentages

  • SHEN Ziliang ,
  • WANG Quan ,
  • CAO Hui ,
  • HU Liping ,
  • TIAN Yu ,
  • JIANG Hui ,
  • MAO Yongjiang ,
  • YANG Zhangping ,
  • ZHANG Huimin
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  • 1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
    2. Baimi Animal Husbandry and Veterinary Station in Jiangyan District, Taizhou 225505, China;
    3. Nanjing Weigang Dairy Co., Ltd., Nanjing 210008, China

Received date: 2021-10-04

  Online published: 2022-05-14

摘要

本研究旨在分析不同乳脂率奶牛瘤胃微生物及其代谢物的差异。本试验基于江苏省某奶牛场2020年的中国荷斯坦奶牛生产性能测定(DHI)数据,选取胎次、泌乳天数及产奶量相近的高乳脂率奶牛(H组)及低乳脂率奶牛(L组)各5头,收集瘤胃液及奶样,测定常规乳成分指标、瘤胃液挥发性脂肪酸浓度与消化酶活性;通过16S rDNA测序和非靶向液相色谱-质谱(LC-MS)代谢组学技术分析瘤胃微生物组成及其代谢物。结果显示:相同饲粮条件下,H组和L组的乳脂率分别为5.82%和3.60%,二者差异极显著(P<0.01)。H组瘤胃液乙酸/丙酸比值显著高于L组(P<0.05)。16S rDNA测序结果表明瘤胃内厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和变形菌门(Proteobacteria)为优势菌门,且H组Kiritimatiellaeota的相对丰度显著低于L组(P<0.05)。在属水平上,普雷沃氏菌属1(Prevotella_1)、解琥珀酸弧菌属(Succiniclasticum)和琥珀酸弧菌科UCG-001(Succinivibrionaceae_UCG-001)为优势菌属。2组间奶牛瘤胃菌群的丰富度与多样性无显著差异(P>0.05),但菌群组成结构存在较大差异。H组奶牛瘤胃内嗜木聚糖真杆菌群(Eubacterium_xylanophilum_group)的相对丰度显著高于L组(P<0.05),而克里斯滕森菌科R-7群(Christensenellaceae_R-7_group)、产粪甾醇真杆菌群(Eubacterium_coprostanoligenes_group)、瘤胃球菌科UCG-010(Ruminococcaceae_UCG-010)、丁酸弧菌属2(Butyrivibrio_2)、产甲烷短杆菌属(Methanobrevibacter)、p-1088-a5_gut_group、聚乙酸菌属(Acetitomaculum)、毛螺梭菌属10(Lachnoclostridium_10)、毛螺旋菌科ND3007群(Lachnospiraceae_ND3007_group)的相对丰度显著低于L组(P<0.05)。H组瘤胃内木聚糖酶活性显著高于L组(P<0.05),但脂肪酶活性极显著低于L组(P<0.01)。代谢组学分析共发现了4种差异代谢物[差异倍数(FC)≥1.5且P<0.05],能够有效区分L组和H组。与L组相比,H组内显著上调的差异代谢物有月桂酸、琥珀酸、月桂酸乙酯及12,13-二羟基十八碳烯酸(P<0.05),这些代谢物富集在脂肪酸生物合成及氨基酸代谢等通路。此外,瘤胃微生物与代谢组数据关联分析显示,月桂酸、月桂酸乙酯、二羟基十八烯酸均与粪球菌属2(Coprococcus_2)呈显著正相关(P<0.05),琥珀酸与琥珀酸弧菌科UCG-002(Succinivibrionaceae_UCG-002)和普雷沃氏菌科Ga6A1群(Prevotellaceae_Ga6A1_group)呈显著正相关(P<0.05)。综上所述,相同饲粮条件下,乳脂率不同的奶牛,其瘤胃发酵模式、微生物组成及其代谢物均有不同,这些微生物的变化可引起瘤胃内脂肪酸生物合成、氨基酸代谢等通路的改变,最终影响乳脂率。

本文引用格式

沈子亮 , 王全 , 曹辉 , 胡立萍 , 田雨 , 姜惠 , 毛永江 , 杨章平 , 张慧敏 . 不同乳脂率奶牛瘤胃微生物及其代谢物差异分析[J]. 动物营养学报, 2022 , 34(5) : 3000 -3011 . DOI: 10.3969/j.issn.1006-267x.2022.05.029

Abstract

The objective of this study was to analyze the difference of rumen microbes and their metabolites for dairy cows with different milk fat percentages. Five high milk fat percentage dairy cows (H group) and five low milk fat percentage dairy cows (L group) with similar parity, lactation days and milk yield were selected from a dairy farm in Jiangsu province based on 2020 dairy herd improvement (DHI) data of Chinese Holstein dairy cows. Ruminal fluid and milk samples were collected and used to determine ruminal fluid volatile fatty acid (VFA) concentrations and digestive enzyme activities, conventional milk composition indexes, respectively. Rumen microbial composition and rumen metabolites were analyzed by 16S rDNA sequencing and untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics techniques. The results showed as follows:under the same diet condition, the milk fat percentage in H group and L group were 5.82% and 3.60%, respectively, and the difference between them was extremely significant (P<0.01). And ruminal fluid acetate/propionate ratio in H group was significantly higher than that in L group (P<0.05). The results of 16S sequencing indicated that Firmicutes, Bacteroidetes and Proteobacteria were the dominant phylum in the rumen of dairy cows, the relative abundance of Kiritimatiellaeota in H group was significantly decreased compared with the L group (P<0.05). At genus level, Prevotella_1, Succiniclasticum and Succinivibrionaceae_UCG-001 were the dominant genera. There was no significant difference in the richness and diversity of rumen bacteria between the two groups (P>0.05), but the rumen bacterial composition structure between the two groups were different. The relative abundance of Eubacterium_xylanophilum_group in H group was significantly higher than that in L group (P<0.05), while the relative abundances of Christensenellaceae_R-7_group, Eubacterium_coprostanoligenes_group, Ruminococcaceae_UCG-010, Butyrivibrio_2, Methanobrevibacter, p-1088-a5_gut_group, Acetitomaculum, Lachnoclostridium_10, Lachnospiraceae_ND3007_group were significantly lower than those in L group (P<0.05). Compared with the L group, ruminal fluid xylanase activity in H group was significantly increased (P<0.05), but the lipase activity was extremely significantly decreased (P<0.01). Metabolome analysis found a total of 4 different metabolites (fold change ≥ 1.5 and P<0.05), which could effectively distinguish L group from H group. Compared with the L group, lauric acid, succinic acid, lauric acid ethyl ester and 12,13-dihydroxy-9Z-octadecenoic acid (12,13-DiHOME) were significantly up-regulated in H group (P<0.05). These metabolites were enriched in fatty acid biosynthesis and amino acid metabolism pathways. In addition, the correlation analysis of rumen microbes and metabolites showed that lauric acid, lauric acid ethyl ester and 12,13-DiHOME were significantly positively correlated with Coprococcus_2 (P<0.05). Succinic acid was significantly positively correlated with Succinivibrionaceae_UCG-002 and Prevotellaceae_Ga6A1_group (P<0.05). In conclusion, under the same diet condition, the rumen fermentation pattern, microbial composition and their metabolites are different of dairy cows with different milk fat percentages. These differential microbes can cause the changes of fatty acid biosynthesis and amino acid metabolism pathways in rumen, then affect milk fat percentage.

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