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荷斯坦牛高、低乳脂率牛乳代谢组分析
引用本文:王影,文亮,母童,冯小芳,刘佳敏,张娟,温万,顾亚玲.荷斯坦牛高、低乳脂率牛乳代谢组分析[J].畜牧兽医学报,2022,53(5):1396-1408.
作者姓名:王影  文亮  母童  冯小芳  刘佳敏  张娟  温万  顾亚玲
作者单位:1. 宁夏大学农学院, 银川 750021;2. 宁夏农垦贺兰山奶业有限公司, 银川 750000;3. 宁夏畜牧工作站, 银川 750001
基金项目:宁夏自治区育种专项“优质高产奶牛选育资助项目”(2019NYYZ05)
摘    要:旨在筛选牛乳不同乳脂水平潜在标志性差异代谢物。本研究中乳样采自宁夏农垦贺兰山奶业茂盛牧场,采集体重与月龄相近且健康的头胎次荷斯坦牛泌乳中后期(180~210 d)的早、中、晚班次新鲜乳样(4∶3∶3混合)245份。DHI测定后筛选体细胞数低于20万个·mL-1,乳脂率具有极端差异的样品26份,其中乳脂率大于4.4%为高乳脂率组(HF组,13份),乳脂率小于3.0%为低乳脂率组(LF组,13份)。之后采用超高效液相色谱-质谱法(UHPLC-MS)对样品进行代谢组学分析。偏最小二乘法-判别分析(PLS-DA)结果显示,相对于LF组,HF组代谢轮廓发生了变化,在HF与LF两组中共鉴定出343种代谢产物,根据VIP值(VIP>1)和单变量统计分析(P<0.05),正、负离子模式下共筛选出60种显著差异代谢物(P<0.05);其中依据P<0.5且FC>2.0及P<0.5且FC<0.5,HF组奶样中15种代谢物含量高于LF组,6种代谢物含量低于LF组;进一步通过ROC曲线下面积AUC>0.85,发现有18种差异代谢物对两组牛乳具有显著区别能力,即:N-油酰甘氨酸、皮质醇、δ-生育酚、PC(14∶0e/9∶0)。结果发现,不同乳脂水平显著差异代谢物有60种,且代谢物N-油酰甘氨酸、皮质醇、δ-生育酚、PC(14∶0e/9∶0)可作为高乳脂率牛乳潜在标志性差异代谢物。本研究结果为奶牛乳脂合成提供了有价值的理论依据,同时也为研究奶牛的泌乳过程提供了资料。

关 键 词:荷斯坦奶牛  乳脂率  代谢组学  标志代谢物  
收稿时间:2021-10-12

Metabolomic Analysis of Milk from Holstein Cows with High and Low Milk Fat Percentage
WANG Ying,WEN Liang,MU Tong,FENG Xiaofang,LIU Jiamin,ZHANG Juan,WEN Wan,GU Yaling.Metabolomic Analysis of Milk from Holstein Cows with High and Low Milk Fat Percentage[J].Acta Veterinaria et Zootechnica Sinica,2022,53(5):1396-1408.
Authors:WANG Ying  WEN Liang  MU Tong  FENG Xiaofang  LIU Jiamin  ZHANG Juan  WEN Wan  GU Yaling
Institution:1. School of Agriculture, Ningxia University, Yinchuan 750021, China;2. Ningxia Agriculture Helanshan Dairy Corporation Ltd., Yinchuan 750000, China;3. Animal Husbandry Extension Station, Yinchuan 750001, China
Abstract:The study aimed to screen the potential landmark differential metabolites in milk with different milk fat levels. In this study, 245 fresh milk samples (4:3:3 mixture) were collected from the early, middle and late shifts of healthy first-calf Holstein cows with similar weight and age in the middle and late stages of lactation (180-210 d) from the Maosheng pasture of Helanshan in Ningxia state farm. After DHI determination, 26 samples with the number of somatic cells less than 200 000 pcs·mL-1 and extreme differences in milk fat percentage were screened. Among them, the milk with higher milk fat percentage than 4.4% was the high milk fat percentage group (HF group, 13 samples), and the milk with less milk fat percentage than 3.0% was the low milk fat percentage group (LF group, 13 samples). The samples were then subjected to metabolomic analysis using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS). The results of PLS-DA showed that, compared with LF group, the metabolic profile of HF group changed, and 343 metabolites were identified in HF and LF groups. According to VIP value (VIP>1) and univariate statistical analysis (P<0.05), 60 significantly differential metabolites were screened out in positive and negative ion modes (P<0.05). Among them, according to P<0.5 and FC>2.0 and P<0.5 and FC<0.5, the content of 15 metabolites in milk samples of HF group was higher than that of LF group, and the content of 6 metabolites was lower than that of LF group.Further, through ROC curve, it was found that there were 18 kinds of differential metabolites which had remarkable distinguishing ability to the two groups of milk, among which the differential metabolites with AUC>0.85 were N-oleoylglycine, cortisol, δ-tocopherol and PC (14:0e/9:0). The results showed that there were 60 metabolites with significant differences in different milk fat levels, and the metabolites N-oleoylglycine, cortisol, δ-tocopherol and PC (14:0e/9:0) could be used as potential landmark metabolites of high milk fat percentage. The results of this study provide valuable theoretical basis for milk fat synthesis of dairy cows, and also provide data for studying the lactation process of dairy cows.
Keywords:Holstein cow  milk fat percentage  metabolomics  marker metabolite  
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