Biomarker and pathway analyses of urine metabolomics dairy cows when corn stover replaces alfalfa hay

Huizeng Sun; Bing Wang; Jiakun Wang; Hongyun Liu; Jianxin Liu

摘要

Background:Alfalfa hay and corn stover are different type of forages which can significantly impact a cow's lactation performance,but the underlying metabolic mechanism has been poorly studied.We used biomarker and pathway analyses to characterize related biomarkers and pathways based on urine metabolomics data from different forage treatments.Urine was collected from 16 multiparous Holstein dairy cows fed alfalfa hay (AH,high-quality forage,n =8) and corn stover (CS,low-quality forage,n =8) respectively.Gas chromatography-time of flight/mass spectrometry (GC-TOF/MS) was performed to identify metabolites in urine and the metaboanalyst online platform was used to do biomarker and pathway analysis.Results:Hippuric acid (HUA) and N-methyl-glutamic (NML-Glu) indicated the most significant difference between the two diets,when statistically validated by biomarker analysis.HUA was also validated by standard compound quantitative method and showed significant higher concentration in CS group than AH group (2.8282 vs.0.0005 mg/mL;P ＜ 0.01).The significant negative correlation between milk yield and HJA (R2=0.459;P ＜ 0.01) and significant positive correlation between milk yield and NML-Glu (R2 =0.652;P ＜ 0.01) were characterized.The pathway analysis revealed that these different metabolites were involved in 17 pathways including 7 influential pathways (pathway impact value ＞ 0):Tyr metabolism,starch and sucrose metabolism,amino sugar and nucleotide sugar metabolism,galactose metabolism,Phe,Tyr and Try biosynthesis,purine metabolism,and glycerolipid metabolism.Based on the metabolome view map,the Phe,Tyr and Try biosynthesis pathway exhibited the highest impact value (0.50),and the Holm-Bonferroni multiple testing-based analysis revealed the most significant difference in the Tyr metabolism pathway (Holm P =0.048).Conclusions:The identified HUA and NML-Glu may serve as potential biomarkers for discriminating CS and AH diets and could be used as candidates for milk yield related mechanistic investigations.Integrated network pathways associated with related metabolites provide a helpful perspective for discovering the effectiveness of forage quality in lactation performance and provides novel insights into developing strategies for better utilization of CS and other low-quality forage in China.

机译：背景：苜蓿干草和玉米秸秆是不同类型的草料，可以显着影响奶牛的泌乳性能，但其潜在的代谢机制尚未得到充分研究。分别从16只饲喂苜蓿干草（AH，优质牧草，n = 8）和玉米秸秆（CS，低品质牧草，n = 8）的荷斯坦奶牛中收集尿液。质谱法（GC-TOF / MS）鉴定尿液中的代谢物，并使用代谢物分析仪在线平台进行生物标志物和路径分析。结果：表明尿酸（HUA）和N-甲基谷氨酸（NML-Glu）当通过生物标志物分析进行统计学验证时，两种饮食之间的差异最为显着。HUA还通过标准化合物定量方法进行了验证，并且在CS组中显示出明显较高的浓度比AH组（2.8282 vs.0.0005 mg / mL; P ＜0.01）。牛奶产量与HJA呈显着负相关（R2 = 0.459; P ＜0.01），牛奶产量与NML-Glu呈显着正相关（R2 = 0.652）。 ; P ＜0.01）。途径分析表明，这些不同的代谢物涉及17种途径，包括7种影响途径（途径影响值＞ 0）：酪氨酸代谢，淀粉和蔗糖代谢，氨基糖和核苷酸糖代谢，半乳糖代谢，Phe，Tyr和Try生物合成，嘌呤代谢和甘油脂代谢。基于代谢组图，Phe，Tyr和Try生物合成途径表现出最高的影响值（0.50），并且基于Holm-Bonferroni多重测试的分析显示结论：经鉴定的HUA和NML-Glu可能成为区分CS和AH日粮的潜在生物标志物，并且可以用作牛奶产量相关的候选物。与相关代谢产物相关的综合网络途径为发现饲草质量对泌乳性能的有效性提供了有益的见解，并为中国更好地利用CS和其他劣质饲草的发展战略提供了新的见解。

Biomarker and pathway analyses of urine metabolomics dairy cows when corn stover replaces alfalfa hay

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