Untargeted Metabolomic Analysis of Gestationally Matched Human and Bovine Milk Samples at 2-Weeks Postnatal

Human breast milk is the ideal nutrition source for infant development during the first year of life. Epidemiological data demonstrates that bovine whole milk is often substituted for human milk during the first 12-months of life and may be associated with adverse infant outcomes. The goal of this project is to interrogate the human and bovine milk metabolome at 2-weeks postnatal to identify unique and overlapping metabolites that may impact infant health outcomes.Human milk ( = 10) was collected at 2-weeks postpartum from normal weight mothers (pre-pregnant BMI <25 kg/m2) that vaginally delivered term infants and planned to exclusively breastfeed for at least 2-months. Similarly, bovine milk ( = 10) was collected 2-weeks postpartum from normal weight primiparous Holstein dairy cows. Dairy cattle were housed in sand-bedded, shaded barns with access to fans and water soakers and fed a common transition cow total mixed ration. Untargeted metabolomics was completed on all milk samples using high-resolution mass spectrometry. Metabolomic analysis was implemented using an open-source containerized metabolomics pipeline. Data processing was completed using MZmine, mummichog and Python were used for statistical analysis.We detected 716 metabolomic features in human and bovine milk samples after quality control. Our analysis also revealed that 43% (312) of metabolomics features were present in both human and bovine milk, 23% (167) of metabolomics features were unique to human milk and 33% (237) of metabolomics features existed only in bovine milk samples. Pathway analysis revealed that sialic acid and glycosphingolipid metabolism ( < 0.0009) were common to human and bovine milk samples. We also found that amino acid (tryptophan, tyrosine, purine) metabolism ( < 0.005) was unique to bovine samples and vitamin B3 pathways ( = 0.03) was unique to human samples.Our analysis revealed a core milk metabolome shared between human and bovine samples. Collectively, these results highlight untargeted metabolomics as a potential strategy to identify unique and overlapping metabolites in bovine and human milk that may impact infant health outcomes.Research was supported by NIDDK/K01; SECIM P&F; CTSI Pilot Award; Robin Hood Foundation; NIH Loan Repayment Program.

人母乳是婴儿出生后第一年发育的理想营养来源。流行病学数据表明，在婴儿出生后的12个月内，牛乳常被用来替代母乳，并且可能与婴儿的不良后果有关。本项目的目标是在产后2周对人乳和牛乳的代谢组进行研究，以确定可能影响婴儿健康结果的独特和重叠的代谢物。从正常体重（孕前体重指数<25 kg/m²）、阴道分娩足月婴儿且计划至少纯母乳喂养2个月的母亲产后2周收集人乳（n = 10）。同样，从正常体重的初产荷斯坦奶牛产后2周收集牛乳（n = 10）。奶牛饲养在铺有沙子、有遮阳的牛舍中，可使用风扇和喷水器，并饲喂常见的过渡奶牛全混合日粮。使用高分辨率质谱法对所有乳样进行非靶向代谢组学研究。使用开源的容器化代谢组学流程进行代谢组学分析。使用MZmine进行数据处理，使用mummichog和Python进行统计分析。 经过质量控制后，我们在人乳和牛乳样本中检测到716种代谢组学特征。我们的分析还显示，43%（312种）的代谢组学特征在人乳和牛乳中均存在，23%（167种）的代谢组学特征为人乳所特有，33%（237种）的代谢组学特征仅存在于牛乳样本中。通路分析显示，唾液酸和鞘糖脂代谢（P < 0.0009）在人乳和牛乳样本中是共有的。我们还发现，氨基酸（色氨酸、酪氨酸、嘌呤）代谢（P < 0.005）为牛乳样本所特有，维生素B3通路（P = 0.03）为人乳样本所特有。 我们的分析揭示了人乳和牛乳样本之间共享的核心乳代谢组。总体而言，这些结果强调了非靶向代谢组学作为一种潜在策略，可用于确定牛乳和人乳中可能影响婴儿健康结果的独特和重叠的代谢物。 本研究由美国国家糖尿病、消化和肾脏疾病研究所/ K01项目；东南临床与转化医学研究所（SECIM）规划与设施项目；临床与转化科学研究所（CTSI）试点奖；罗宾汉基金会；美国国立卫生研究院贷款偿还计划资助。