Untargeted metabolomic investigate milk and ruminal fluid of Holstein cows supplemented with Perilla frutescens leaf

Milk compounds are important for human nutrient requirements and health. The ruminal metabolic profile is responsible for dietary nutrition and determines milk production. Perilla frutescens leaf (PFL) is a commonly used medicinal herb due to its bioactive metabolites. This study elucidated the effects of PFL on the metabolome of two biofluids (rumen fluid and milk) of 14 cows fed a basic total mixed ration diet (CON, n = 7) and supplemented with 300 g/d PFL per cow (PFL, n = 7) by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. Milk PE-NMe (18:1(9Z)/18:1(9Z)) and DG (18:0/20:4(5Z,8Z,11Z,14Z)/0:0), oleanolic acid, and nucleotides were upregulated, and milk medium-chain fatty acids (2-hydroxycaprylic acid) were down-regulated in response to PFL. The supplementation of PFL increased the abundance of pyrimidine nucleotides both in rumen fluid and milk. The pathways of pyrimidine metabolism and biosynthesis of unsaturated fatty acids were enriched both in the rumen fluid and milk. We also found the milk 2-hydroxycaprylic acid was positively correlated with ruminal uridine 5-monophosphate, and was negatively correlated with ruminal deoxycytidine, and the milk thymidine was positively correlated with ruminal icosenoic acid. This study found that the supplementation of PFL could alter the ruminal metabolic profiles and milk synthesis through regulation of the pathways of pyrimidine metabolism and biosynthesis of unsaturated fatty acids. Our new findings provide comprehensive insights into the metabolomics profile of rumen fluid and milk, supporting the potential production of Perilla frutescens milk in dairy cows.