Metabolic Profiling of Rumen Fluid and Milk in Lactating Dairy Cattle Influenced by Subclinical Ketosis Using Proton Nuclear Magnetic Resonance Spectroscopy

SIMPLE SUMMARY: Ketosis metabolic research is extremely rare in Korea. This study aimed to compare the rumen fluid and milk metabolites between healthy and subclinical ketosis-diagnosed lactating dairy cattle. Six Holstein cows were allocated into two groups based on whether they fit the criteria for subclinical ketosis, and their rumen fluid and milk samples were collected from the stomach tube and pipeline milking system. Rumen fluid and milk samples metabolites were analyzed using proton nuclear magnetic resonance spectroscopy. They were identified and quantified using the Chenomx NMR Suite 8.4 software and statistical analysis was performed using Metaboanalyst 5.0. In rumen fluid, ruminant energy source metabolites (acetate, glucose, and propionate) were significantly higher in the healthy group, whereas in milk, ketone body metabolites (3-hydroxybutyrate and acetoacetate) were significantly higher in the subclinical ketosis-diagnosed group. This report will serve as a reference guide for future studies on ketosis metabolomics in Korea. ABSTRACT: Ketosis metabolic research on lactating dairy cattle has been conducted worldwide; however, there have been very few Korean studies. Biofluids from lactating dairy cattle are necessary to study ketosis metabolic diseases. Six Holstein cows were divided into two groups (healthy (CON) and subclinical ketosis diagnosed (SCK)). Rumen fluid and milk samples were collected using a stomach tube and a pipeline milking system, respectively. Metabolites were determined using proton nuclear magnetic resonance (NMR) spectroscopy and they were identified and quantified using the Chenomx NMR Suite 8.4 software and Metaboanalyst 5.0. In the rumen fluid of the SCK group, butyrate, sucrose, 3-hydroxybutyrate, maltose, and valerate levels were significantly higher than in the CON group, which showed higher levels of N,N-dimethylformamide, acetate, glucose, and propionate were significantly higher. Milk from the SCK group showed higher levels of maleate, 3-hydroxybutyrate, acetoacetate, galactonate, and 3-hydroxykynurenine than that from the CON group, which showed higher levels of galactitol, 1,3-dihydroxyacetone, γ-glutamylphenylalanine, 5-aminolevulinate, acetate, and methylamine. Some metabolites are associated with ketosis diseases and the quality of rumen fluid and milk. This report will serve as a future reference guide for ketosis metabolomics studies in Korea.