BackgroundThe metabolic processes of cows undergo significant changes during subclinical mastitis, but their molecular mechanisms have not been clearly elucidated. This study investigated the changes in milk metabolites after intramammary infusion of matrine, a plant alkaloid with anticancer properties, in the form of a chitosan hydrogel into bovine mammary glands with subclinical mastitis. Infusions were continued for seven days, and milk samples were collected on day 1 and day 7 for analysis of the microbiome by 16S rRNA gene sequencing and metabolites by liquid chromatography-mass spectrometry.ResultsMatrine-chitosan hydrogels (MCHs) significantly decreased the somatic cell count on day 7 and the Simpson index indicated that microbial diversity was significantly lower on day 7 than on day 1. On day 7, the numbers of Aerococcus, Corynebacterium_1 and Staphylococcus were significantly lower, while the abundance of Firmicutes was very significantly decreased. The numbers of Probacteria increased, however. In milk samples, we identified 74 differentially expressed metabolites and the MCH infusion group had the most significantly upregulated metabolites including sphingolipids, glycerophospholipids, flavonoids and fatty acyls. Principal component analysis and the orthogonal partial least squares discriminant test confirmed good separation of the milk metabolites. The identification of active milk metabolic pathways after MCH treatment supported the known antimicrobial and anti-inflammatory properties of matrine that are associated with glycerophospholipid metabolism and the sphingolipid metabolic signaling pathways.ConciusionsThese insights into the mechanisms and the corresponding biological responses to matrine demonstrate its potential immunoregulatory activity and emphasize the need for continued investigation.