Milk fat depression (MFD) syndrome represents a significant drawback to the dairy industry. The aim of this study was to unravel the ruminal metabolome-microbiome interaction in response to diet-induced MFD in dairy cows. Twelve healthy second parity Holstein dairy cows (days in milk (DIM) = 119 ± 14) were randomly assigned into control (CON, n = 6) group and treatment (TR, n = 6) group. Cows in TR group received a high-starch total mixed ration (TMR) designed to induce an MFD syndrome. Decreased milk fat yield and concentration in TR cows displayed the successful development of MFD syndrome. TR diet increased the relative abundance of Prevotella and decreased the relative abundance of unclassified Lachnospiraceae, Oribacterium, unclassified Veillonellaceae and Pseudobutyrivibrio in ruminal fluid. Metabolomics analysis revealed that the ruminal fluid content of glucose, amino acids and amines were significantly increased in TR cows compared with CON cows. Correlation analysis revealed that the concentration of amines and amino acids were highly correlated with the abundance of Oribacterium, Pseudobutyrivibrio, RC9_gut_group, unclassified BS11_gut_group and Selenomonas. In general, these findings revealed that TR diet reduced the rumination time and altered rumen fermentation type, which led to changes in the composition of ruminal microbiota and metabolites, and caused MFD.

The production and consumption of pineapple creates large quantities of residues. Ensiling these residues might help to minimize the waste burden and meet the intensive feed demand for ruminants. Proper lactic acid bacteria (LAB) are not only responsible for pineapple residual silage fermentation, but might also deliver probiotics. The aim of this study was to isolate LAB strains with probiotic functions, and to enhance intestinal antioxidant capacity from naturally fermented pineapple residues. A total of 47 LAB isolates with gram-positive, catalase-negative, nonhemolytic properties were used for probiotic screening. Lactiplantibacillus plantarum (L. plantarum) A1, C1 and C10 were susceptible to rifampicin, gentamicin and erythromycin, did not contain virulence factor-coding genes and showed good tolerance to acid (pH 3.0), 0.5% bile salt and simulated gastric and intestinal fluid. Their hydrophobicity indices were 71.92%, 45.50% and 66.90%, respectively. All of them were able to adhere to bovine jejunum epithelial cells (BJECs) and to antagonize Escherichia coli F5 and Salmonella Dublin. These three LAB strains tolerated hydrogen peroxide and significantly decreased (p < 0.05) reactive oxygen species levels in BJECs. In addition, L. plantarum C1 and C10 significantly increased (p < 0.05) the total antioxidant capacity in BJECs in the presence of 200 μmol/L hydrogen peroxide condition. L. plantarum A1, C1 and C10 are potential probiotics isolated from pineapple residual silage. This study aims to promote pineapple residue’s utilization in the feed industry.