biostudies-literatureFioriniello SItalian Ministry for Economic DevelopmentEuropean Union NextGenerationEU8074https://www.ebi.ac.uk/biostudies/studies/S-EPMC11890600biostudies-literatureUnknown15(1)Nutrition impacts the epigenetic signature, including DNA methylation. The aim of this study was to identify genomic regions differentially methylated in the rumen of Italian Mediterranean dairy buffaloes fed green forage [Total Mixed Ration (TMR) + ryegrass green feed (30% of diet)] compared to those receiving a standard TMR diet, through Reduced Representation Bisulfite Sequencing. We found 6571 differentially methylated genomic regions (DMRs), 51.73% hypomethylated and 48.27% hypermethylated. DMRs were uniformly dispersed in genes and intergenic regions and along chromosomes. Genes-associated DMRs were mainly hypomethylated, while intergenic DMRs were mostly hypermethylated. We highlighted 4648 genes associated with DMRs (differentially methylated genes, DMGs), mostly protein-coding genScientific reportsGreen forage impacts on the DNA methylation in the ruminal wall of Italian mediterranean dairy buffaloes.PMC11890600CN00000022F/200016/01-03/X45Fioriniello SDella Ragione FBifulco GCampanile GSalzano AAiese Cigliano RfalseGreen forage impacts on the DNA methylation in the ruminal wall of Italian mediterranean dairy buffaloes.Nutrition impacts the epigenetic signature, including DNA methylation. The aim of this study was to identify genomic regions differentially methylated in the rumen of Italian Mediterranean dairy buffaloes fed green forage [Total Mixed Ration (TMR) + ryegrass green feed (30% of diet)] compared to those receiving a standard TMR diet, through Reduced Representation Bisulfite Sequencing. We found 6571 differentially methylated genomic regions (DMRs), 51.73% hypomethylated and 48.27% hypermethylated. DMRs were uniformly dispersed in genes and intergenic regions and along chromosomes. Genes-associated DMRs were mainly hypomethylated, while intergenic DMRs were mostly hypermethylated. We highlighted 4648 genes associated with DMRs (differentially methylated genes, DMGs), mostly protein-coding gen2025-01-01T00:00:00Z2025 Mar2025-04-04T08:22:33.358Z2025-04-04T08:22:33.358ZS-EPMC118906004005754210.1038/s41598-025-91969-y