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This study evaluated the effect of methionine on in vitro methane (CH 4 ) production, rumen fermentation, amino acid (AA) metabolism, and rumen microbiota in a low protein diet. We evaluated three levels of methionine (M0, 0%; M1, 0.28%; and M2, 1.12%) of in the presence of sodium nitrate (1%) in a diet containing elephant grass (90%) and concentrate (10%). We used an in vitro batch culture technique by using rumen fluid from cannulated buffaloes. Total gas and CH 4 production were measured in each fermentation bottle at 3, 6, 9, 12, 24, 48, 72 h of incubation. Results revealed that M0 decreased ( p &lt; 0.001) the total gas and CH 4 production, but methionine exhibited no effect on these parameters. M0 decreased ( p &lt; 0.05) the individual and total volatile fatty acids (VFAs), while increasing ( p &lt; 0.05) the ruminal pH, acetate to propionate ratio, and microbial protein content. Methionine did not affect ruminal AA contents except asparagine, which substantially increased ( p = 0.003). M2 increased the protozoa counts, but both M0 and M1 decreased ( p &lt; 0.05) the relative abundance of Firmicutes while increasing ( p &lt; 0.05) the Campilobacterota and Proteobacteria. However, Prevotella and γ-Proteobacteria were identified as biomarkers in the nitrate group. Our findings indicate that methionine can increase ruminal asparagine content and the population of Compylobactor .

Effect of Methionine Supplementation on Rumen Microbiota, Fermentation, and Amino Acid Metabolism in In Vitro Cultures Containing Nitrate