Ensiling Time and Mixed Microbe Fermented Liquid Modulate In Vitro Digestibility and Rumen Fermentation of Fermented Total Mixed Rations

This study investigated how varying the ensiling period and the level of mixed microbes fermented liquid (MMFL) influences the chemical composition, in vitro degradability, rumen fermentation profile, and microbial ecology of fermented total mixed rations (FTMR). A completely randomized 4 × 4 factorial design was used, incorporating four fermentation durations (0, 7, 14, and 21 days) and four MMFL inclusion rates (0, 0.5, 1.0, and 1.5% of DM). Both factors exerted significant effects on FTMR quality (<i>p</i> < 0.05). The combination of a 14-day fermentation period with 0.5% MMFL consistently produced the most desirable outcomes. Under these conditions, crude protein concentration rose from 12.0% to 14.3% of DM, while neutral-detergent fiber declined from 54.2% to 49.1%. Improvements were also observed in in vitro digestibility, with DM and OM increasing by 9-12% relative to the untreated control (<i>p</i> < 0.05). Fermentation end-products were enhanced, as total volatile fatty acids increased by 15% (92.4 vs. 80.1 mmol/L), and the molar proportion of propionate increased from 24.5 to 29.2 mol/100 mol, thereby lowering the acetate-to-propionate ratio (2.4 vs. 3.0; <i>p</i> < 0.05). Estimated methane production declined by 18% (<i>p</i> < 0.01). Microbial counts reflected a shift toward a more efficient fermentative community, with bacterial and fungal populations increasing by 21% and 18%, and protozoa decreasing by 25% (<i>p</i> < 0.05). Overall, moderate MMFL supplementation during a 14-day ensiling phase enhanced nutrient conservation and fermentation efficiency, suggesting practical value for improving FTMR utilization in ruminant systems. Further in vivo and economic evaluations remain necessary.