Interactive effects of moisture level, lactic acid bacteria, and ensiling duration on fermentation and microbial communities in barnyard millet silage

The present study evaluated the effects of lactic acid bacteria (LAB), moisture content, and ensiling duration on nutrient composition, fermentation quality, and microbial dynamics in barnyard millet (BM) silage. BM forage was ensiled under different moisture conditions using single inoculants ( Lactiplantibacillus plantarum -TC, Lactobacillus brevis 5 M, and Lactobacillus buchneri 6 M) or combined LAB inoculants for 10, 20, and 30 days. LAB inoculation improved fermentation quality by increasing lactic acid and acetic acid production, lowering pH, and inhibiting butyric acid production. Crude protein content remained stable, whereas reductions in neutral detergent fibre (ADF) and acid detergent fibre (NDF) varied with inoculant treatments. Microbial community analysis showed that L. plantarum became the dominant bacterial species, while mixed inoculants promoted greater LAB diversity. Yeast communities were influenced by inoculum, moisture level, and ensiling duration, with Issatchenkia orientalis and Ustilago davisii predominating under different treatments. Fungal communities showed distinct temporal and treatment-dependent patterns, with Ustilago spp. and Mucor circinelloides as major taxa. Correlation analysis revealed positive associations between L. plantarum and lactic acid production, while L. buchneri was associated with acetic acid accumulation. Overall, LAB inoculation was associated with improvements in fermentation quality, microbial stability, and nutrient preservation. However, the inoculants showed significant effects under low-moisture conditions throughout the ensiling period.