Alfalfa silage production is challenged by high moisture, strong buffering capacity, and low water-soluble carbohydrate concentrations, yet long-term storage is common under Korean production conditions. This study evaluated how target moisture content at ensiling and a lactic acid bacterial inoculant affect the chemical composition, fermentation profile, and bacterial community of alfalfa silage stored for 12, 18, or 24 months. Alfalfa harvested at National Institute of Animal Science (NIAS, Cheonan) was ensiled at target moisture levels of 30, 40, 50, 60, or 70% with either no additive (CON) or a mixed inoculant containing Lactococcus lactis and Pediococcus pentosaceus (INO). Moisture content exerted the dominant and most consistent influence on silage quality. Crude protein generally decreased and acid detergent fiber (ADF) generally increased as moisture content increased, whereas neutral detergent fiber (NDF) responses were not consistently quadratic across all storage periods. The 40–50% moisture treatments showed the most favorable lactic fermentation, with lower pH and higher lactic acid concentrations than the drier 30% and wetter 60∽70% treatments. Inoculation effects were limited and depended on moisture level and storage duration; significant inoculation-related effects were observed for pH and propionic acid at 12 months and lactic acid at 24 months, but inoculation did not consistently improve lactic acid accumulation. The 60∽70% moisture treatments showed elevated butyric acid and Clostridium relative abundance, indicating increased risk of undesirable fermentation. At 70% moisture, Clostridium relative abundance was numerically lower in INO than in CON; however, this pattern was not consistently supported by butyric acid responses. Overall, controlling ensiling moisture to approximately 40∽50% appears more reliable than relying on inoculation alone for maintaining long-term alfalfa silage quality.

ABSTRACT
Ⅰ. INTRODUCTION
Ⅱ. MATERIAL AND METHODS
    1. Plant material and site
    2. Treatments, sampling, and silage preparation
    3. Sample processing and chemical analyses
    4. Fermentation characteristics
    5. DNA extraction and 16S rRNA gene sequencing
    6. Statistical analysis
Ⅲ. RESULTS AND DISCUSSION
    1. Chemical composition
    2. Fermentation characteristics
    3. Microbial community
Ⅳ. CONCLUSION
Ⅴ. ACKNOWLEDGEMENT
Ⅵ. REFERENCES

• Seung Min Jeong(Forages Production Systems Division, National Institute of Animal Science, Cheonan 31000, Republic of Korea)
• Ki Won Lee(Forages Production Systems Division, National Institute of Animal Science, Cheonan 31000, Republic of Korea)
• Hyung Soo Park(Forages Production Systems Division, National Institute of Animal Science, Cheonan 31000, Republic of Korea) Corresponding author