The objective of this study was to evaluate the effects of Lactiplantibacillus plantarum or cellulase on the fermentation characteristics and bacterial community of mixed alfalfa (Medicago sativa L., AF) and Leymus chinensis (LC) silage. The harvested alfalfa and Leymus chinensis were cut into 1–2 cm lengths by a crop chopper and they were thoroughly mixed at a ratio of 3/2 (wet weight). The mixtures were treated with no addition (CON), Lactiplantibacillus plantarum (LP, 1 × 10⁶ cfu/g fresh material), cellulase (CE, 7.5 × 10² U/kg fresh material) and their combination (LPCE). The forages were packed into triplicate vacuum-sealed, polyethylene bags per treatment and ensiled for 1, 3, 5, 7 and 30 d at room temperature (17–25 °C). Compared to the CON groups, all the additives increased the lactic acid content and decreased the pH and ammonia nitrogen content over the ensiling period. In comparison to the other groups, higher water-soluble carbohydrate contents were discovered in the CE-inoculated silages. Compared to the CON groups, the treatment with LPCE retained the crude protein content and reduced the acid detergent fiber content. The principal coordinate analysis based on the unweighted UniFrac distance showed that individuals in the AF, LC, CON and LPCE treatment could be significantly separated from each other. At the genus level, the bacterial community in the mixed silage involves a shift from Cyanobacteria_unclassified to Lactobacillus. Lactobacillus dominated in all the treatments until the end of the silage, but when added with Lactiplantibacillus plantarum, it was more effective in inhibiting undesirable microorganisms, such as Enterobacter, while reducing microbial diversity. By changing the bacterial community structure after applying Lactiplantibacillus plantarum and cellulase, the mixed silages quality could be further improved. During ensiling, the metabolism of the nucleotide and carbohydrate were enhanced whereas the metabolism of the amino acid, energy, cofactors and vitamins were hindered. In conclusion, the relative abundance of Lactobacillus in the mixed silage increased with the addition of Lactiplantibacillus plantarum and cellulase, which also improved the fermentation quality.

Most oat forage has low water-soluble carbohydrates (WSC), which may be the main limited factor for silage fermentation safely, but oat is rich in cellulose and hemicellulose; therefore, we assume that xylanase and cellulase as additives can reduce the content of cellulose and xylan in oat silage, increase the microbial fermentable sugar content, and improve the fermentation quality of the silage. After wilting, oats were treated as follows: (i) distributed water (CK); (ii) silages inoculated with xylanase (X); (iii) silages inoculated with cellulase (C), ensiling for 3 days (early stage of silage) and 60 days (late stage of silage), respectively, after ensiling 60 days for a 5-day aerobic exposure study. The pH, neutral detergent fiber (NDF), and acid detergent fiber (ADF) were significantly reduced by xylanase and cellulase treatment during the late stage of silage, and the concentration of lactic acid, acetic acid, and ammonia nitrogen increased remarkably. The WSC content reached its peak with xylanase treatment during the late stage of silage. The content of crude protein (CP) was not affected by additives but by the silage period; CP and ether extract (EE) significantly increased during the late stage of silage compared to the early stage. After ensiling, the bacterial community showed that xylanase and cellulase treatment increased the relative abundance of lactic acid bacteria. Lactobacillus has a higher relative abundance with cellulase treatment after 60 days of ensiling; this can effectively reduce the pH of silage and ensure long-term, stable storage of silage. Cellulase and xylanase increased bacterial diversity during aerobic exposure and improved the aerobic stability of silage significantly. This study indicated that different additives and silage periods had significant effects on chemical compositions, fermentation quality, and bacterial community; meanwhile, both additives improved the aerobic stability of silage. In summary, when the WSC of oat is low, cellulase and xylanase have good effects as silage additives, and the comprehensive effect of cellulase is more prominent.

Based on the low content of water-soluble carbohydrate (WSC) and lactic acid bacteria (LAB) attachment in oat raw materials, we assumed that the neutral detergent fiber (NDF) content of oat can be reduced by adding cellulase or xylanase. The concentration of metabolizable sugars will be increased, which will assist the oat’s bacterial community in fermentation and obtain a better quality of oat silage. After wilting the oat, it was treated as follows: (1) distributed water (CK); (2) silages inoculated with xylanase (X); and (3) silages inoculated with cellulase (C), ensiling for 3, 7, 14, 30, and 60 days. Cellulase and xylanase treatments both alter the fermentation and nutritional quality of ensiled oat, resulting in lower NDF, acid detergent fiber (ADF), cellulose, and hemicellulose contents, increased lactic acid and acetic acid contents, and a significant decrease in ensiling environment pH. The bacterial community undergoes significant changes with cellulase and xylanase treatments, with a significant increase in Lactobacillus abundance in the C_14, X_30, C_30, X_60, and C_60 treatment groups, while Weissella abundance gradually decreases with longer ensiling times. Two exogenous fibrolytic enzymes also alter the bacterial diversity of ensiled oat, with different bacterial species and abundances observed in different treatment groups. Ensiled oat treated with cellulase and xylanase experiences significant changes in its own bacterial community, particularly in the abundance of Lactobacillus. These changes result in improved fermentation and nutritional quality of oat, but the higher metabolism levels observed after 60 days of ensiling with cellulase treatment may lead to energy loss.

This study investigated whether two exopolysaccharides could serve as exogenous carbon sources to enhance fermentation quality in oat silage, providing a theoretical foundation for their future application in silage. The oats were harvested at the heading stage and, following a period of wilting, were chopped into 2–3 cm lengths for the ensiling experiment. The treatments applied were as follows: (1) a control group (CK), which received only sterile water; (2) a group with added dextran (D); and (3) a group with added levan (L). The fermentation process was monitored at various intervals: 3, 7, 14, 30 and 60 days (d), respectively. Following 60 days of ensiling, the silage was subjected to a 5‐day period of aerobic exposure (AE). EPS changed the fermentation quality of silage, altered the composition of the bacterial community, and had an impact on the feature dissimilarity between sample groups. Meanwhile, EPS showed different regulatory effects on carbohydrate metabolism at different fermentation times. EPS treatment increased the lactic acid content and decreased the pH of silage. After 60 days of fermentation, the treatment also increased the relative abundance of Lactobacillus . Dextran and levan increased the relative abundance of Hafnia–Obesumbacterium and Sediminibacterium , respectively. Under the treatment of dextran, silage retained more WSC content and achieved higher aerobic stability. Upon comparing the bacterial correlation networks, it became evident that the fermentation time altered the composition of inter‐bacterial correlations. In conclusion, EPS can effectively enhance the fermentation quality of oat silage, with dextran yielding the most pronounced positive effects.