Abstract
Native grasses possess rich diversity and contribute to enhancing the nutritional value of silage, promoting digestion and absorption, thus improving the health of livestock such as cattle and sheep. However, in northern China, the silage fermentation process occurs at relatively low temperatures, necessitating the use of cold-tolerant lactic acid bacteria (LAB). This study examined the effect of Pediococcus acidilactici (L10), a strain selected for its low-temperature tolerance, added to native grass silage at 5°C (LT), 15°C (MT), and room temperature 25°C (CK) for 60 days. The organization of the microbial community and the metabolomic profiles were examined. The results showed that temperature significantly (P < 0.05) influenced the pH, lactic acid (LA) concentration, and LAB populations of the silage after 60 days. The water-soluble carbohydrates (WSC) and crude protein (CP) contents in the LT treatment were significantly higher than those in the CK treatment, and the pH in the LT treatment was significantly lower than in the CK treatment. In terms of the dynamic alterations within the microbial community, Pediococcus acidilactici prevailed in the LT treatment, whereas Lactobacillus plantarum was the major genus in the MT treatment, and the CK treatment was characterized by the dominance of Lactobacillus plantarum and Levilactobacillus brevis. The study also revealed that bacterial behavior and metabolism were influenced by two-component systems and quorum sensing. At 5°C the upregulation of citric acid, salicylic acid, and L-proline was ascribed to the modification of glycolysis and the tricarboxylic acid cycle. Salicylic acid was significantly (P < 0.05) positively correlated with Lactiplantibacillus plantarum, while L-proline had significantly (P < 0.05) positive correlations with Pediococcus acidilactici, Lactococcus lactis, and Weissella confusa. These findings suggest that the addition of isolated Pediococcus acidilactici can enhance the quality of low-temperature native grass silage by regulating microbial metabolic pathways and community composition.IMPORTANCEThis study aimed to screen and identify low-temperature-resistant lactic acid bacteria (LAB) strains from native fermented silage of grassland pastures, evaluating their impact on silage quality in cold conditions. Under natural conditions, LAB on forage grasses are present in low numbers and exhibit insufficient activity, which is further hindered by low temperatures during ensiling, leading to slow fermentation. The findings highlighted the effects of low temperatures on the microbial community, fermentation characteristics, and metabolomic profiles of silage. After anaerobic fermentation, the main LAB strains at different temperatures were Levilactobacillus brevis, Lactiplantibacillus plantarum, and Pediococcus acidilactici, with Pediococcus acidilactici being dominant at 5°C. Temperature significantly affected the pH, lactic acid content, and water-soluble carbohydrates of silage, indicating an interaction between LAB strains and fermentation temperature. The study suggests that adding Pediococcus acidilactici can enhance silage quality by regulating microbial metabolic pathways and composition under low-temperature conditions.