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.

Silage of native grasses can alleviate seasonal forage supply imbalance in pastures and provide additional sources to meet forage demand. The study aimed to investigate the effects of Lactobacillus plantarum (LP), Lactobacillus buchneri (LB), and Lactobacillus plantarum in combination with Lactobacillus buchneri (PB) on the nutritional quality, fermentation quality, and microbial community of native grass silage at 2, 7, 15, and 60 days after ensiling and at 4 and 8 days after aerobic exposure. The results showed that dry matter content, crude protein content, the number of lactic acid bacteria, and lactic acid and acetic acid content increased and pH and ammonia nitrogen content decreased after lactic acid bacteria (LAB) inoculation compared with the control group (CK). LP had the lowest pH and highest lactic acid content but did not have greater aerobic stability. LB maintained a lower pH level and acetic acid remained at a higher level after aerobic exposure; aerobic bacteria, coliform bacteria, yeast, and molds all decreased in number, which effectively improved aerobic stability. The effect of the compound addition of LAB was in between the two other treatments, having higher crude protein content, lactic acid and acetic acid content, lower pH, and ammonia nitrogen content. At the phylum level, the dominant phylum changed from Proteobacteria to Firmicutes after ensiling, and at the genus level, Lactiplantibacillus and Lentilactobacillus were the dominant genera in both LAB added groups, while Limosilactobacillus was the dominant genus in the CK treatment. In conclusion, the addition of LAB can improve native grass silage quality by changing bacterial community structure. LP is beneficial to improve the fermentation quality in the ensiling stage, LB is beneficial to inhibit silage deterioration in the aerobic exposure stage, and compound LAB addition is more beneficial to be applied in native grass silage.

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.

During aerobic exposure of silage, the fatty acid and amino acid composition may alter the quality and palatability, resulting in economic losses in livestock production. The objective of this study was to evaluate the effects of Lactiplantibacillus plantarum (LP), Lenti Lentilactobacillus buchneri (LB), and a mixture of LP and LB (PB) on the fatty acids, amino acids, and antioxidant capacity of Leymus chinensis silage during aerobic exposure. The lactic acid bacteria were added at 1 × 10⁶ CFU/g. The silage treatments were opened after 60 days of fermentation, and sampled on days 0, 4, and 8 of aerobic exposure. The LB group had higher total fatty acid and polyunsaturated fatty acid content, and less decrease in amino acid content and antioxidant capacity, while the LP group had a higher monounsaturated fatty acid content but a larger decrease in all indicators after exposure. Correlation analysis showed that Lactobacillus, Cryptococcus, Penicillium, and Thermoascus were more correlated with fatty acid changes, and that Lactobacillus, Actinomyces, Clostridium, and Penicillium were more correlated with amino acid changes. In conclusion, Lentilactobacillus buchneri could effectively improve the antioxidant capacity and fatty acid and amino acid contents of Leymus chinensis silage during aerobic exposure, while Lactiplantibacillus plantarum could effectively improve the content of each index of Leymus chinensis silage at opening, but deterioration was faster during aerobic exposure.

Nitrites are universally acknowledged natural toxic substances that frequently lead to poisoning in humans and animals. During fermentation, certain microorganisms utilize a portion of the nitrogen element and reduce nitrates to nitrites through specific metabolic pathways. In this study, a highly effective lactic acid bacterial strain, Lactiplantibacillus plantarum A50, was isolated and screened from alfalfa silage for its remarkable ability to degrade nitrites. L. plantarum A50 exhibits exceptional nitrite removal capacity, with a degradation rate of 99.06% within 24 h. Furthermore, L. plantarum A50 demonstrates normal growth under pH values ranging from 4 to 9 and salt concentrations of 5%, displaying excellent tolerance to acidity, alkalinity, and salinity. Additionally, it undergoes fermentation using various carbon sources. Within the first 6–12 h of culture, L. plantarum A50 primarily achieves nitrite degradation through non-acidic processes, resulting in a degradation rate of 82.67% by the 12th hour. Moreover, the metabolites produced by L. plantarum A50 exhibit a synergistic interaction with acidity, leading to a nitrite degradation rate of 98.48% within 24 h. Notably, both L. plantarum A50 and MRS broth were found to degrade nitrites. Consequently, a non-targeted metabolomic analysis using LC-MS was conducted to identify 342 significantly different metabolites between L. plantarum A50 and MRS broth. Among these, lipids and lipid-like molecules, organic acids and derivatives, organic oxygen compounds, and organoheterocyclic compounds emerged as the main constituents. Lipids and lipid-like molecules, derivatives of glucose and galactose, amino acids and their derivatives, as well as organoheterocyclic compounds, are likely to play a role in nitrite elimination. Through the enrichment analysis of differential metabolic pathways using KEGG, nine distinct pathways were identified. These pathways provide essential nutrients, maintain cellular structure and function, participate in substance transport, regulate metabolic activities, and enhance resistance against pathogenic microorganisms in L. plantarum A50.

In this study, the triterpenoids in the leaves of Lactuca indica L.cv. Mengzao (LIM) were extracted via microwave-assisted ethanol extraction, and the optimum extraction conditions for triterpenoids were determined through single-factor experiments and the Box–Behnken method. The effects of three factors (solid–liquid ratio, microwave power and extraction time) on the total triterpenoids content (TTC) were evaluated. The TTC of different parts (roots, stems, leaves and flowers) of LIM in different growth stages was studied, and the scavenging effects of the highest TTC parts on DPPH, ABTS and hydroxyl free radicals were investigated. The results showed that the optimum extraction conditions for microwave-assisted extraction of total triterpenoids from LIM leaves were as follows: solid–liquid ratio of 1:20 g/mL; microwave power of 400 W; and extraction time of 60 min. Under these conditions, the TTC was 29.17 mg/g. Compared with the fresh raw materials, the TTC of the materials increased after freeze drying. The leaves of LIM had the highest TTC, and the flowering stage was the best time. The triterpenoids from the leaves had a strong ability to eliminate DPPH and ABTS free radicals, and the elimination effect of dried leaves was better than that of fresh leaves, while the elimination effect of hydroxyl free radicals was not obvious. The tested method was used to extract total triterpenoids from LIM using a simple process at low cost, which provides a reference for developing intensive processing methods for L. indica.