The objective of this study was to select lactic acid bacteria (LAB) isolated from the rumen fluid and feces of dairy cows, and evaluate their effects on silage quality of alfalfa after 30 or 60 days of ensiling. One hundred and four LAB strains were isolated from rumen fluid and feces of six dairy cows, of which four strains (Lactobacillus plantarum F1, L. plantarum F50, Lactobacillus salivarius L100, and Lactobacillus fermentum L120) and one commercial inoculant (GFG) isolated from forage were employed for further study. The silages treated with F1 had the lowest (P < 0.05) pH value and the highest (P < 0.05) lactic acid (LA) content in all treatments. Besides, higher (P < 0.05) in vitro digestibility was also observed in F1-treated silage after 60 days of ensiling. The microbial analysis showed that the Lactobacillus abundance in the F1-treated silages increased to 60.32%, higher than other treatments (5.12–47.64%). Our research indicated that strain F1 could be an alternative silage inoculant, and dairy cows could be a source for obtaining excellent LAB for ensiling.

Silage quality remains an important issue in farming, as do limitations in the range of products suitable for animal fodder. We therefore explored the microorganisms that are critical for the fermentation quality of paper mulberry silage. Low (unwilted) and high (wilted) dry matter (DM) paper mulberry were harvested at two cutting times. These were ensiled for 0, 3, 7, 14, and 56 days, respectively. Compared with unwilted silages, wilting significantly decreased (p < 0.05) silage pH value, ammonia‐N concentration, and yeast counts but increased (p < 0.05) lactic acid content. In addition, higher (p < 0.05) crude protein (CP) contents were also observed in wilted silages. Next‐generation sequencing (NGS) analysis revealed that wilting reduced the abundance of Enterobacter, while increasing that of Lactobacillus. Single‐molecule real‐time sequencing (SMRT) revealed that the silage was enriched in the lactic acid bacteria (LAB), Lactobacillus rhamnosus after wilting, which showed a positive correlation with CP and lactic acid content. We conclude that wilting may help preserve paper mulberry silage, facilitating its use as a new fodder resource. Moreover, L. rhamnosus has the potential to be developed as a new inoculant for the modulation in wilted silages, particularly paper mulberry silage.

The objective of this study was to investigate how storage temperatures influence the bacterial community of oat silage during the ensiling process via PacBio single molecule, real-time sequencing technology (SMRT). Forage oat was ensiled at four different temperatures (5 °C, 10 °C, 15 °C, and 25 °C) and ensiling days (7, 14, 30, and 60 days). With the rise in storage temperature, the lactic acid content showed an increased trend. Acetic acid production was observed highest in silage fermented at 5 °C compared with other treatments, and Enterococcus mundtii was also the dominant bacterial species. Lactiplantibacillus pentosus and Loigolactobacillus rennini were exclusively detected in silages at 10 °C, 15 °C, and 25 °C, and dominated the fermentation after 60 days of ensiling at 10 °C and 25 °C, respectively. In addition, L. pentosus, L. rennini, and E. mundtii may be related to changes in the fermentation products due to the differences in ensiling temperature. In conclusion, results of this study improve our understanding of the complicated microbial composition underlying silage fermentation at low temperatures, which might contribute to target-based regulation methods for enhancing silage quality and developing new inoculants.

The objective of this study was to investigate the effects of wilting and additives on the fermentation quality, structural and non-structural carbohydrate composition of mulberry silages. The selected LAB strains Lactobacillus (L.) plantarum 'LC279063' (L1), commercial inoculant Gaofuji (GF), and Trichoderma viride cellulase (CE) were used as additives for silage preparation. Silage treatments were designed as control (CK), L1, GF, or CE under three wilting rates, that is wilting for 0, 2 or 4 hours (h). After ensiling for 30 days, the silages were analyzed for the chemical and fermentation characteristics. The results showed that wilting had superior effects on increasing the non-structural carbohydrate concentration and degrading the structural carbohydrate. After ensiling for 30 days, L1 generally had a higher fermentation quality than other treatments, indicated by the lower pH value, acetic acid, propionic acid and ammonia nitrogen (NH3-N) content, and the higher lactic acid, water soluble carbohydrate (WSC), glucose, galactose, sucrose and cellobiose concentration (p<0.05) at any wilting rate. Wilting could increase the ratio of lactic acid/acetic acid and decrease the content of NH3-N. The results confirmed that wilting degraded the structural carbohydrate and increased the non-structural carbohydrate; and L1 exhibited better properties in improving fermentation quality and maintaining a high non-structural carbohydrates composition compared with the other treatments.

There are few studies on the application of lactic acid bacteria in the reduction of anti-nutrient factors in paper mulberry silage. This study aimed to investigate the effects of different lactic acid bacteria on the fermentation quality and the amount of anti-nutritional factors in paper mulberry silage. Two strains of Lactobacillus plantarum (GX, isolated from paper mulberry silage; GZ, provided by Sichuan Gaofuji Biotechnology Co. Ltd.) were added as silage additives. On days 7, 15, 30 and 60 of the ensiling process, the fermentation quality, and the amount of anti-nutritional factors were measured. Compared with the control group, inoculation with Lactobacillus plantarum could rapidly reduce pH values, leading to lower NH₃-N/TN. Besides, it also significantly increased the lactic acid content (p < 0.05). The two strains of L. plantarum significantly reduced the content of hydrolysed tannin, condensed tannin, total tannin, oxalic acid, phytic acid and saponin (p < 0.05). Overall, this study found that the addition of lactic acid bacteria could significantly improve the fermentation quality of paper mulberry and reduce the amount of anti-nutrient factors (p < 0.05).

The naturally attached phyllosphere microbiota play a crucial role in plant-derived fermentation, but the structure and function of phyllosphere endophytes remain largely unidentified. Here, we reveal the diversity, specificity, and functionality of phyllosphere endophytes in alfalfa (Medicago sativa L.) through combining typical microbial culture, high-throughput sequencing, and genomic comparative analysis. In comparison to phyllosphere bacteria (PB), the fermentation of alfalfa solely with endophytes (EN) enhances the fermentation characteristics, primarily due to the dominance of specific lactic acid bacteria (LAB) such as Lactiplantibacillus, Weissella, and Pediococcus. The inoculant with selected endophytic LAB strains also enhances the fermentation quality compared to epiphytic LAB treatment. Especially, one key endophytic LAB named Pediococcus pentosaceus EN5 shows enrichment of genes related to the mannose phosphotransferase system (Man-PTS) and carbohydrate-metabolizing enzymes and higher utilization of carbohydrates. Representing phyllosphere, endophytic LAB shows great potential of promoting ensiling and provides a novel direction for developing microbial inoculant.