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Increased understanding of effects of regulation measures on whole-plant corn silage is important from bacterial community succession, interaction network and predicted functions. According to alpha diversity and meta co-occurrence network, the bacterial communities were more sensitive to storage temperature than LAB inoculants during whole-plant corn ensiling.

Background Bacillus amyloliquefaciens (BA) and Bacillus subtilis (BS) are usually used as feed supplements directly or bacterial inoculants in biological feeds for animals. However, few research have reported the effects of BA and BS on fermentation characteristics and bacterial community successions of whole-plant corn silage during ensiling. If the BA and BS inoculants have positive effects on silages, then they could not only improve fermentation characteristics, but also deliver BA or BS viable cells to ruminants, which would play its probiotic effect. Therefore, the objectives of this study were to investigate the effects of BA and BS on the fermentation, chemical characteristics, bacterial community and their metabolic pathway of whole-plant corn silage. Results Freshly chopped whole-plant corn was inoculated without or with BA and BS, respectively, and ensiled for 1, 3, 7, 14 and 60 d. Results showed that BA and BS inoculations increased lactic acid concentrations of whole-plant corn silages compared with control, and BA inoculation decreased acetic acid concentrations, whereas BS inoculation decreased fiber contents and increased crude protein (CP) content. Higher water-soluble carbohydrate contents and lower starch contents were observed in BA- and BS-inoculated silages compared with that in control. The decreased CP content and increased non-protein nitrogen content were observed in BA-inoculated silage, which was consistent with the higher amino acid metabolism abundances observed in BA-inoculated silage. In addition, it was noteworthy that BA and BS inoculations increased the metabolism of cofactors and vitamins, and decreased the relative abundances of drug resistance: antimicrobial pathways. We also found that the bacterial metabolism pathways were clearly separated into three clusters based on the ensiling times of whole-plant corn silage in the present study. There were no significant differences in bacterial community compositions among the three groups during ensiling. However, BA and BS inoculations decreased the relative abundances of undesirable bacteria such as Acetobacter and Acinetobacter. Conclusion Our findings suggested that the BS strain was more suitable as silage inoculants than the BA strain in whole-plant corn silage in this study.

This study aimed to evaluate the effect of the application of an inoculant and a preservative on the fermentation quality, in vitro digestibility, and aerobic stability of alfalfa silage-based fermented total mixed ration (TMR). The TMR was ensiled with (1) no additives (control), (2) Lactobacillus plantarum (LP), or (3) potassium sorbate (PS). The V-scores of all silages were higher than 80 points during the 30 days of ensiling. The addition of LP and PS had no effects on the in vitro parameters, such as in vitro digestibility and in vitro gas production (p &gt, 0.05). LP-treated silage showed similar fermentation quality and comparable aerobic stability to the control (110 h). The LP only decreased the ammonia nitrogen (NH3-N) content (p &lt, 0.05) during ensiling. The PS significantly increased the pH of TMR silages (p &lt, 0.05). Meanwhile, the addition of PS improved the aerobic stability (&gt, 162 h) of TMR silage, indicated by the higher water-soluble carbohydrate content and lower NH3-N content in comparison with those in the control after aerobic exposure (p &lt, 0.05). The improvement in fermentation quality is extremely small in terms of applying LP in TMR silage based on a large percentage of other silage ingredients. The PS is effective in conserving unpacked TMR silage and showed the potential to reduce the risk of ruminal acidosis in livestock.

Feeding ruminant animals with legume silage, rich in non-protein nitrogen, not only results in serious groundwater pollution but also induces a large amount of methane (CH4) releasing into atmospheric environment. Co-ensiling alfalfa with sainfoin containing condensed tannins in five ratios was investigated on silage fermentation, chemical and bacterial community composition, in vitro ruminal fermentation characteristics, CH4 emission and archaeal and protozoal community. Fresh alfalfa and sainfoin were wilted to dry matter concentrations of 381 and 390 g/kg wet weight, respectively, and chopped to 2–3 cm. Chopped alfalfa and sainfoin were combined in proportions of 1:0 (S0, Control), 0.75:0.25 (S25), 0.5:0.5 (S50), 0.25:0.75 (S75) and 0:1 (S100) on a wet weight basis, respectively. Each treatment was prepared in quadruplicate and fermented for 60 days. After 60 days of fermentation, incorporation of sainfoin into alfalfa before ensiling led to an improvement in silage fermentation and inhibited silage proteolysis in a proportion-dependent manner. Excellent fermentation profiles were observed in S50 and S75 silages, in which the non-protein nitrogen decreased by 11.33% and 13.01%, respectively, compared with S0 silage. As sainfoin proportion increased, the relative abundance of Pediococcus acidilactici linearly decreased (P

Objective To evaluate the effect of lactic acid bacteria and storage temperature on the microbial, chemical and mycotoxin composition of corn silage. Methods Corn was harvested at 32.8% dry matter, and chopped to 1 to 2 cm. The chopped material was subjected to three treatments: i) control (distilled water); ii) 1×106 colony forming units (cfu)/g of Lactobacillus plantarum; iii) 1×106 cfu/g of Pediococcus pentosaceus. Treatments in triplicate were ensiled for 55 d at 20°C, 28°C, and 37°C in 1-L polythene jars following packing to a density of approximately 800 kg/m3 of fresh matter, respectively. At silo opening, microbial populations, fermentation characteristics, nutritive value and mycotoxins of corn silage were determined. Results L. plantarum significantly increased yeast number, water soluble carbohydrates, nitrate and deoxynivalenol content, and significantly decreased the ammonia N value in corn silage compared with the control (p

The effect of Lactobacillus plantarum ‘KR107070’ and a propionic acid-based preservative on the quality and aerobic stability of alfalfa-corn mixed silage ensiled with four ratios was evaluated. A 4 × 4 factorial arrangement was used in a completely randomized design in this study. The chopped alfalfa and corn were made into four alfalfa to corn ratios (containing 20, 40, 60 and 80% corn), each of which was treated with (i) distilled water; (ii) 1 × 106 colony forming units per g of L. plantarum ‘KR107070’; (iii) a chemical additive consisting of 0.24% propionic acid, 0.24% sodium propionate and 0.18% formic acid; (iv) a combination of (ii) and (iii). Treated forages mixture was ensiled with triplicate in a 5-L polythene bucket for 180 days. At silo opening, the fermentation characteristics of silage were enhanced with a lower pH value, higher lactic acid content and lower ammoniacal nitrogen concentration and its relative feed value decreased, with a higher ratio of corn for all treatments. The application of a chemical additive alone, or in combination with L. plantarum ‘KR107070’, decreased ammoniacal nitrogen concentrations and increased water soluble carbohydrates contents than control or L. plantarum ‘KR107070’ for each silage of four ratios, in addition to enhancing the aerobic stability of 60 and 80% corn silages compared to L. plantarum ‘KR107070’. The 60% corn silage, treated with combining 0.24% propionic acid, 0.24% sodium propionate and 0.18% formic acid with L. plantarum ‘KR107070’, is better in terms of conservation quality and aerobic stability.

Simple Summary Alfalfa (Medicago sativa) is commonly used as a high-quality protein source in fermented total mixed ration (FTMR) for ruminants. This study evaluated the fermentation characteristics, chemical compositions, and protein fractions of FTMR using fresh-alfalfa as the main ingredients replacing ensiled-alfalfa. The results showed that fresh-alfalfa FTMR exhibited a similar pH, propionic acid content and neutral detergent fiber, nonprotein, and variable to slow protein and indigestible protein levels in comparison to ensiled-alfalfa FTMR. Therefore, the use of fresh-alfalfa as a main ingredient in FTMR is promising. Abstract Alfalfa (Medicago sativa) is one of the high protein ingredients of fermented total mixed ration (FTMR). Additionally, FTMR is widely used to satisfy the nutrition requirements of animals. This study was conducted to confirm the fermentation characteristics, chemical compositions and protein fractions changes when replacing ensiled-alfalfa with fresh-alfalfa in FTMR with additives. Three additives were separately applied to fresh-alfalfa total mixed ration (TMR) and ensiled-alfalfa TMR, including molasses (MOL), Lactobacillus plantarum (LP) and MOL plus LP (MOL+LP). The same volume of distilled water was sprayed onto the prepared TMR as performed for the control (CK). Each treatment included 18 repetitions and opened 3 repetitions at each fermenting day (1, 3, 7, 15, 30 and 60 d). The results showed that fresh-alfalfa FTMR (F-FTMR) exhibited slight changes in the fermentation characteristics during the first 7 d and showed similar trends in terms of the pH and organic acids content to ensiled-alfalfa FTMR (E-FTMR). The lactic acid contents of F-FTMR were significantly lower than those of E-FTMR at 60 d fermentation and the ammonia nitrogen contents were lower than E-FTMR during the entire fermenting period. The crude protein of the F-FTMR was enhanced after 60 d of fermenting. F-FTMR supplemented with MOL+LP exhibited a lower nonprotein nitrogen content, variable to slow protein and indigestible protein contents, and higher fast degradable protein and true protein degraded intermediately contents at 60 d fermenting, indicating that it effectively inhibited protein degradation.

Silage fermentation improves when alfalfa is ensiled along with whole-plant corn, yet so far the impact of their mixing ratios on individual bacteria during fermentation is weakly described. The objective of this work was to explore the effect of combining alfalfa with whole-plant corn in different ratios on silage bacterial community and fermentation profile over the entire ensiling process. The alfalfa at the early bloom stage and whole-plant corn at around the 1/3 milk line stage were freshly chopped and blended at proportions of 1:0 (served as Control), 0.8:0.2 (Mix 1), 0.6:0.4 (Mix 2) and 0:1 on a fresh matter basis. Each treatment was prepared in triplicate and ensiled in plastic bag silos for 0, 3, 7, 14 and 35 days. The fermentation coefficient of fresh matter used to asses a forage ensilability linearly increased, with more corn in the mixture. During the first 3 days of fermentation, the greatest pH drop occurred in all silages. Seven days later, the silage pH in Mix 1, Mix 2 and corn further decreased, and alfalfa silage pH slightly increased, however. After 35 days of ensiling, alfalfa silage was poorly fermented, shown by a high level of pH (4.69), acetic acid (35.7 g/kg dry matter), propionic acid (29.0 g/kg dry matter), n-butyric acid (12.1 g/kg dry matter) and ammonia nitrogen (108 g/kg total nitrogen). Silage fermentation was significantly improved with corn percentage increasing from 20 % to 100 %. In the early stage of fermentation, Lactobacillus and Pediococcus relative abundance noticeably increased in alfalfa, Mix 1 and Mix 2 silages, whereas Leuconostoc abundance was noticeably enriched and outnumbered Lactobacillus and Pediococcus in corn silage. During 7–35 days of ensiling, the bacterial community structure in all silages comparatively stayed stable. Besides, Lactobacillus abundance increased while Pediococcus and Weissella richness decreased with more corn, at each period above. The bacterial community composition of all fresh materials was similar at the genus level, consisting mainly of Weissella, Leuconostoc, Pantoea, Enterobacter, Pseudomonas and Rosenbergiella. However, the development of Lactobacillus, Pediococcus and Leuconostoc species in alfalfa, Mix 1 and Mix 2 silages in the early stage of fermentation was different from corn silage. Subsequently, the bacterial community structure in all silages relatively stayed stable. Overall silage fermentation quality was better when alfalfa was ensiled in a mixture with at least 20 % whole-plant corn on a fresh matter basis.

This study assessed the effects of antibacterial peptide-producing Bacillus subtilis (BS), Lactobacillus buchneri (LB), or their combination on fermentation, proteolysis, aerobic stability, and microbial communities during ensiling and aerobic exposure phases of alfalfa silage. The results showed that the BS-treated silage displayed a higher lactic acid concentration, less proteolysis, and higher aerobic stability than those in the control silage. Both LB and BS treatments increased Lactobacillus and Ascochyta abundance, and decreased Enterococcus and Sporormiacea abundance after 60 d of fermentation. LB and BS also inhibited the growth of Enterococcus after 3 d of aerobic exposure but similar to the control silage, the fungal community of BS silage was dominated by Candida and Pichia after 9 d of aerobic exposure. Therefore, inoculation of BS improved silage fermentation quality, aerobic stability and bacterial community during ensiling and after 3 d of aerobic exposure.

The effect of Lactobacillus buchneri, alone or in combination with Lactobacillus plantarum, on the fermentation characteristics and aerobic stability of whipgrass silage was investigated under laboratory conditions. The inoculants were applied at 2 × 106 colony forming units g−1. Uninoculated silages served as control. After treatment, the chopped materials were ensiled in 1.5-L anaerobic glass jars. Three jars per treatment were sampled on days 2, 4, 8, 15, 30, 45 and 90. After 90 days of fermentation, the silages were subjected to an aerobic stability test lasting 5 days, in which CO2 production, change in pH and microbial numbers were measured to determine the extent of aerobic deterioration. At the end of the fermentation period 90 days, inoculation with L. buchneri or in combination with L. plantarum significantly resulted in higher levels of acetic acid in the silages than other treatments (P