Effects of Condensed Tannins on Bacterial and Fungal Communities during Aerobic Exposure of Sainfoin Silage.

BACKGROUND: Sainfoin is a forage legume that is widely distributed around the world and is beneficial for animals owing to the characteristics of its condensed tannins (CTs), which, from certain plants, can prolong the aerobic stability of silage. METHODS: The present study investigated whether sainfoin CTs can prolong aerobic stability by adding polyethylene glycol (PEG) to inactivate CT activity in the silage system. RESULTS: The results showed that aerobic stability increased under the PEG treatment (p < 0.05). Ammonia nitrogen (0.71 g/kg DM vs. 0.94 g/kg DM; p < 0.05) was higher in the PEG-treated group compared with the control after 3 d of aerobic exposure. BA was detected only in the PEG-treated group upon aerobic exposure. Yeasts were more abundant in the control compared with the PEG-treated group after 7 d of aerobic exposure, after which the relative abundance of Lactobacillus was lower in the PEG-treated group (65.01% vs. 75.01% in the control; p < 0.05), while the relative abundance of Pediococcus was higher in the PEG-treated group compared with the control (10.9% vs. 4.49%, respectively; p < 0.05).The relative abundances of Apiotrichum and Aspergillus were lower in the control than in the PEG-treated group after 7 d of aerobic exposure. CONCLUSIONS: The results suggested that sainfoin CTs decreased aerobic stability, but could inhibit certain bacteria and fungi, such as Pediococcus and Apiotrichum, and preserve the protein content during the aerobic exposure of silage.

Effects of Bacillus subtilis or Lentilactobacillus buchneri on aerobic stability, and the microbial community in aerobic exposure of whole plant corn silage.

This study aimed to evaluate the effects of Bacillus subtilis or Lentilactobacillus buchneri on the fermentation quality, aerobic stability, and bacterial and fungal communities of whole plant corn silage during aerobic exposure. Whole plant corn was harvested at the wax maturity stage, which chopped to a length of approximately 1 cm, and treated with the following: distilled sterile water control, 2.0 × 105 CFU/g of Lentilactobacillus buchneri (LB) or 2.0 × 105 CFU/g of Bacillus subtilis (BS) for 42 days silage. Then, the samples were exposed to air (23-28°C) after opening and sampled at 0, 18 and 60 h, to investigate fermentation quality, bacterial and fungal communities, and aerobic stability. Inoculation with LB or BS increased the pH value, acetic acid, and ammonia nitrogen content of silage (P < 0.05), but it was still far below the threshold of inferior silage, the yield of ethanol was reduced (P < 0.05), and satisfactory fermentation quality was achieved. With the extension of the aerobic exposure time, inoculation with LB or BS prolonged the aerobic stabilization time of silage, attenuated the trend of pH increase during aerobic exposure, and increased the residues of lactic acid and acetic acid. The bacterial and fungal alpha diversity indices gradually declined, and the relative abundance of Basidiomycota and Kazachstania gradually increased. The relative abundance of Weissella and unclassified_f_Enterobacteria was higher and the relative abundance of Kazachstania was lower after inoculation with BS compared to the CK group. According to the correlation analysis, Bacillus and Kazachstania are bacteria and fungi that are more closely related to aerobic spoilage and inoculation with LB or BS could inhibit spoilage. The FUNGuild predictive analysis indicated that the higher relative abundance of fungal parasite-undefined saprotroph in the LB or BS groups at AS2, may account for its good aerobic stability. In conclusion, silage inoculated with LB or BS had better fermentation quality and improved aerobic stability by effectively inhibiting the microorganisms that induce aerobic spoilage.

Characterization of mycotoxins and microbial community in whole-plant corn ensiled in different silo types during aerobic exposure.

Silage can be contaminated with mycotoxins and accidental fungi after aerobic exposure. The study assessed the effects of bunker silos (BS), round bales (RB), and silage bags (SB) on the nutritional characteristics, fermentation quality, aerobic stability, mycotoxin levels and microbial communities of whole-plant corn silage (WPCS). After 90 days of fermentation, silages were opened and sampled at 0, 1, 3, 5, 7, and 9 days of exposure. SB group conserved higher lactic acid and dry matter contents and a lower pH value than other groups after 9 days of exposure (p < 0.05). The SB group showed the longest aerobic stability (202 h) among all silages (p < 0.05). The concentrations of aflatoxin B1, trichothecenes and fumonisin B1 were significantly lower in SB after 9 days of exposure (p < 0.05). Acetobacter became the dominant bacteria in BS and RB groups after 5 days of exposure. However, Lactobacillus still dominated the bacterial community in SB group. Acetobacter was positively correlated with pH, acetic acid content, and ammonia-N content (p < 0.05). Lactobacillus was positively correlated with Kazachstania and Candida abundances (p < 0.01) but negatively correlated with Fusarium abundance (p < 0.05). Considering the feed value and food safety of silage in the feeding process, silage bags are recommended for WPCS according to the observed nutritional quality, fermentation index and mycotoxin content.

Exploring the effect of wilting on fermentation profiles and microbial community structure during ensiling and air exposure of king grass silage.

In order to better understand the effect of wilting treatment on silage, we study analyzed the fermentation quality of unwilted (CK) and wilted (WT) king grass silage, and the dynamic changes of microorganisms in silage and aerobic exposure. After 30 days of silage, WT silage significantly reduced the pH of the silage (p < 0.05) and increased the contents of lactic acid and acetic acid (p < 0.05), but did not reduce the content of Ammonia-N (p > 0.05). Wilting treatment increased bacterial and fungal diversity during silage but decreased fungal diversity during aerobic exposure. The relative abundance of Lactococcus and Enterococcus in wilting silage increased. In the aerobic exposure stage, the relative abundance of Klebsiella decreased, but the relative abundance of Enterobacter increased in wilting treatment silage. In addition, the relative abundance of Acinetobacter and Ignatzschineria increased after 5 days of aerobic exposure. In contrast with unwilted silage, wilting treatment silage after aerobic exposure had no Candida, but the relative abundance of Wickerhamomyces increased. The results showed that wilting treatment could raise the silage quality of king grass. However, WT silage did not inhibit the reproduction of harmful microorganisms during aerobic exposure and did not significantly improve the aerobic stability of silage.

Dry matter recovery, ensiling characteristics and aerobic stability of oat silage treated with microbial inoculants at different temperatures.

To evaluate the effects of temperature and lactic acid bacteria (LAB) inoculants on oat silage in Loess Plateau of China, oat was harvested at dough stage, inoculated without (Control) or with LAB inoculants Synlac I (SLI, Lactobacillus plantarum and Pedioccocus acidilactici) and a selected strain HT1 (L. rhamnosus) and ensiled at 25°C (T25), 35°C (T35) and 45°C (T45). The fermentation quality was measured after 60 d of ensiling and the aerobic exposure was conducted at 30°C for 9 d. The results showed that control silage (stored at 25°C) had better fermentation quality than that ensiled at 35°C or 45°C. High temperature of 45°C resulted in sharp decreases in LAB counts and lactic acid concentration and increases in pH and NH3-N concentration in the control group. Inoculation improved the fermentation quality, and HT1 was more effective than SLI at 35°C and 45°C, while SLI showed better performance at 25°C. All silages displayed mild fluctuation for all treatments at the first 3 d of aerobic exposure, and significant differences were observed among treatments after that. Both control and inoculated silages stored at 25°C showed a sharp pH increase, while HT1 treated silages stored at 35°C and 45°C maintained stable pH and better fermentation quality during the aerobic exposure. In conclusion, SLI was suitable for oat silage fermentation at normal atmospheric temperature (25°C), while HT1 was more effective in improving DM recovery, fermentation quality and aerobic stability of oat silage at high temperature during summer in the Loess Plateau of China.

Effects of antibacterial peptide-producing Bacillus subtilis and Lactobacillus buchneri on fermentation, aerobic stability, and microbial community of alfalfa silage.

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.

Bacterial and fungal core microbiomes associated with small grain silages during ensiling and aerobic spoilage.

BACKGROUND: Describing the microbial populations present in small grain silage and understanding their changes during ensiling is of interest for improving the nutrient value of these important forage crops. Barley, oat and triticale forages as well as an intercropped mixture of the 3 crops were harvested and ensiled in mini silos for a period of 90 days, followed by 14 days of aerobic exposure. Changes in fermentation characteristics and nutritive value were assessed in terminal silages and bacterial and fungal communities during ensiling and aerobic exposure were described using 16S and 18S rDNA sequencing, respectively. RESULTS: All small grain silages exhibited chemical traits that were associated with well ensiled forages, such as low pH value (4.09 ± 0.28) and high levels of lactic acid (59.8 ± 14.59 mg/g DM). The number of microbial core genome operational taxonomic units (OTUs) decreased with time of ensiling. Taxonomic bacterial community profiles were dominated by the Lactobacillales after fermentation, with a notable increase in Bacillales as a result of aerobic exposure. Diversity of the fungal core microbiome was shown to also be reduced during ensiling. Operational taxonomic units assigned to filamentous fungi were found in the core microbiome at ensiling and after aerobic exposure, whereas the Saccharomycetales were the dominate yeast population after 90 days of ensiling and aerobic exposure. Bacterial and fungal orders typically associated with silage spoilage were identified in the core microbiome after aerobic exposure. CONCLUSION: Next Generation Sequencing was successfully used to describe bacterial communities and the first record of fungal communities throughout the process of ensiling and utilization. Adequately describing the microbial ecology of silages could lead to improved ensiling practices and the selection of silage inoculants that act synergistically with the natural forage microbiome.

Occurrence and stability of masked fumonisins in corn silage samples.

Corn plants contaminated with Fusarium verticilloides were harvested at two dates and ensiled in laboratory silos. The stability of Fumonisins B1 and B2 (FB1, FB2) was studied and the effects of inoculant use (Lactobacillus buchneri) and of the ensiling time were assessed. The occurrence of masked fumonisins was also investigated. After the extraction of the free forms (FB1, FB2), the residue was subjected to an alkaline hydrolysis resulting in the release of derivates subsequently detected by LC-HESI-MS/MS. The ensiling time has shown to be a significant factor for the evolution of free FB1 and FB2 at the 2nd harvest date. Moreover, the use of inoculant had a significant effect on the stability of hidden and total FB1 at the two harvest dates. Samples previously ensiled and exposed to aerobic conditions for 7days have shown higher fumonisin levels than those not exposed to air. Our results showed that 65% and 39% of the total FB1 and FB2 contents in silage were due to hidden fumonisins.