Terminal Silage Research Articles

Effects of a mixture of Lentilactobacillus hilgardii, Lentilactobacillus buchneri, Pediococcus pentosaceus and fibrolytic enzymes on silage fermentation, aerobic stability, and performance of growing beef cattle.

This study evaluated the effects of novel silage inoculants containing lactic acid bacteria (LAB) and fibrolytic enzymes on ensiling, aerobic stability (AS), and the performance of growing beef cattle. Whole-plant corn forage was either uninoculated (CON) or inoculated with a mixture of LAB containing (cfu g-1 fresh forage) 1.5 × 105 L. hilgardii (CNCM I-4785), 1.5 × 105 L. buchneri (NCIMB 40788) and 1.0 × 105 P. pentosaceus (NCIMB 12455) for a total of 4.0 × 105 cfu g-1 fresh forage LAB (IB), or a combination of IB plus fibrolytic enzymes (xylanase + β-glucanase) (IC). All treatments were ensiled in mini-silos, whereas CON and IC were also ensiled in silo bags for the growth performance study. Total bacteria (TB) counts were lower (P = 0.02) for IC than CON after 14 d of ensiling, whereas TB counts of IC and IB were greater (P ≤ 0.01) than CON after 60 d of ensiling in mini-silos. The LAB in IC and IB ensiled in mini-silos were greater than CON on d 60 (P ≤ 0.01) and 90 (P ≤ 0.001) of ensiling and after 3 d (P ≤ 0.01) of aerobic exposure (AE). Silage pH of IC ensiled in silo bags was lower than CON on d 3 (P < 0.01), 7 (P < 0.001), and 14 (P = 0.02) of AE. Yeast counts were lower for IC than CON in terminal silage (P < 0.001), and after 3 (P < 0.001) and 7 d (P < 0.01) of AE. Acetate (AC) concentrations were higher (P ≤ 0.02) for IC than CON throughout AE, whereas lactate (LA) concentrations of IC were greater than CON on d 3 (P < 0.001), 7 (P < 0.01), and 14 (P < 0.001) of AE. Greater AC concentration and lower yeast counts resulted in greater (P < 0.001) stability for IC ensiled in silo bags than CON after 14 d of AE. Growth performance of steers was similar (P > 0.05) as the nutrient composition of silage was similar across diets. Improved AS of IC could potentially have a greater impact on DMI, production efficiency, and growth performance in large-scale commercial feedlot operations where silage at the silo face may be exposed to air for longer periods of time.

Exploring the Epiphytic Microbial Community Structure of Forage Crops: Their Adaptation and Contribution to the Fermentation Quality of Forage Sorghum during Ensiling.

In this study, the effects of epiphytic microbiota from different forages on the fermentation characteristics and microbial community structure of forage sorghum silage were investigated. The gamma irradiated sterilized forage sorghum was treated through sterile water, epiphytic microbiota of forage sorghum (FSm), Sudan grass (SDm), Napier grass (NPm), and maize (MZm). NPm and SDm inoculated silages showed similar pH value and lactic acid (LA) and acetic acid (AA) contents at day 3 and 60 of ensiling. The final silage of FSm and MZm showed lower (p < 0.05) pH and AA content and a higher LA content compared to the NPm and SDm silages. Bacterial species from the Weisella genus were predominantly present in FSm, NPm, and SDm, while Lactococcus dominated the MZm silage during early ensiling. Lactobacillus was predominant in all inoculated terminal silages. Overall, the four inoculated microbiota decreased the pH value of silage and were dominated by lactic acid bacteria (LAB); however, the NPm and SDm treatments resulted in comparatively higher AA contents which could have an inhibitory effect on the secondary fermentation developed by the yeast and enhanced the aerobic stability of forage sorghum silage.

Using a high-throughput sequencing technology to evaluate the various forage source epiphytic microbiota and their effect on fermentation quality and microbial diversity of Napier grass.

Napier grass (Pennisetum purpureum) is well-known due to its high biomass production. The epiphytic microbiota was prepared from Napier, alfalfa, and red clover grass and served as an inoculum. The chopped sterilized Napier grass was inoculated with reconstituted epiphytic microbiota, and treatments were designed as: distilled water (N0); Napier grass epiphytic microbiota (NP); alfalfa epiphytic microbiota (AL); and red clover epiphytic microbiota (RC). The results reveal that the reconstituted epiphytic microbiota bacteria efficiently adapted in Napier grass silage, improved fermentation, and produced lactic acid. The alfalfa-grass inoculum rapidly dropped pH and enhanced the lactic acid (LA) and the ratio of lactic acid-to-acetic acid (LA/AA) during the entire ensiling process. However, red clover attains high lactic acid, while Napier grass produces high acetic acid-type fermentation at terminal silage. After day 60 of ensiling, Lactobacillus proportion was higher in AL (85.45%), and RC (59.44%), inocula as compared with NP (36.41%), inoculum. The NP inoculum terminal silage was diverse than AL and RC inocula and dominated by Enterobacter (16.32%) and Enterobacteriaceae (10.16%) and also significantly (p < 0.05) higher in acetic acid. The present study concluded that AL and RC epiphytic microbiota successfully develop and more efficient than Napier grass microbiota. It is suggested that abundant microbiota isolate from alfala and red clover and develop more economical and efficient inocula for quality fermentation of Napier grass silage in practice.

Abundance and diversity of epiphytic microbiota on forage crops and their fermentation characteristic during the ensiling of sterile sudan grass.

This study aimed to evaluate the effects of exogenous epiphytic microbiota inoculation on the fermentation quality and microbial community of sudan grass silage. Gamma irradiated sudan grass was ensiled with distilled water (STR), epiphytic microbiota of sudan grass (SUDm), forage sorghum (FSm), napier grass (NAPm) and whole crop corn (WCCm). The FSm inoculated silage have significantly lower lactic acid (LA) concentration and higher pH during early ensiling, while LA concentration gradually and significantly increased with the progression of ensiling and have lower pH in relation to other treatments for terminal silage. Inoculation of NAPm resulted in lower LA and higher acetic acid (AA) concentrations, higher pH, ammonia-N and dry matter losses for terminal silage, followed by SUDm silage. Inoculations of WCCm significantly increased LA production and pH decline during early ensiling and have higher LA and pH then NAPm and SUDm silages during final ensiling. The early fermentation of SUDm silage was dominated by genus of Pediococcus. The genera of Lactobacillus were predominant in WCCm and NAPm silages during 3days of ensiling, while Weissella dominated initial microbial community of FS silage. The terminal silage of NAPm was dominated by Enterobacter and Rosenbergiella, while Enterobacter and Lactobacillus dominated terminal SUDm silage. The final silage of FSm was dominated by Lactobacillus, Weissella and Pediococcus, while Lactobacillus and Acetobacter dominated terminal WCCm silages. The results demonstrated that among the four forages the epiphytic microbiota from forage sorghum positively influenced the microbial community and fermentability of sudan grass silage.

Exploring microbial community structure and metabolic gene clusters during silage fermentation of paper mulberry, a high-protein woody plant

To develop a new high-protein woody forage resource for livestock to alleviate feed shortages in the tropics, we applied PacBio single-molecule, real-time (SMRT) sequencing to explore the community structure, species diversity and metabolic gene clusters of natural microorganisms associated with paper mulberry (PM) silage fermentation. High levels of microbial diversity and abundance were observed in PM raw material, and these levels decreased with the progression of silage fermentation. During woody ensiling, the dominant bacteria shifted from pathogenic Gram-negative Proteobacteria to beneficial Gram-positive Firmicutes. Lactic acid bacteria became the most dominant bacteria that affected fermentation quality in terminal silages. Global and overview maps, carbohydrate metabolism and amino-acid metabolism were the important microbial metabolic pathways that impacted the final fermentation product of silage. PacBio SMRT sequencing revealed specific microbial-related information concerning silage. PM is rich in nutrients and macro mineral contents, which are preserved well during ensiling, indicating that PM silage can serve as a new woody resource suitable for ruminants.

Effects of inoculation of corn silage with Lactobacillus hilgardii and Lactobacillus buchneri on silage quality, aerobic stability, nutrient digestibility, and growth performance of growing beef cattle.

This study evaluated the effects of inoculation of whole crop corn silage with a mixture of heterofermentative lactic acid bacteria (LAB) composed of Lactobacillus hilgardii and Lactobacillus buchneri on ensiling, aerobic stability, ruminal fermentation, total tract nutrient digestibility, and growth performance of beef cattle. Uninoculated control corn silage (CON) and silage inoculated with 3.0 × 105 cfu g-1 of LAB containing 1.5 × 105 cfu g-1 of L. hilgardii CNCM I-4785 and 1.5 × 105 cfu g-1 of L. buchneri NCIMB 40788 (INOC) were ensiled in silo bags. The pH did not differ (P > 0.05) between the two silages during ensiling but was greater (P < 0.001) for CON than INOC after 14 d of aerobic exposure (AE). Neutral detergent insoluble crude protein (NDICP) content (% of DM and % of CP basis) of terminal INOC silage was greater (P ≤ 0.05) than that of CON. In terminal silage, concentrations of total VFA and acetate were greater (P < 0.001), while water-soluble carbohydrates were lower (P < 0.001) for INOC than CON. Yeast and mold counts were lower for INOC than CON (P ≤ 0.001) in both terminal and aerobically exposed silages. The stability of INOC was greater (P < 0.001) than that of CON after 14 d of AE. Ruminal fermentation parameters and DMI did not differ (P > 0.05) between heifers fed the two silages, while there was a tendency (P ≤ 0.07) for lower CP and starch digestibility for heifers fed INOC than CON. Total nitrogen (N) intake and N retention were lower (P ≤ 0.04) for heifers fed INOC than CON. Dry matter intake as a percentage of BW was lower (P < 0.04) and there was a tendency for improved feed efficieny (G:F; P = 0.07) in steers fed INOC vs. CON silage. The NEm and NEg contents were greater for INOC than CON diets. Results indicate that inoculation with a mixture of L. hilgardii and L. buchneri improved the aerobic stability of corn silage. Improvements in G:F of growing steers fed INOC silage even though the total tract digestibility of CP and starch tended to be lower for heifers fed INOC are likely because the difference in BW and growth requirements of these animals impacted the growth performance and nutrient utilization and a greater proportion of NDICP in INOC than CON.

Microbial diversity and fermentation profile of red clover silage inoculated with reconstituted indigenous and exogenous epiphytic microbiota

The study investigated the effects of transplantation and reconstitution of indigenous and exogenous epiphytic microbiota on the fermentation quality and microbial community of red clover silage. Sterile red clover was inoculated with distilled water (RC0), extracted epiphytic microbiota of red clover (RC), maize (MZ), and sorghum (SG). RC inoculation rapidly decreased pH at the onset of ensiling. The LA concentration and ratio of LA/AA were higher in RC silage during entire ensiling while MZ silage during late phase of ensiling. Pediococcus was dominant in RC early silage, while Lactobacillus was abundant in MZ final silage. The SG terminal silage had higher pH (>4.50) and dominated by Sphingomonas, Enterobacter, and Novosphingobium. RC and MZ microbiota were beneficial in enhancing fermentation quality and microbial community in red clover silage. Transplantation and reconstitution of epiphytic microbiota can be a successful method to assess the effective and eco-friendly additive for the targeted crop.

The feasibility and effects of exogenous epiphytic microbiota on the fermentation quality and microbial community dynamics of whole crop corn

This study analyzed the feasibility and effects of exogenous epiphytic microbiota on the fermentation quality and microbial community of whole crop corn. Gamma irradiated whole crop corn was ensiled with distilled water (STR), extracted microbiota of whole crop corn (WCC), Napier grass (NAP), forage sorghum (FS) or Sudan grass (SUD). WCC significantly increased LA concentration and decreased the pH during early ensiling. FS had significantly higher LA and lower pH during terminal ensiling. NAP caused higher pH and AA followed by the SUD silage. During 3 d of ensiling WCC and FS silage was dominated by Lactobacillus and Lactococcus while Weissella dominated NAP and SUD silage. Terminal silage was dominated by Lactobacillus in FS and Acinetobacter in NAP while Lactobacillus, Acetobacter, Acinetobacter and Sphingobium dominated WCC and SUD silage. The study demonstrated that FS microbiota transplantation positively influenced the microbial community and fermentation quality of whole crop corn silage as compared to other microbiota.

Effects of inoculation of corn silage with Lactobacillus spp. or Saccharomyces cerevisiae alone or in combination on silage fermentation characteristics, nutrient digestibility, and growth performance of growing beef cattle.

This study evaluated the effects of a novel silage inoculant containing Saccharomyces cerevisiae strain 3 as a direct fed microbial (DFM) on the ensiling, aerobic stability, and nutrient digestibility of whole-crop corn silage and growth performance of beef cattle. Treatments included uninoculated corn silage (CON) or corn silage inoculated with a mixture of 1.1 × 105 cfu g-1 fresh forage Lactobacillus plantarum and Lactobacillus buchneri (INOC1) or 1.0 × 104 cfu g-1 fresh forage S. cerevisiae strain 3 (INOC2) or a mixture of INOC1 and INOC2 (INOC3). Silage in INOC1 had lower (P = 0.03) proportion of lactate, with acetate (Ac) proportion ranking as INOC1 > INOC3 > INOC2 (P < 0.01). In terminal silage, numbers of lactic acid bacteria were greater (P = 0.05) for INOC1 than CON and INOC2, while yeast counts tended (P = 0.08) to be greater for INOC2 than INOC3 on day 3 of aerobic exposure. Aerobic stability of corn silage was not impacted by inoculation with S. cerevisiae strain 3. Heifers fed INOC2 and INOC3 had lower (P < 0.01) ruminal Ac concentration than those fed CON. Apparent total tract digestibilities of DM, OM, ADF, and NDF were greater (P ≤ 0.03) for heifers fed INOC2 than those fed CON. Growth performance was similar across treatments, excepting DMI as percent of BW tended to be lower (P = 0.08) for INOC2 steers compared to CON steers. These results suggest that S. cerevisiae strain 3 has potential as a component in a fourth generation DFM silage inoculant.

Impact of Saccharomyces cerevisiae and Lactobacillus buchneri on microbial communities during ensiling and aerobic spoilage of corn silage1.

The objective of this study was to assess the impact of Saccharomyces cerevisiae in combination with Lactobacillus buchneri on the fermentation characteristics, aerobic stability, nutritive value, and microbial communities of corn silage. Whole crop corn (39% DM) was either uninoculated (Control) or inoculated with S. cerevisiae and L. buchneri at the following concentrations: S. cerevisiae 104 cfu/g fresh forage (S4), S. cerevisiae 105 cfu/g (S5), S. cerevisiae 104 cfu/g + L. buchneri 105 cfu/g (S4L5), and S. cerevisiae 105 cfu/g + L. buchneri 104 cfu/g (S5L4), and ensiled in mini silos for 118 d, followed by 7 d of aerobic exposure. Changes in fermentation characteristics and nutritive value were assessed in terminal silages. Saccharomyces cerevisiae, L. buchneri, and total yeast, fungal, and bacterial communities in silage were estimated using quantitative PCR. Composition of bacterial and fungal communities during ensiling and aerobic exposure was measured using 16S rDNA and ITS sequencing, respectively. In the first 7 d of ensiling, the concentration of lactic acid rapidly increased (P < 0.01) in all silages, with the pH declining to 4.0 (P < 0.001) and thereafter remaining stable (P = 0.23). Although S4L5 contained a higher (P = 0.03) concentration of acetic acid than Control, other fermentation characteristics were did not differ among terminal silages. Inoculation with S. cerevisiae had no detrimental effect on the aerobic stability of silage, whereas L. buchneri did not prevent spoilage as the pH across all silages averaged 8.0 after 7 d of aerobic exposure. Total yeast (P = 0.42), bacterial (P = 0.13), and fungal (P = 0.89) communities were not altered by the inoculants after ensiling or aerobic exposure. Sequencing identified temporal shifts of bacterial and fungal communities during ensiling and aerobic exposure. Concentrations of S. cerevisiae and L. buchneri in all inoculated silages remained greater (P < 0.01) than Control after ensiling, with numbers of S. cerevisiae increasing after 7 d of aerobic exposure. Bacterial communities in silages inoculated with S. cerevisiae and L. buchneri clustered separately from other silages, an observation that was not apparent for fungal communities. Our findings suggest that aerobic exposure could potentially increase the abundance of S. cerevisiae with probiotic properties in corn silage just prior to feeding.

Condensed Tannins Affect Bacterial and Fungal Microbiomes and Mycotoxin Production during Ensiling and upon Aerobic Exposure.

Purple prairie clover (PPC; Dalea purpurea Vent.) containing 84.5 g/kg dry matter (DM) of condensed tannin (CT) was ensiled without (control) or with polyethylene glycol (PEG) for 76 days, followed by 14 days of aerobic exposure. Changes in fermentation characteristics were determined, and the composition of bacterial and fungal communities were assessed using metagenomic sequencing. The addition of polyethylene glycol (PEG) that deactivated CT at ensiling increased (P < 0.05 to ∼0.001) soluble N, nonprotein N, lactic acid, total volatile fatty acids, ammonia N, deoxynivalenol (DON), and ochratoxin A (OTA) but decreased (P < 0.001) pH and water-soluble carbohydrates. The concentrations of DON and OTA increased (P < 0.001) for both silages, with the extent of increase being greater for control than for PEG-treated silage during aerobic exposure. The PEG-treated silage exhibited higher (P < 0.01 to ∼0.001) copy numbers of total bacteria, Lactobacillus, yeasts, and fungi than the control. The addition of PEG decreased (P < 0.01) bacterial diversity during both ensiling and aerobic exposure, whereas it increased (P < 0.05) fungal diversity during aerobic exposure. The addition of PEG at ensiling increased (P < 0.05) the abundances of Lactobacillus and Pediococcus species but decreased (P < 0.01) the abundances of Lactococcus and Leuconostoc species. Filamentous fungi were found in the microbiome at ensiling and after aerobic exposure, whereas Bacillus spp. were the dominate bacteria after aerobic exposure. In conclusion, CT decreased protein degradation and improved the aerobic stability of silage. These desirable outcomes likely reflect the ability of PPC CT to inhibit those microorganisms involved in lowering silage quality and in the production of mycotoxins.IMPORTANCE The present study reports the effects of condensed tannins on the complex microbial communities involved in ensiling and aerobic exposure of purple prairie clover. This study documents the ability of condensed tannins to lower mycotoxin production and the associated microbiome. Taxonomic bacterial community profiles were dominated by Lactobacillales after fermentation, with a notable increase in Bacillus spp. as a result of aerobic exposure. It is interesting to observe that condensed tannins decreased bacterial diversity during both ensiling and aerobic exposure but increased fungal diversity during aerobic exposure only. The present study indicates that the effects of condensed tannins on microbial communities lead to reduced lactic acid and total volatile fatty acid production, proteolysis, and mycotoxin concentration in the terminal silage and improved aerobic stability. Condensed tannins could be used as an additive to control unfavorable microbial development and maybe enhanced feed safety.

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

BackgroundDescribing 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.ResultsAll 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.ConclusionNext 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.

THE GENERAL PATTERN IN SILAGE FERMENTATION IN TWO SUBTROPICAL GRASSES

Three sets of previously‐published experiments have been re‐analysed by numerical methods intended for the elucidation of pattern in sequential data. The results suggest that for Setaria sphacelata cv. Nandi and Chloris gayana cv. Samford Rhodes the stable terminal silage to be expected under subtropical conditions is not the high lactic‐acid silage of temperate regions, but a silage with a relatively high proportion of acetic acid. Lactic‐acid silage is formed, but usually behaves as though in a transitional phase. However, silage made from mature Setaria can be maintained in the lactic‐acid phase by the addition of high percentages of sugar. The silage fermentation can sometimes be retarded, or deflected into an undesirable path for reasons that are not understood.The above patterns suggest three hypotheses for more fundamental investigation. First, since acetic‐acid silage can be expected under most subtropical conditions, its properties require careful investigation. Secondly, relatively young Setaria may be regarded as a source of acetic‐acid silage, but the optimum amount of sugar to be added for ensiling more mature grass needs investigation. Thirdly there is an urgent need to investigate the causes of retardation of fermentation, and of its deflection into an undesirable path, because until these causes are understood, the outcome of ensilage will remain uncertain.