The effects of Lactobacillus hilgardii 4785 and Lactobacillus buchneri 40788 on the microbiome,fermentation, and aerobic stability of corn silage ensiled for various times

We evaluated the ability of an inoculant containing a combination of Lactobacillus hilgardii and Lactobacillus buchneri to modify the microbiome and improve the aerobic stability of whole-plant corn silage after various lengths of ensiling. Chopped whole-plant corn at about 33% dry matter (DM) was uninoculated (CTR) or inoculated with L. hilgardii CNCM I-4785 and L. buchneri NCIMB 40788 at 200,000 cfu/g of fresh forage weight each (combined application rate of 400,000 cfu of lactic acid bacteria/g of fresh forage weight; LHLB), L. buchneri NCIMB 40788 at 400,000 cfu/g of fresh forage weight and Pediococcus pentosaceus NCIMB 12455 at 100,000 cfu/g of fresh forage weight, used as a positive control (LB500), L. hilgardii CNCM I-4785 at the application rate used in the LHLB formulation of 200,000 cfu/g of fresh forage weight (LH), or L. buchneri NCIMB 40788 at the application rate used in the LHLB formulation of 200,000 cfu/g of fresh forage weight (LB). Silos were opened after 34 and 99 d of ensiling and analyzed for nutrient composition, fermentation profile, microbiome, and aerobic stability. After 34 d of ensiling, the inoculated silages had greater numbers of culturable lactic acid bacteria, a bacterial community less rich and diverse, greater relative abundance of Lactobacillus, lower relative abundance of Klebsiella, and a greater concentration of propionic acid than uninoculated silages. Inoculation decreased the ratio of lactic acid to acetic acid, except for LB alone. Treatment LHLB resulted in silage with a greater concentration of 1,2-propanediol than LB500 and was the only treatment to have a lower relative abundance of Saccharomycetes compared with uninoculated silage. Treatments LHLB and LB500 improved the aerobic stability compared with CTR, but the individual LH and LB treatments applied at a low dose did not. Whereas LB500 was stable 34 h longer than CTR, LHLB was stable 91 h longer. After 99 d of ensiling, all inoculated silages had markedly greater aerobic stability than uninoculated silage and were stable for more than 360 h. The inoculant containing a combination of L. hilgardii and L. buchneri markedly improved the aerobic stability of corn silage after a relatively short period of ensiling, and such improvements were greater than the ones obtained from inoculation with the combination of L. buchneri and P. pentosaceus. Inoculating with the combination of L. hilgardii and L. buchneri may be helpful to producers that must feed silage shortly after ensiling.     

Dry matter and nutritional losses during aerobic deterioration of corn and sorghum silages as influenced by different lactic acid bacteria inocula

The economic damage that results from aerobic deterioration of silage is a significant problem for farm profitability and feed quality. This paper quantifies the dry matter (DM) and nutritional losses that occur during the exposure of corn and sorghum silages to air over 14 d and assesses the possibility of enhancing the aerobic stability of silages through inoculation with lactic acid bacteria (LAB). The trial was carried out in Northern Italy on corn (50% milk line) and grain sorghum (early dough stage) silages. The crops were ensiled in 30-L jars, without a LAB inoculant (C), with a Lactobacillus plantarum inoculum (LP), and with a Lactobacillus buchneri inoculum (LB; theoretical rate of 1 × 10⁶ cfu/g of fresh forage). The pre-ensiled material, the silage at silo opening, and the aerobically exposed silage were analyzed for DM content, fermentative profiles, yeast and mold count, starch, crude protein, ash, fiber components, 24-h and 48-h DM digestibility and neutral detergent fiber (NDF) degradability. The yield and nutrient analysis data of the corn and sorghum silages were used as input for Milk2006 to estimate the total digestible nutrients, net energy of lactation, and milk production per Mg of DM. The DM fermentation and respiration losses were also calculated. The inocula influenced the in vitro NDF digestibility at 24 h, the net energy for lactation (NE_L), and the predicted milk yield per megagram of DM, whereas the length of time of air exposure influenced DM digestibility at 24 and 48 h, the NE_L, and the predicted milk yield per megagram of DM in the corn silages. The inocula only influenced the milk yield per megagram of DM and the air exposure affected the DM digestibility at 24 h, the NE_L, and the milk yield per megagram of DM in the sorghum silages. The milk yield, after 14 d of air exposure, decreased to 1,442, 1,418, and 1,277 kg/Mg of DM for C, LB, and LP corn silages, respectively, compared with an average value of 1,568 kg of silage at opening. In the sorghum silages, the milk yield, after 14 d of air exposure, decreased to 1,226, 1,278, and 1,250 kg/Mg of DM for C, LB, and LP, respectively. When the estimated milk yield per megagram of harvested DM of corn and sorghum silage were related to mold count, it was shown that the loss of potential milk production occurred when the mold count exceeded 4 log cfu/g of silage, and it was almost halved when the mold count reached values greater than 8 log cfu/g of silage. Inoculation with L. buchneri, at a rate of 1 × 10⁶ cfu/g of fresh forage, enhanced the stability of the silage after exposure to air, and, consequently, contributed to maintaining the nutritional value of the harvested forage over time, for air exposure up to 7 d.     

Increase in aflatoxins due to Aspergillus section Flavi multiplication during the aerobic deterioration of corn silage treated with different bacteria inocula

The growth of Aspergillus flavus and the production of aflatoxins (AF) during the aerobic deterioration of corn silage represent a problem for animal and human health. This experiment was conducted to evaluate whether the presence of A. flavus and AF production originate from the field or additional AF are produced during the fermentation phase or during aerobic deterioration of corn silage. The trial was carried out in northern Italy on corn at a dry matter (DM) level of 34%. The fresh herbage was either not treated (C) or treated with a Lactobacillus buchneri (LB) NCIMB 40788 [(at 3 × 10⁵ cfu/g of fresh matter (FM)], Lactobacillus hilgardii (LH) CNCM I-4785 (at 3 × 10⁵ cfu/g of FM), or their combination (LB+LH; at 1.5 × 10⁵ cfu/g of FM of each strain) ensiled in 20-L silos and opened after 250 d of ensiling. After silo opening, the aerobic stability was evaluated and samples were taken after 7 and 14 d of air exposure. The pre-ensiled material, the silages at silo opening, and the aerobically exposed silages were analyzed for DM content, fermentative profiles, microbial count, nutritive characteristics, DM losses, and AFB₁, AFB₂, AFG₁, and AFG₂ contents. Furthermore, a subsample of colonies with macromorphological features of Aspergillus section Flavi was selected for AF gene pattern characterization and in vitro AF production. The presence of A. flavus was below the detection limit (<1.00 log₁₀ cfu/g) in the fresh forage before ensiling, whereas it was found in 1 out of 16 silage samples at silo opening at a level of 1.24 log₁₀ cfu/g. The AF were found in both the fresh forage and at opening in all the samples, with a predominance of AFB₂ (mean value of 1.71 μg/kg of DM). The inoculation of lactic acid bacteria determined a reduction in the lactic-to-acetic ratio compared with the control. A larger amount of acetic acid resulted in a lower yeast count and higher aerobic stability in the treated silages than in the control ones. At the beginning of aerobic deterioration, the yeasts increased to over 5 log₁₀ cfu/g, whereas the molds were close to the value observed at silo opening. When the inhibiting conditions were depleted (pH and temperature higher than 5 and 35°C, respectively), both the total molds and A. flavus reached higher values than 8.00 and 4.00 log₁₀ cfu/g, respectively, thus determining the ex novo production of AFB₁ during aerobic deterioration, regardless of treatments. The analysis of gene pattern showed that 64% of the selected colonies of A. flavus showed the presence of all 4 AF gene patterns, and 43% of the selected colonies were able to produce AF in vitro. During air exposure, after 1,000°C·h have been cumulated, starch content decreased (below 10% DM) and concentration of neutral detergent fiber, acid detergent fiber, hemicelluloses, crude protein, and ash increased. The inoculation with LB and LB+LH increased the aerobic stability of the silages and delayed the onset of aerobic microbial degradation, which in turn indirectly reduced the risk of A. flavus outgrowth and AFB₁ production after silage opening.     

Conservation characteristics of corn ears and stover ensiled with the addition of Lactobacillus plantarum MTD-1, Lactobacillus plantarum 30114, or Lactobacillus buchneri 11A44

The aim of this study was to investigate the effects of inoculating 3 contrasting lactic acid bacteria on the fermentation profile, estimated nutritive value, and aerobic stability of corn ears and stover produced under marginal growing conditions. Ears and stover were separated from whole-crop corn plants obtained from 3 replicate field blocks. Representative subsamples were precision chopped and allocated to 1 of the following treatments: an uninoculated control, Lactobacillus plantarum MTD-1 (LP1), L. plantarum 30114 (LP2), or Lactobacillus buchneri 11A44 (LB). Each bacterial additive was applied at a rate of 1 × 10(6) cfu/g of fresh herbage. Triplicate samples of each treatment were ensiled in laboratory silos at 15°C for 3, 10, 35, or 130 d. No difference was observed between the dry matter recoveries of uninoculated ear or stover silages and silages made with LP1, and the aerobic stability of uninoculated ear and stover silages did not differ from silages made with LB. Stover silages made with LP2 and ensiled for 35 d had a lower proportion of lactic acid in total fermentation products compared with LP1. The aerobic stability and dry matter recovery of ear and stover silages in this study were not improved when made with LB, LP1, or LP2, due to the indigenous highly heterolactic fermentation that prevailed in the uninoculated ear and stover during 130-d ensilage.     

Fermentation and aerobic stability of rehydrated corn grain silage treated with different doses of Lactobacillus buchneri or a combination of Lactobacillus plantarum and Pediococcus acidilactici

We investigated the effects of different types and doses of inoculants for ensiling rehydrated corn grain. Shelled corn was finely ground and rehydrated to 35% moisture. Treatments were as follows: (1) control (no additives); (2) Lactobacillus plantarum and Pediococcus acidilactici (LPPA) at a theoretical application rate of 1 × 10⁵ cfu/g; (3) LPPA at 5 × 10⁵ cfu/g; (4) LPPA at 1 × 10⁶ cfu/g; (5) Lactobacillus buchneri (LB) at 1 × 10⁵ cfu/g; (6) LB at 5 × 10⁵ cfu/g; and (7) LB at 1 × 10⁶ cfu/g. We detected no effect of inoculant dose. Gas losses were greater in silages treated with LB compared with control and LPPA silages. Treating silages with LB reduced the concentrations of lactic acid and ethanol and increased silage pH and concentrations of acetic acid, propionic acid, and 1,2-propanediol. At silo opening, silages treated with LB had higher counts of lactic acid bacteria but lower yeast counts than the control silage. Aerobic stability was greater for silages treated with LB and lower for silages treated with LPPA compared with the control. The LB reduced dry matter (DM) losses during aerobic exposure, whereas LPPA increased them. Prolamin content was lower in silages treated with LB compared with the control, resulting in greater ruminal in situ DM degradability. Inoculating LB to a dose of 1 × 10⁵ cfu/g increased aerobic stability and ruminal in situ DM degradability of rehydrated corn grain silage. The addition of LPPA did not alter the fermentation process and worsened the aerobic stability of rehydrated corn grain silage. Further studies are warranted to confirm these conclusions in other corn hybrids, inoculants, and their combinations.     

Effect of Lactobacillus hilgardii,Lactobacillus buchneri,or their combination on the fermentation and nutritive value of sorghum silage and corn silage

The objective of this study was to determine the effect of inoculation with Lactobacillus hilgardii with or without Lactobacillus buchneri on the fermentation, chemical composition, and aerobic stability of sorghum and corn silage after 2 ensiling durations. Sorghum forage was harvested at 27% dry matter (DM; experiment 1), and different corn hybrids were harvested at late (43.8% DM; experiment 2) or normal maturity (34% DM; experiment 3). All harvested forages were chopped and ensiled in quadruplicate in vacuum-sealed nylon-polyethylene bags (40 × 61 cm) for 30 and 90 d after treatment with (1) deionized water (uninoculated) or (2) L. buchneri (1.5 × 10⁵ cfu/g of fresh weight; LB); (3) L. hilgardii (1.5 × 10⁵ cfu/g of fresh weight; LH); or (4) L. buchneri and L. hilgardii (1.5 × 10⁵ cfu/g of fresh weight of each inoculant). Data for each experiment were analyzed separately accounting for the 2 × 2 × 2 factorial treatment arrangement. Inoculating sorghum forage with LB or LH separately increased acetate and 1,2 propanediol concentration, tended to increase DM loss, reduced lactate concentration and the lactate-to-acetate ratio, and increased aerobic stability after 90 but not after 30 d of ensiling. Inoculating late-harvested corn silage with LB or LH separately increased and decreased DM loss, respectively, increased 1,2 propanediol concentration, reduced lactate-to-acetate ratio and yeast counts but did not affect aerobic stability. Inoculating normal-harvested corn silage with LH reduced DM loss and increased 1,2 propanediol concentration and yeast counts; LB reduced lactate concentration, lactate-to-acetate ratio, and total acids. Either inoculant alone increased aerobic stability after 30 or 90 d. The main benefit of combining LB with LH was prevention of increases in DM losses by LH or LB separately. No improvement in aerobic stability resulted from applying LH instead of LB separately or from combining them. Application of LB or LH separately improved aerobic stability of sorghum silage after 90 d and normal-harvested corn silage after 30 or 90 d but did not affect that of late-harvested corn silage.     

The effect of Lactobacillus buchneri, Lactobacillus plantarum, or a chemical preservative on the fermentation and aerobic stability of corn silage

Several microorganisms and one chemical preservative were tested for their effects on the fermentation and aerobic stability of corn silage. Whole-plant corn (one-half milk line, 31.3% dry matter) was ensiled in quadruplicate 20-L laboratory silos untreated or after the following treatments: Lactobacillus buchneri at 1 x 10(5) and 1 x 10(6) cfu/g of fresh forage; two different strains of L. plantarum, each at 1 x 10(6) cfu/g; and a buffered propionic acid-based product at 0.1% of fresh forage weight. After 100 d of ensiling, silage treated with L. buchneri (1 x 10(6) cfu/g) had a lower concentration of lactic acid compared with the untreated silage, but was similar to other treated silages. The silage treated with the high (1 x 10(6) cfu/g), but not the moderate rate (1 x 10(5) cfu/g) of L. buchneri also had a greater concentration of acetic acid (3.60%) and less yeasts (2.01 log cfu/g) when compared with other treatments (average of 1.88% acetic acid and 5.85 log cfu of yeasts/g). Silages treated with L. plantarums, the moderate rate of L. buchneri, and the chemical preservative took longer to heat than untreated silage when exposed to air, but improvements were numerically small (6.3 to 10.5 h). In contrast, silage treated with the high rate of L. buchneri never heated throughout a 900-h period of monitoring. Inoculating corn silage with 1 x 10(6) cfu/g of L. buchneri resulted in a more heterolactic fermentation and dramatically improved the aerobic stability of corn silage.     

The effect of treating whole-plant barley with Lactobacillus buchneri 40788 on silage fermentation,aerobic stability,and nutritive value for dairy cows

Chopped barley forage was ensiled untreated or treated with several doses (1 x 10(5) to 1 x 10(6) cfu/g of fresh forage) of Lactobacillus buchneri 40788 in laboratory silos and untreated or treated (4 x 10(5) cfu/g) in a farm silo. Silage from the farm silos was fed to lactating cows. In the laboratory silo, the effects of inoculation on fermentation and aerobic stability were also compared to silage treated with a commercial inoculant and a buffered propionic acid additive. Inoculation with L. buchneri 40788 decreased the final concentrations of lactic acid but increased concentrations of acetic acid and ethanol in silage from laboratory and farm silos. Silages stored in laboratory silos did not heat after exposure to air for 7 d and were then mixed with alfalfa silage and a concentrate to form total mixed rations (TMR) that were further exposed to air. The TMR containing silages treated with L. buchneri 40788 or a buffered propionic-acid-based additive took longer to heat and spoil than the TMR containing untreated silage or silagetreated with the commercial inoculant. Silage stored in a farm silo and treated with L. buchneri 40788 had fewer yeasts and molds than did untreated silage. Aerobic stability was greater in treated silage alone and in a TMR containing treated silage. Dry matter intake (18.6 kg/d), milk production (25.7 kg/d), and milk composition did not differ between cows fed a TMR containinguntreated or treated silage. These findings show that L. buchneri can improve the aerobic stability of barley silage in laboratory and farm silos and that feeding treated silage had no negative effect on intake or performance.     

Meta-analysis of effects of inoculation with homofermentative and facultative heterofermentative lactic acid bacteria on silage fermentation,aerobic stability,and the performance of dairy cows

Forages are usually inoculated with homofermentative and facultative heterofermentative lactic acid bacteria (LAB) to enhance lactic acid fermentation of forages, but effects of such inoculants on silage quality and the performance of dairy cows are unclear. Therefore, we conducted a meta-analysis to examine the effects of LAB inoculation on silage quality and preservation and the performance of dairy cows. A second objective was to examine the factors affecting the response to silage inoculation with LAB. The studies that met the selection criteria included 130 articles that examined the effects of LAB inoculation on silage quality and 31 articles that investigated dairy cow performance responses. The magnitude of the effect (effect size) was evaluated using raw mean differences (RMD) between inoculated and uninoculated treatments. Heterogeneity was explored by meta-regression and subgroup analysis using forage type, LAB species, LAB application rate, and silo scale (laboratory or farm-scale) as covariates for the silage quality response and forage type, LAB species, diet type [total mixed ration (TMR) or non-TMR], and the level of milk yield of the control cows as covariates for the performance responses. Inoculation with LAB (≥10⁵ cfu/g as fed) markedly increased silage fermentation and dry matter recovery in temperate and tropical grasses, alfalfa, and other legumes. However, inoculation did not improve the fermentation of corn, sorghum, or sugarcane silages. Inoculation with LAB reduced clostridia and mold growth, butyric acid production, and ammonia-nitrogen in all silages, but it had no effect on aerobic stability. Silage inoculation (≥10⁵ cfu/g as fed) increased milk yield and the response had low heterogeneity. However, inoculation had no effect on diet digestibility and feed efficiency. Inoculation with LAB improved the fermentation of grass and legume silages and the performance of dairy cows but did not affect the fermentation of corn, sorghum, and sugar cane silages or the aerobic stability of any silage. Further research is needed to elucidate how silage inoculated with homofermentative and facultative heterofermentative LAB improves the performance of dairy cows.     

The effect of Lactobacillus buchneri 40788 or Lactobacillus plantarum MTD-1 on the fermentation and aerobic stability of corn silages ensiled at two dry matter contents

Whole-plant corn was harvested at 33 (normal) and 41% (moderately high) dry matter (DM) and ensiled in quadruplicate 20-L laboratory silos to investigate the effects of Lactobacillus buchneri 40788 (LB) or L. plantarum MTD-1 (LP) alone, or in combination, on the fermentation and aerobic stability of the resulting silage. Aerobic stability was defined as the amount of time after exposure to air for the silage temperature to reach 2°C above ambient temperature. The chopped forage was used in a 2 × 2 × 2 factorial arrangement of treatments: normal and moderately high DM contents, LB at 0 (untreated) or 4 × 10⁵ cfu/g of fresh forage, and LP at 0 or 1 × 10⁵ cfu/g. After 240 d of ensiling, corn silage harvested at the moderately high DM had higher pH, higher concentrations of ethanol, and more yeasts compared with the silage ensiled at the normal DM content. Inoculation with LB did not affect the concentration of lactic acid in silages with a moderately high DM, but decreased the concentration of lactic acid in the silage with normal DM. Higher concentrations of acetic acid were found in the silage treated with LB compared with those not treated with this organism. Inoculation with LP increased the concentration of lactic acid only in the silage with the normal DM content. The concentration of acetic acid was lower in silage treated with LP with a moderately high DM content, but greater in the silage treated with LP with the normal DM content when compared with silages without this inoculant. Appreciable amounts of 1,2-propanediol (average 1.65%, DM basis) were found in all silages treated with LB regardless of the DM content. The addition of L. buchneri increased the concentration of NH₃-N in silages but the addition of L. plantarum decreased it. Aerobic stability was improved in all silages treated with LB, with greater aerobic stability occurring in the silage with moderately high DM compared with silage with normal DM content. Inoculation with LP had no effect on aerobic stability. There were no interactions between L. buchneri and L. plantarum for most fermentation products or aerobic stability of the silages. This study showed that inoculating whole-plant corn with L. buchneri 40788 or L. plantarum MTD-1 has different beneficial effects on the resulting silage. There appear to be no major interactions between these organisms when added together to forage. Thus, there is potential to add both organisms simultaneously to improve the fermentation and aerobic stability of corn silage.     

Effect of Lactobacillus buchneri LN4637 and Lactobacillus buchneri LN40177 on the aerobic stability, fermentation products, and microbial populations of corn silage under farm conditions

This study determined the efficacy of the use of 2 commercial inoculants containing Lactobacillus buchneri alone or in combination with homofermentative lactic acid bacteria in improving aerobic stability of corn silage stored in commercial farm silos in northern Italy. In the first survey, samples were collected from 10 farms that did not inoculate their silages and from 10 farms that applied a Pioneer 11A44 inoculant (L. buchneri strain LN4637; Pioneer Hi-Bred International, Des Moines, IA). In the second survey, corn silage samples were collected from 11 farms that did not inoculate their silages and from 11 farms that applied a Pioneer 11CFT inoculant (L. buchneri strain LN40177; Pioneer Hi-Bred International). Inoculants were applied directly through self-propelled forage harvesters, at the recommended rate of 1 g/t of fresh forage, to achieve a final application rate of 1.0 × 10⁵ cfu/g of L. buchneri. One corn bunker silo, which had been open for at least 10 d, was examined in detail on each farm. The silages inoculated with L. buchneri had lower concentrations of lactic acid, a lower lactic-to-acetic acid ratio, a lower yeast count, and higher aerobic stability compared with the untreated silages. Unexpectedly, concentrations of acetic acid and 1,2-propanediol, 2 hallmarks of L. buchneri activity, did not differ between treatments and were only numerically higher in the inoculated silages compared with untreated ones, in both surveys. Aerobic stability, on average, was 107 and 121 h in the inoculated silages and 64 and 74 h in the untreated silages, for surveys 1 and 2, respectively, and decreased exponentially as the yeast count in the silage at the time of sampling increased, regardless of treatment. Inoculation with L. buchneri proved to be effective in reducing the yeast count to <2 log cfu/g of silage in 16 of 21 of the studied farm silages, confirming the ability of this inoculum to enhance the aerobic stability of corn silages in farm bunker silos.     

The development of lactic acid bacteria and Lactobacillus buchneri and their effects on the fermentation of alfalfa silage

This study was conducted to document the development of populations of lactic acid bacteria (LAB) and Lactobacillus buchneri in alfalfa silage treated with various inoculants. Wilted and chopped alfalfa (45% dry matter) was treated with 1) distilled water (untreated, U), 2) Lactobacillus buchneri 40788 (4 × 10⁵ cfu/g; LB), or 3) L. buchneri 40788 (4 × 10⁵ cfu/g) and Pediococcus pentosaceus (1 × 10⁵ cfu/g; LBPP). Forages were packed into triplicate vacuum-sealed, nylon-polyethylene bags per treatment, and ensiled for 2, 5, 45, 90, and 180 d. Viable (cfu) LAB in forage and silage were quantified by traditional plating on selective agar, and numbers of L. buchneri (cfu-equivalent, cfu-E) were quantified by real-time quantitative PCR. Fresh, untreated forage had 5.52 log cfu of LAB/g and 3.79 log cfu-E of L. buchneri/g. After 2 d of ensiling, numbers of LAB increased to >8 log cfu/g in all silages. In contrast, numbers of L. buchneri in U remained below 4 log cfu-E/g but reached approximately 7 log cfu-E/g in LB and LBPP. From d 5 onward, numbers of L. buchneri in U remained below 6 log cfu-E/g but approached 9 log cfu-E/g in LB and LBPP. The pH was lower in LBPP compared with U and LB after 2 and 5 d of ensiling, but pH was lower for U compared with LB and LBPP thereafter. Treatments LB and LBPP had more acetic acid than U at 45 d of ensiling, which coincided with detectable amounts of 1,2 propanediol. Inoculation with LBPP resulted in silage with the highest concentration of 1,2 propanediol after 180 d of ensiling. From d 45 onward, LB and LBPP silages had lower concentrations of residual water-soluble carbohydrates but had higher concentrations of ammonia-N than U. In conclusion, epiphytic L. buchneri can be detected in alfalfa but this population is unable to lead the silage fermentation. In contrast, when L. buchneri was added to silage as an inoculant, the numbers of L. buchneri (cfu-E) increased markedly but did not dictate fermentation until 45 d of ensiling. These findings help to explain why the response (in increased acetic acid) from the addition of L. buchneri in silages is not immediate.     

The effect of Lactobacillus buchneri and other additives on the fermentation and aerobic stability of barley silage

Whole-plant barley (39.4% dry matter) was treated with various chemical and biological additives to assess their effects on silage fermentation and aerobic stability. Treatments were untreated forage, forage treated with several amounts of Lactobacillus buchneri and enzymes (L. buchneri at 1 x 10(5), 5 x 10(5), and 1 x 10(6) cfu/g of fresh forage), forage treated with an inoculant containing (Lactobacillus plantarum, Pediococcus pentosaceus, Propionibacterium freudenreichii, and enzymes), or forage treated with a buffered propionic acid-based additive (0.2% of fresh weight). Sixty-nine d after ensiling, silages treated with L. buchneri and enzymes had lower pH, but had higher concentrations of acetic and propionic acids and higher concentrations of ethanol when compared with untreated silage. Silage treated with the multistrain inoculant containing L. plantarum had lower pH and higher concentrations of lactic acid, but lower concentrations of ammonia-N, neutral detergent fiber, and acid detergent fiber than did untreated silage. The addition of the buffered propionic acid additive resulted in silage with higher concentrations of lactic and acetic acid compared with untreated silage. Numbers of yeasts in all silages were low at silo opening (less than 3.0 log cfu/g) and were numerically the lowest in silages treated with L. buchneri but only treatment with the intermediate and high level of L. buchneri improved the aerobic stability of silage. Because of the altered fermentation pattern, inoculation with L. buchneri, when applied at equal to or more than 5 x 10(5) cfu/g, and enzymes improved the aerobic stability of barley silage.     

A meta-analysis of the effects of Lactobacillus buchneri on the fermentation and aerobic stability of corn and grass and small-grain silages

The results of adding Lactobacillus buchneri to silages from 43 experiments in 23 sources reporting standard errors were summarized using meta-analysis. The effects of inoculation were summarized by type of crop (corn or grass and small grains) and the treatments were classified into the following categories: 1) untreated silage with nothing applied (LB0), 2) silage treated with L. buchneri at ≤100,000 cfu/g of fresh forage (LB1), and 3) silage treated with L. buchneri at > 100,000 cfu/g (LB2). In both types of crops, inoculation with L. buchneri decreased concentrations of lactic acid, and this response was dose-dependent in corn but not in grass and small-grain silages. Treatment with L. buchneri markedly increased the concentrations of acetic acid in both crops in a dose-dependent manner. The numbers of yeasts were lower in silages treated with LB1 and further decreased in silages treated with LB2 compared with untreated silages. Untreated corn silage spoiled after 25 h of exposure to air but corn silage treated with LB1 did not spoil until 35 h, and this stability was further enhanced to 503 h with LB2. In grass and small-grain silages, yeasts were nearly undetectable; however, inoculation improved aerobic stability in a dose-dependent manner (206, 226, and 245 h for LB0, LB1, and LB2, respectively). The recovery of DM after ensiling was lower for LB2 (94.5%) when compared with LB0 (95.5%) in corn silage and was lower for both LB1 (94.8%) and LB2 (95.3%) when compared with LB0 (96.6%) in grass and small-grain silages.     

The effect of Lactobacillus buchneri 40788 on the fermentation and aerobic stability of ground and whole high-moisture corn

Experiments were conducted to evaluate the effects of inoculating high-moisture corn (HMC) with Lactobacillus buchneri40788 on silage fermentation and aerobic stability. In the first experiment, HMC (73% DM) was ground and treated with nothing, L. buchneri40788 to achieve 6.6 × 10⁵ cfu/g of HMC (LB), a mixture of enzymes (ENZ), LB + ENZ, or 0.1% (wet weight basis) of a liquid mold inhibitor and was ensiled in 20-L bucket silos for 90 d. Treatments with LB and LB + ENZ increased the concentrations of acetic acid and improved the aerobic stability of ground HMC relative to other treatments. Treatment ENZ had no effect on the chemical composition or aerobic stability of ground HMC. The only effect of the liquid mold inhibitor relative to untreated HMC was that it increased the concentration of propionic acid, but this did not improve its aerobic stability. In a second experiment, HMC (75% DM) was harvested as the intact, whole grain and treated with nothing, L. buchneri40788 to achieve 4 × 10⁵ cfu/g of HMC, L. buchneri40788 to achieve 6 × 10⁵ cfu/g of HMC, or L. buchneri40788 to achieve 8 × 10⁵ cfu/g of HMC and ensiled for 120 d. Treatments with L. buchneri40788 resulted in whole HMC with lower concentrations of water-soluble carbohydrates; higher concentrations of lactic, acetic, and propionic acids; and greater numbers of lactic acid bacteria but fewer molds when compared with untreated corn. As a group, inoculated silages were more aerobically stable than untreated silage, but increasing levels of application did not further improve the response. These experiments showed that addition of L. buchneri40788, but not addition of an enzyme mixture or a liquid mold inhibitor, improved the aerobic stability of ground and whole HMC harvested between 73 and 75% DM.     

Ensiling characteristics and aerobic stability of direct‐cut and wilted grass silages inoculated with Lactobacillus casei or Lactobacillus buchneri

Two experiments were conducted to study the effects of wilting and inoculating Lactobacillus casei or Lactobacillus buchneri on the fermentation and aerobic stability of grass silages. Chopped Italian ryegrass (IR) and Festulolium (FE) were ensiled with or without wilting and added L casei (>10⁶ cfu g^?1 fresh matter (FM)) or L buchneri (>10⁶ cfu g^?1 FM). Silos were opened after 120 days and microbial counts, fermentation products and aerobic stability were determined. Addition of L casei increased lactic acid and decreased acetic acid and dry matter loss regardless of wilting and forage species. Inoculation of L buchneri decreased lactic acid and increased acetic acid and 1,2‐propanediol, while the effects appeared greater in direct‐cut than in wilted grass silages. Although 1,2‐propanediol accumulated in FE silage, the diol was degraded further to propionic acid and 1‐propanol in IR silage. The activity of 1,2‐propanediol degradation was lowered when IR was wilted prior to ensiling; 1,2‐propanediol remained and the production of propionic acid and 1‐propanol was less than one‐third of that in direct‐cut silage. Regardless of forage species, addition of L buchneri increased dry matter loss compared with the untreated control, whereas the loss was not significant in wilted silages. Ammonia production was increased by L buchneri in direct‐cut but not in wilted silages. No heating was observed with or without inoculation in direct‐cut IR silage after exposure to air. Other silages were deteriorated when L buchneri was not inoculated, while the spoilage was accelerated when L casei was added to FE. Copyright © 2005 Society of Chemical Industry     

Aerobic deterioration stimulates outgrowth of spore-forming Paenibacillus in corn silage stored under oxygen-barrier or polyethylene films

The occurrence of Bacillus and Paenibacillus spores in silage is of great concern to dairy producers because their spores can survive pasteurization and some strains are capable of subsequently germinating and growing under refrigerated conditions in pasteurized milk. The objectives of this study were to verify the role of aerobic deterioration of corn silage on the proliferation of Paenibacillus spores and to evaluate the efficacy of oxygen-barrier films used to cover silage during fermentation and storage to mitigate these undesirable bacterial outbreaks. The trial was carried out on whole-crop maize (Zea mays L.) inoculated with a mixture of Lactobacillus buchneri, Lactobacillus plantarum, and Enterococcus faecium. A standard polyethylene film and a polyethylene-polyamide film with an enhanced oxygen barrier were used to produce the silage bags for this experiment. The silos were stored indoors at ambient temperature (18 to 22°C) and opened after 110 d. The silage was sampled after 0, 2, 5, 7, 9, and 14 d of aerobic exposure to quantify the growth of endospore-forming bacteria during the exposure of silages to air. Paenibacillus macerans (gram-positive, facultatively anaerobic bacteria) was able to develop during the aerobic exposure of corn silage. This species was present in the herbage at harvesting, together with clostridial spores, and survived ensiling fermentation; it constituted more than 60% of the anaerobic spore formers at silage opening. During silage spoilage, the spore concentration of P. macerans increased to values greater than 7.0 log₁₀ cfu/g of silage. The use of different plastic films to seal silages affected the growth of P. macerans and the number of spores during aerobic exposure of silages. These results indicate that the number of Paenibacillus spores could greatly increase in silage after exposure to air, and that oxygen-barrier films could help to reduce the potential for silage contamination of this important group of milk spoilage microorganisms by delaying the onset of aerobic deterioration.     

Effect of applying inoculants with heterolactic or homolactic and heterolactic bacteria on the fermentation and quality of corn silage

This study examined the effect of applying different bacterial inoculants on the fermentation and quality of corn silage. Corn plants were harvested at 35% DM, chopped, and ensiled in 20-L mini silos after application of (1) deionized water (CON) or inoculants containing (2) 1 × 10⁵ cfu/g of Pediococcus pentosaceus 12455 and Propionibacteria freudenreichii (B2); (3) 4 × 10⁵ cfu/g of Lactobacillus buchneri 40788 (BUC); or (4) 1 × 10⁵ cfu/g of Pediococcus pentosaceus 12455 and 4 × 10⁵ cfu/g of L. buchneri 40788 (B500). Four replicates of each treatment were weighed into polyethylene bags within 20-L mini silos. Silos were stored for 575 d at ambient temperature (25°C) in a covered barn. After silos were opened, aerobic stability, chemical composition, and yeast and mold counts were determined. The DNA in treated and untreated silages was extracted using lysozyme/sodium dodecyl sulfate lysis and phenol/chloroform and used as a template for a conventional PCR with primers designed on the 16S rRNA gene to detect the presence of L. buchneri in all silage samples. Acetic acid concentration was greater in B2 silages versus others (6.46 vs. 4.23% DM). Silages treated with BUC and B500 had lower pH and propionic acid concentration and greater lactic acid concentration than others. The B500 silage had the greatest lactic:acetic acid ratio (1.54 vs. 0.41), and only treatment with BUC reduced DM losses (5.0 vs. 14.3%). Yeast and mold counts were less than the threshold (10⁵) typically associated with silage spoilage and did not differ among treatments. Consequently, all silages were very stable (>250 h). Aerobic stability was not improved by any inoculant but was lower in B500 silages versus others (276 vs. 386 h). The conventional PCR confirmed the presence of similar populations of L. buchneri in all silages. This may have contributed to the prolonged aerobic stability of all silages.     

Enhanced lactic acid fermentation of silage by the addition of green tea waste

The effects of green tea waste (GTW) addition on the ensiling of forage were investigated. Wet and dried GTW added at 10, 50, 100 and 200 g kg^?1 of fresh matter (FM) and at 2, 10 and 20 g kg^?1 FM, respectively, decreased pH and increased lactic acid concentration of the silages, whereas the butyric acid concentration and ammonia nitrogen content, as a proportion of a total nitrogen, were lowered, compared with silage without additives (control). To investigate the effect of GTW‐associated LAB on silage fermentation, wet GTW was sterilized by autoclaving or gamma irradiation and added at 50 g kg^?1 FM. The silages made with sterilized GTW showed higher lactic acid concentrations, and lower pH and butyric acid concentrations than controls. The counts of lactic acid bacteria (LAB) were higher in silages made with sterilized GTW than control until 10 days after ensiling. The enhanced lactic acid fermentation was not found when green tea polyphenols (GTP) were added. These data suggested that GTW could enhance LAB growth and lactic acid production of silage, particularly when added at 50 g kg^?1 FM in a wet form or at the equivalent in a dry form. Although neither GTW‐associated LAB nor GTP accounted for the enhancement of lactic acid fermentation, GTW would possibly supply some nutrients which are heat‐stable and effective for LAB growth during silage fermentation. Copyright © 2004 Society of Chemical Industry     

Meta-analysis of effects of inoculation with Lactobacillus buchneri,with or without other bacteria,on silage fermentation,aerobic stability,and performance of dairy cows

A meta-analysis of 158 peer-reviewed articles was conducted to examine effects of inoculation with Lactobacillus buchneri (LB)-based inoculants (LBB) that did or did not include homolactic or obligate heterolactic bacteria on silage fermentation and aerobic stability. A complementary meta-analysis of 12 articles examined LBB inoculation effects on dairy cow performance. Raw mean differences between inoculant and control treatment means weighted by inverse variance were compared with a hierarchical effects model that included robust variance estimation. Meta-regression and subgrouping analysis were used to identify effects of covariates including forage type, application rate (≤10⁴, 10⁵, 10⁶, or ≥ 10⁷ cfu/g as fed), bacteria type (LB vs. LB plus other bacteria), enzyme inclusion, ensiling duration, and silo type (laboratory or farm scale). Inoculation with LBB increased acetate (62%), 1, 2 propanediol (364%) and propionate (30%) concentration and aerobic stability (73.8%) and reduced lactate concentration (7.2%), yeast counts (7-fold) and mold counts (3-fold). Feeding inoculated silage did not affect milk yield, dry matter intake, and feed efficiency in lactating dairy cows. However, forage type, inoculant composition, and dose effects on silage quality measures were evident. Inoculation with LBB increased aerobic stability of all silages except tropical grasses. Adding obligate homolactic or facultative heterolactic bacteria to LB prevented the small increase in DM losses caused by LB alone. The 10⁵ and 10⁶ cfu/g rates were most effective at minimizing DM losses while aerobic stability was only increased with 10⁵, 10⁶, and ≥ 10⁷ cfu/g rates. Inoculation with LBB increased acetate concentration, reduced yeast counts and improved aerobic stability but did not improve dairy cow performance.