Inoculant effects on alfalfa silage: fermentation products and nutritive value

The effect of 14 microbial inoculants on the fermentation and nutritive value of alfalfa silages was studied under laboratory conditions. The first cut (477 g of dry matter/kg) and second cut (393 g of dry matter/kg) of a second-year alfalfa stand were ensiled in 2 trials. In both trials alfalfa was harvested with standard field equipment. All inoculants were applied at 1.0 × 10⁶ cfu/g of crop. Uninoculated silages served as controls. After inoculants were added, the chopped forages were ensiled in 1.0- and 0.5-L anaerobic glass jars, respectively, at a density of 500 g/L. Each trial had 15 treatments (uninoculated control and 14 inoculants), with 4 silos per treatment. Silos were stored for a minimum of 30 d at room temperature (∼22°C). In first-cut silage, all inoculants but one reduced pH relative to the uninoculated control, and all but 2 of the homofermentative strains shifted fermentation toward lactic acid. In second-cut silage, the epiphytic lactic acid bacterial population was 2.7 × 10⁷ cfu/g, and only commercial inoculants produced significant shifts in fermentation. Overall, microbial inoculants generally had a positive effect on alfalfa silage characteristics in terms of lower pH and shifting fermentation toward lactic acid with homofermentative lactic acid bacteria or toward acetic acid with heterofermentative lactic acid bacteria, Lactobacillus buchneri. These effects were stronger in the commercial products tested. In spite of the positive effects on silage fermentation, 48-h in vitro true DM digestibility was not improved by inoculation with lactic acid bacteria.     

Effects of tallow in diets based on corn silage or alfalfa silage on digestion and nutrient use by lactating dairy cows

Six multiparous Holstein cows (average 31 days in milk; 36.3 kg/d of milk) fitted with ruminal cannulas were used in a 6 x 6 Latin square with 21-d periods to investigate the effects of diets that varied in forage source and amount of supplemental tallow. Isonitrogenous diets in a 2 x 3 factorial arrangement were based on either high corn silage (40:10 corn silage to alfalfa silage, % of dry matter) or high alfalfa silage (10:40 corn silage to alfalfa silage, % of dry matter) and contained 0, 2, or 4% tallow. Intakes of dry matter and total fatty acids were lower when cows were fed the high corn silage diet. Tallow supplementation linearly decreased dry matter intake. Milk yield was unaffected by diet; yields of milk fat and 3.5% fat-corrected milk were higher for the high alfalfa silage diet but were unaffected by tallow. Milk fat percentage was higher for the high alfalfa silage and tended to decrease when tallow was added to the high corn silage diet. Contents of trans-C18:1 isomers in milk fat were increased by high corn silage and tallow, and tended to be increased more when tallow was fed in the high corn silage diet. Ruminal pH and acetate:propionate were lower when high corn silage was fed. Ruminal acetate:propionate decreased linearly as tallow increased; the molar proportion of acetate was decreased more when tallow was added to the high corn silage diet. Ruminal liquid dilution rates were higher for the alfalfa silage diet; ruminal volume and solid passage rates were similar among diets. Total tract apparent digestibilities of dry matter, organic matter, crude protein, starch, energy, and total fatty acids were unaffected by diet. Digestibilities of neutral detergent fiber, acid detergent fiber, hemicellulose, and cellulose were lower when high corn silage was fed. The high alfalfa silage diet increased intakes of metabolizable energy and N, and increased milk energy and productive N. Tallow decreased the amount of N absorbed but had few other effects on utilization of energy or N. Tallow linearly increased concentrations of nonesterified fatty acids and cholesterol in plasma; cholesterol was increased by high alfalfa silage. Overall, forage source had more pronounced effects on production and metabolism than did tallow supplementation. Few interactions between forage source and tallow supplementation were detected except that ruminal fermentation and milk fat content were affected more negatively when tallow was fed in the high corn silage diet.     

烤烟中草酸、苹果酸和柠檬酸含量的分析研究

以云南、四川、河南、重庆4个烟区云烟85、K326和红大3个品种的B2L、C2L、X2L等级烟叶为材料,采用GC法测定了169个烤烟烟叶样品中草酸、苹果酸和柠檬酸的含量,并对不同产地、品种、等级和部位烤烟烟叶的多元有机酸含量进行了统计分析,结果表明:①同一地区、K326和云烟85的相同等级烤烟烟叶中草酸、苹果酸和柠檬酸含量均没有显著差异;红大和云烟85的苹果酸含量没有显著差异,而红大的草酸和柠檬酸含量均高于云烟85;②不同地区、相同品种和等级的烤烟叶中,草酸含量均没有显著性差异;苹果酸、柠檬酸含量差异显著,含量的变化趋势为:河南重庆云南和四川;③同一地区和品种、不同部位的烤烟叶中,草酸含量均为上部明显高于中部和下部,而中部和下部相当;苹果酸含量在3个部位均有较大差别,依次为:上部中部下部;柠檬酸含量下部最高,中部与上部无显著差异。     

Effect of chestnut tannin on fermentation quality, proteolysis, and protein rumen degradability of alfalfa silage

Two experiments were conducted on alfalfa to investigate the effects of the addition of commercial chestnut hydrolyzable tannin at ensiling on 1) silage fermentation quality in lab-scale silos and protein degradation in the rumen, and 2) silage fermentation quality and proteolysis in bale silages. Wilted alfalfa was prepared with 4 tannin levels (0, 2, 4, and 6% on a dry matter (DM) basis; T0, T1, T2, T3, respectively) and ensiled in lab-scale silos. Silages (33% DM) were analyzed for fermentation quality, protein rumen degradability in situ, and organic matter digestibility in vitro through gas production after 120 d of conservation. Wilted alfalfa containing 0 and 4% tannin (T0 and T2) was harvested at 40% DM (wilting level I) and 53% DM (wilting level II) for bale (600 mm diameter) silage. Silages were analyzed for fermentation quality after 78 d of conservation. All the silages were well fermented with no butyric acid. Lab-scale silages showed reductions in ammonia, nonprotein nitrogen (NPN) and DM losses in T2 and T3 treatments, while the fermentation acid profiles were unaffected. In experiment 1, the untreated silage (T0) had the highest protein degradability after being incubated in the rumen. The addition of tannin reduced crude protein ruminal disappearance in a dose-dependent manner. However, the tannin reduced the organic matter digestibility by 5.1% for all of the tannin addition levels. The tannin positively affected the silage quality in the round bale silages, in particular reducing ammonia and NPN in the lowest wilting level. In both experiments, T2 treatment reduced proteolysis without any influence of DM on the binding reaction and reduced the NPN by 15% in comparison to the control.     

Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage

Four Holstein cows fitted with ruminal cannulas were used in a 4 × 4 Latin square design (28-d periods) with a 2 × 2 factorial arrangement of treatments to investigate the effects of addition of a specific mixture of essential oil compounds (MEO; 0 vs. 750 mg/d) and silage source [alfalfa silage (AS) vs. corn silage (CS)] on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition. Total mixed rations containing either AS or CS as the sole forage source were balanced to be isocaloric and isonitrogenous. In general, no interactions between MEO addition and silage source were observed. Except for ruminal pH and milk lactose content, which were increased by MEO supplementation, no changes attributable to the administration of MEO were observed for feed intake, nutrient digestibility, end-products of ruminal fermentation, microbial counts, and milk performance. Dry matter intake and milk production were not affected by replacing AS with CS in the diet. However, cows fed CS-based diets produced milk with lower fat and higher protein and urea N concentrations than cows fed AS-based diets. Replacing AS with CS increased the concentration of NH₃-N and reduced the acetate-to-propionate ratio in ruminal fluid. Total viable bacteria, cellulolytic bacteria, and protozoa were not influenced by MEO supplementation, but the total viable bacteria count was higher with CS- than with AS-based diets. The apparent digestibility of crude protein did not differ between the AS and CS treatments, but digestibilities of neutral detergent fiber and acid detergent fiber were lower when cows were fed CS-based diets than when they were fed AS-based diets. Duodenal bacterial N flow, estimated using urinary purine derivatives and the amount of N retained, increased in cows fed CS-based diets compared with those fed AS-based diets. Feeding cows AS increased the milk fat contents of cis-9, trans-11 18:2 (conjugated linoleic acid) and 18:3 (n-3 fatty acid) compared with feeding cows CS. Results from this study showed limited effects of MEO supplementation on nutrient utilization, ruminal fermentation, and milk performance when cows were fed diets containing either AS or CS as the sole forage source.     

Ryegrass or alfalfa silage as the dietary forage for lactating dairy cows

Renewed interest exists in using grass forages to dilute the higher crude protein (CP) and lower digestible fiber present in legumes fed to lactating dairy cows. A 3 x 3 Latin square feeding study with 4-wk periods was conducted with 24 Holstein cows to compare ryegrass silage, either untreated control or macerated (intensively conditioned) before ensiling, with alfalfa silage as the sole dietary forage. Ryegrass silages averaged [dry matter (DM) basis] 18.4% CP, 50% neutral detergent fiber (NDF), and 10% indigestible acid detergent fiber (ADF) (control) and 16.6% CP, 51% NDF, and 12% indigestible ADF (macerated). Alfalfa silage was higher in CP (21.6%) and lower in NDF (44%) but higher in indigestible ADF (26%). A lower proportion of the total N in macerated ryegrass silage was present as nonprotein N than in control ryegrass and alfalfa silages. Diets were formulated to contain 41% DM from either rye-grass silage, or 51% DM from alfalfa silage, plus high moisture corn, and protein concentrates. Diets averaged 17.5% CP and 28 to 29% NDF. The shortfall in CP on ryegrass was made up by feeding 7.6% more soybean meal. Intake and milk yields were similar on control and macerated ryegrass; however, DM intake was 8.3 kg/d greater on the alfalfa diet. Moreover, feeding the alfalfa diet increased BW gain (0.48 kg/d) and yield of milk (6.1 kg/d), FCM (6.8 kg/d), fat (0.26 kg/d), protein (0.25 kg/d), lactose (0.35 kg/d), and SNF (0.65 kg/d) versus the mean of the two ryegrass diets. Both DM efficiency (milk/DM intake) and N efficiency (milk-N/N-intake) were 27% greater, and apparent digestibility was 16% greater for DM and 53% greater for NDF and ADF, on the ryegrass diets. However, apparent digestibility of digestible ADF was greater on alfalfa (96%) than on ryegrass (average = 91%). Also, dietary energy content (estimated as net energy of lactation required for maintenance, milk yield, and weight gain) per unit of digested DM was similar for all three diets. Results of this trial indicated that, relative to ryegrass silage, feeding alfalfa silage stimulated much greater feed intake, which supported greater milk production.     

氨基酸对出芽短梗霉发酵生产聚苹果酸的影响

本文研究了添加氨基酸对出芽短梗霉A.pullulans CGMCC3337发酵生产聚苹果酸(PMLA)的影响。通过单因素实验和正交实验分别对氨基酸种类及其添加量进行优化。由单因素实验可知:天冬氨酸(Asp)、亮氨酸(Leu)、缬氨酸(Val)和苏氨酸(Thr)对菌体生长和PMLA合成最有利。由正交实验可知:氨基酸的最优组合为(g/L):天冬氨酸(Asp)0.4、亮氨酸(Leu)0.4、缬氨酸(Val)0.4、苏氨酸(Thr)0.4。在最优组合条件下获得的PMLA产量和分子量分别达到了57.89 g/L和8879 u,比未添加氨基酸获得的PMLA产量和分子量分别提高了40.92%和45.20%。      

Effects of a mixture of lactic acid bacteria applied as a freeze-dried or fresh culture on the fermentation of alfalfa silage

Alfalfa (approximately 31% DM) was untreated or treated with a silage inoculant containing the lactic acid bacteria Lactobacillus lactis, Lactobacillus plantarum L-54, and L. plantarum Aber F1. The inoculant was added at a normal and a high dose as a freeze-dried powder that had been mixed with water just prior to application, or it was grown with nutrients the day before and added as a fresh culture. The actual application rate of lactic acid bacteria was 1.19 × 10⁵ for the normal dose, 4.30 × 10⁵ for the high dose, and 5.10 × 10⁵ for the fresh culture. All inoculated silages showed a faster increase in the rate of lactic acid production and a decrease in the drop in pH over the first 24 h of ensiling compared with untreated silage. The effect was greatest for silage treated with the fresh culture and was supported by the fact that this treatment had numbers of lactic acid bacteria that increased faster than in other treatments. Inoculation also generally resulted in a fermentation profile that was more homolactic (more lactic acid and less acetic acid, ethanol, and NH₃-N) than for untreated silage, but the effect was greatest for the fresh culture. Inoculation did not affect in vitro neutral detergent fiber digestion or the concentrations of neutral detergent fiber or total N in silages. The recovery of dry matter was greater in silage that was treated with a high level of the freeze-dried culture or with the fresh culture when compared with the untreated control. This study showed that application of a silage inoculant as a freeze-dried culture or as a fresh culture resulted in alfalfa silage with a more homolactic fermentation profile. The effect was greatest from addition of the fresh culture.     

Effects of inoculation of high dry matter alfalfa silage on ensiling characteristics,ruminal nutrient degradability and dairy cow performance

The objective of this study was to determine the effects of a homolactic acid inoculant on ensiling characteristics and nutritive value of high dry matter (DM) alfalfa. The ensiling characteristics were determined by ensiling inoculated and untreated alfalfa haylage in mini‐silos for 0, 2, 4, 8, 16 and 45 days. Two lactating cows fitted with ruminal fistulas were used to determine ruminal degradabilities of nutrient in inoculated and untreated alfalfa silage (45‐day silage). Effects of feeding inoculated and untreated alfalfa silage on animal performance were determined using 28 lactating dairy cows fed total mixed diets (40% forage and 60% concentrate) with the forage portion consisting of inoculated or untreated alfalfa silage. The pH of the inoculated silage declined from 5.9 to 4.5 within 2 days of ensiling while the pH of the untreated silage did not drop below 5 until day 16 post‐ensiling. At all ensiling times, inoculated alfalfa silage had lower pH than untreated alfalfa silage. The concentration of lactic acid was higher while that of water‐soluble carbohydrates was lower for inoculated than untreated alfalfa silage at all ensiling times. Inoculation increased proteolysis as indicated by a reduction in true protein and neutral detergent insoluble protein and an increase in non‐protein nitrogen. Ruminal degradability of DM, crude protein and neutral detergent fiber of alfalfa silage were not affected by inoculation. The average effective degradability of dry matter, crude protein and neutral detergent fiber was 66, 65 and 62%, respectively. Dairy cows fed inoculated alfalfa silage had DM intake (average 22 kg day^?1) and milk yield (average 42 kg day^?1) similar to cows fed untreated alfalfa silage. It was concluded that the inoculant used in this study improved the ensiling characteristics of alfalfa silage with no significant effects on dairy cow performance. Copyright © 2004 Society of Chemical Industry     

Inoculant effects on alfalfa silage: in vitro gas and volatile fatty acid production

Alfalfa silages from 2 similar trials were analyzed for in vitro ruminal gas production. In both trials, there were 15 treatments: alfalfa treated at ensiling with 1 of 14 lactic acid bacterial inoculants or untreated alfalfa. First-cut (477 g of dry matter/kg) and second-cut (393 g of dry matter/kg) alfalfa were ensiled in glass jars for a minimum of 35 d at room temperature (∼22°C). At opening, a portion of each silage was wet-ground with a mixer. Each silage was then assessed for in vitro ruminal gas production in 3 replicate runs with the wet-ground silage, 1 on the fresh silage and 2 on frozen and thawed silage. In vitro gas production was measured in 160-mL sealed serum vials incubated at 39°C. One gram of silage was incubated with 17.1 mL of nutrient solution, 0.9 mL of reducing solution, and 12 mL of ruminal inoculum (1:2 vol.vol mixture of rumen fluid and buffer). Gas production was measured manually by using a pressure gauge at 3, 6, 9, 24, 48, and 96 h. At 96 h, the rumen fluid was analyzed for pH and volatile fatty acids. In the 2 trials, the untreated control silage produced either numerically the highest or one of the highest levels of gas production per unit of dry matter incubated. In first-cut silage, 9 of the inoculant treatments at 9 h and 4 treatments at 96 h had reduced gas production compared with the control. In second-cut silage, 10 inoculant treatments at both 9 and 96 h had reduced gas production compared with the control. Furthermore, in first-cut silage, the fraction of total gas production at 3, 6, and 9 h was numerically the highest for the control, and only 4 treatments were not significantly lower than the control at 9 h. In second-cut silage, 2 of 14 inoculated treatments produced faster fractional rates of gas production than the control, but most inoculated treatments had numerically slower fractional rates (4 significant) in the first 9 h. The in vitro fermented wet-ground control silages had one of the highest acetate:propionate ratios in both trials, significantly higher than 12 and 8 of the inoculated treatments in first- and second-cut silage, respectively. The response in acetate:propionate ratio in both cuts was similar, even though the control silage was highest in lactic acid in one trial and lowest in the other. Overall, inoculation of crops at ensiling appears to affect in vitro ruminal fermentation of wet-ground silages, even in the absence of large effects during silage fermentation.     

Use of exogenous fibrolytic enzymes to enhance in vitro fermentation of alfalfa hay and corn silage

Two in vitro experiments were performed to identify promising exogenous fibrolytic enzyme products (EFE) and optimum dose rates (DR) for improving the degradation of alfalfa hay and corn silage. The relationship between enzymatic activity and fermentation responses was examined to identify optimum formulations. In experiment 1, 5 EFE containing mainly endoglucanase and xylanase activities, with different ratios between the 2 activities, were assessed at a DR of 0.7, 1.4, and 2.1 mg/g of DM forage. Milled alfalfa hay or corn silage was incubated in an in vitro batch culture with buffer, ruminal fluid, and EFE. Gas production (GP) was measured during 24 h of incubation, and degradabilities of DM and fiber were measured after terminating the incubation at 24 h. Two (E1 and E3) EFE substantially improved GP and degradation of alfalfa hay and corn silage fiber. The optimum DR of these EFE was 1.4 mg/g of DM for both forages with improvements in NDF degradability up to 20.6% for alfalfa hay and up to 60.3% for corn silage. Whereas added activities of endoglucanase and exoglucanase were positively correlated with improvement in NDF degradability for alfalfa hay and corn silage, there was no relationship between added xylanase activity and NDF degradability. The 2 most promising EFE from experiment 1 were reevaluated in experiment 2, alone and in combination with a high xylanase EFE, to determine whether their effectiveness could be enhanced by decreasing the endoglucanase to xylanase ratio. The 2 EFE improved GP and fiber degradation in a manner similar to that observed in experiment 1, but the combination treatments resulted in no further beneficial effects. Exogenous fibrolytic enzyme products can greatly improve forage utilization, but DR and the activities supplied are critical for achieving this response. Products used with alfalfa hay and corn silage should contain high endoglucanase activity, with an ideal ratio of endoglucanse to xylanase.     

Influence of particle size on the effectiveness of the fiber in alfalfa silage

The objective was to determine the influence of alfalfa silage particle length on milk yield, milk composition, and chewing activity. Sixteen multiparous lactating Holsteins were used in each of two separate feeding trials over a 2-yr period providing two repetitions. Each trial was based on 4 x 4 Latin squares with 21-d periods. All four diets (2-yr average, dry basis) contained a basal level of 10.7% neutral detergent fiber from corn silage and 2.3% neutral detergent fiber from chopped alfalfagrass hay. One of the diets served as a low forage, low fiber control and contained only the basal forages. The other three diets contained an additional 8.6% neutral detergent fiber from coarser alfalfa silage, finely rechopped alfalfa silage or an equal mixture ofthe two. An increase in the forage content above the basal amount using alfalfa silage increased 4% fat-corrected milk yield, milk fat yield and concentration, eating time, and total chewing time. Dry matter intake was not influenced by diet. Linear increases in rumination and total chewing times were observed as the mean particle size of the alfalfa silage increased from finer to coarser. There was no linear effect of alfalfa silage particle size on milk yield, 4% fat-corrected milk yield, dry matter intake, or milk composition.     

Effect of supplemental tallow on performance of dairy cows fed diets with different corn silage:alfalfa silage ratios

A study was conducted to investigate the response to supplemental tallow of lactating cows fed basal diets with different alfalfa silage:corn silage ratios. We postulated that supplemental tallow will have decreasing negative effects on rumen fermentation, dry matter intake (DMI), and milk fat percentage as the dietary ratio of alfalfa silage:corn silage is increased. Eighteen Holstein cows averaging 134 +/- 14 d in milk were used in a replicated 6 x 6 Latin square design with 21-d periods. Treatments were arranged as a 2 x 3 factorial with 0 or 2% tallow (DM basis) and three forage treatments: 1) 50% of diet DM as corn silage, 2) 37.5% corn silage and 12.5% alfalfa silage, and 3) 25% corn silage and 25% alfalfa silage. Cows were allowed ad libitum consumption of a total mixed ration. Diets were formulated to contain 18% crude protein and 32% neutral detergent fiber. No fat x forage treatment interactions were observed. Fat supplemented cows had lower DMI and produced more milk with less milk fat content relative to non-supplemented cows. Concentration of trans-octadecenoic acids was higher in milk fat of tallow-supplemented cows. Tallow supplementation had no effect on ruminal pH and acetate:propionate ratio, but tended to decrease total volatile fatty acid (VFA) concentration in the rumen. Increasing the proportion of alfalfa silage increased DMI, milk fat percentage, and milk fat yield regardless of the fat content of the diet. Total VFA concentration and acetate:propionate ratio in the rumen were increased in response to higher levels of alfalfa in the diets. These results suggest that replacing corn silage with alfalfa silage did not alleviate the negative response of dairy cows to tallow supplementation at 2% of diet DM.     

Effects of dairy slurry on silage fermentation characteristics and nutritive value of alfalfa

Dairy producers frequently ask questions about the risks associated with applying dairy slurry to growing alfalfa (Medicago sativa L.). Our objectives were to determine the effects of applying dairy slurry on the subsequent nutritive value and fermentation characteristics of alfalfa balage. Dairy slurry was applied to 0.17-ha plots of alfalfa; applications were made to the second (HARV1) and third (HARV2) cuttings during June and July of 2012, respectively, at mean rates of 42,400 ± 5271 and 41,700 ± 2397 L/ha, respectively. Application strategies included (1) no slurry, (2) slurry applied directly to stubble immediately after the preceding harvest, (3) slurry applied after 1 wk of post-ensiled regrowth, or (4) slurry applied after 2 wk of regrowth. All harvested forage was packaged in large, rectangular bales that were ensiled as wrapped balage. Yields of DM harvested from HARV1 (2,477 kg/ha) and HARV2 (781 kg/ha) were not affected by slurry application treatment. By May 2013, all silages appeared to be well preserved, with no indication of undesirable odors characteristic of clostridial fermentations. Clostridium tyrobutyricum, which is known to negatively affect cheese production, was not detected in any forage on either a pre- or post-ensiled basis. On a pre-ensiled basis, counts for Clostridium cluster 1 were greater for slurry-applied plots than for those receiving no slurry, and this response was consistent for HARV1 (4.44 vs. 3.29 log₁₀ genomic copies/g) and HARV2 (4.99 vs. 3.88 log₁₀ genomic copies/g). Similar responses were observed on a post-ensiled basis; however, post-ensiled counts also were greater for HARV1 (5.51 vs. 5.17 log₁₀ genomic copies/g) and HARV2 (5.84 vs. 5.28 log₁₀ genomic copies/g) when slurry was applied to regrowth compared with stubble. For HARV2, counts also were greater following a 2-wk application delay compared with a 1-wk delay (6.23 vs. 5.45 log₁₀ genomic copies/g). These results suggest that the risk of clostridial fermentations in alfalfa silages is greater following applications of slurry. Based on pre- and post-ensiled clostridial counts, applications of dairy slurry on stubble are preferred (and less risky) compared with delayed applications on growing alfalfa.     

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.     

Effects of 8 chemical and bacterial additives on the quality of corn silage

This project aimed to evaluate the effects 8 additives on the fermentation, dry matter (DM) losses, nutritive value, and aerobic stability of corn silage. Corn forage harvested at 31% DM was chopped (10 mm) and treated with (1) deionized water (control); (2) Buchneri 500 (BUC; 1 × 10⁵ cfu/g of Pediococcus pentosaceus 12455 and 4 × 10⁵ cfu/g of Lactobacillus buchneri 40788; Lallemand Animal Nutrition, Milwaukee, WI); (3) sodium benzoate (BEN; 0.1% of fresh forage); (4) Silage Savor acid mixture (SAV: 0.1% of fresh forage; Kemin Industries Inc., Des Moines, IA); (5) 1 × 10⁶ cfu/g of Acetobacter pasteurianus-ATCC 9323; (6) 1 × 10⁶ cfu/g of Gluconobacter oxydans-ATCC 621; (7) Ecosyl 200T (1 × 10⁵ cfu/g of Lactobacillus plantarum MTD/1; Ecosyl Products Inc., Byron, IL); (8) Silo-King WS (1.5 × 10⁵ cfu/g of L. plantarum, P. pentosaceus and Enterococcus faecium; Agri-King, Fulton, IL); and (9) Biomax 5 (BIO; 1 × 10⁵ cfu/g of L. plantarum PA-28 and K-270; Chr. Hansen Animal Health and Nutrition, Milwaukee, WI). Treated forage was ensiled in quadruplicate in mini silos at a density of 172 kg of DM/m³ for 3 and 120 d. After 3 d of ensiling, the pH of all silages was below 4 but ethanol concentrations were least in BEN silage (2.03 vs. 3.24% DM) and lactic acid was greatest in SAV silage (2.97 vs. 2.51% DM). Among 120-d silages, additives did not affect DM recovery (mean = 89.8% ± 2.27) or in vitro DM digestibility (mean = 71.5% ± 0.63). The SAV silage had greater ammonia-N (0.85 g/kg of DM) and butyric acid (0.22 vs. 0.0% DM) than other treatments. In contrast, BEN and Silo-King silages had the least ammonia-N concentration and had no butyric acid. The BEN and A. pasteurianus silages had the lowest pH (3.69) and BEN silage had the least ethanol (1.04% DM) and ammonia nitrogen (0.64 g/kg DM) concentrations, suggesting that fermentation was more extensive and protein degradation was less in BEN silages. The BUC and BIO silages had greater acetic acid concentrations than control silages (3.19 and 3.19 vs. 2.78% DM), but yeast counts did not differ. Aerobic stability was increased by 64% by BUC (44.30 h) and by 35% by BEN (36.49 h), but other silages had similar values (27.0 ± 1.13 h).     

E. coli o157:H7 population reduction from alfalfa seeds with malic acid and thiamine dilauryl sulfate and quality evaluation of the resulting sprouts

It has been reported that washing seeds with a 20000 ppm Ca(OCl)(2) solution as recommended by the U.S. Food and Drug Administration is unable to eliminate E. coli cells attached to seed surfaces, and the bacterial cells that have survived a sanitation wash can proliferate during sprouting to a high population. The objectives of this research were to examine the efficacy of malic acid (MA) and thiamine dilauryl sulfate (TDS) combined treatments on the inactivation of E. coli O157:H7 on alfalfa seeds, to study the growth of the remaining E. coli cells during sprouting, and to evaluate the sprout quality. When 10 g of inoculated alfalfa seeds were washed in a 10% MA-1% TDS solution, a complete elimination of E. coli was achieved. The same result was observed by washing the seeds in a 20000 ppm Ca(OCl)(2) solution. However, when the seed size was increased to 50 g while maintaining the same seed-to-sanitizer ratio, both the MA + TDS and the 20000 ppm chlorine washes failed to completely inactivate the E. coli cells on the seeds. Nevertheless, the 10% MA-1% TDS solution was significantly more effective in E. coli count reduction compared to the 20000 ppm chlorine wash. The E. coli O157:H7 cells remaining on the seeds after treatments with both sanitizers grew up to 7 to 8 log CFU/g sprout after 96 h of sprouting. Under the treatment conditions used in this study, none of the treatments resulted in significant differences in germination rate, yield, or quality of the sprouts. PRACTICAL APPLICATION: The malic acid (MA) and thiamine dilauryl sulfate (TDS) combined treatment may provide a new solution to secure the microbial safety of seeds and sprouts. An important finding of this study is that seed sample size has a significant impact on the inactivation of E. coli O157:H7 on alfalfa seeds. The microbial inactivation results obtained in a laboratory set-up cannot be directly applied to a large scale operation. A validation test on the large scale has to be performed to evaluate the efficacy of the sanitizer.