Proteolysis, fermentation efficiency, and in vitro ruminal digestion of peanut stover ensiled with raw or heated corn

Peanut stover (PS) is similar to full-bloom alfalfa hay in chemical composition. The objective of this study was to assess the effect of adding raw or heated corn meal to PS at ensiling on silage N components, fermentation acids, and digestion by ruminal microorganisms. The PS was collected after harvesting of peanuts and ensiled immediately without and with addition of raw or heated corn meal (100 g/kg of fresh weight). Corn was added to PS so that the initial mixture would contain adequate dry matter (DM) (approximately 30%) and additional nonfiber carbohydrate to enhance silage fermentation. After 8 wk of silo fermentation, corn-treated silages contained less structural carbohydrates but more non-fiber carbohydrates compared with the untreated control. A shift from hemicellulose to nonfiber carbohydrate use during silage fermentation was evident by corn treatment. Additional corn at ensiling resulted in silage N with less water-soluble N, protein N, nonprotein N, nonprotein nonammonia N (peptides plus amino acids), and ammonia N. Based on changes in soluble nonprotein N before and after ensiling, the amount of proteolysis was approximately 66% for control silage and was nearly 40% lower in response to corn treatment. Adding corn increased silage lactic acid, but both acetic and propionic acids decreased. These changes were reflected in the lower pH and higher fermentation efficiency with corn-treated silages. More DM was digested and greater amounts of volatile fatty acids, except for branched-chain acids, were produced in vitro by ruminal microorganisms with corn-treated silages. In addition, incubations with silage treated with heated corn contained higher concentrations of acetic and propionic acids compared with raw corn. In vitro ammonia accumulation per unit of DM digested was lower for corn treatments than the control, and for heated corn vs. raw corn-treated silage. These results indicate that supplementation of either raw or heated corn on PS at ensiling could minimize proteolysis and improve fermentation efficiency. Advantages from using heated vs. raw corn could extend beyond silage fermentation and include rumen microbial fermentation.     

The effect of delayed ensiling and application of a propionic acid-based additive on the fermentation of barley silage

Prolonged exposure to air can adversely affect the silage fermentation process. To investigate a possible method to overcome this problem, we determined if a buffered propionic acid-based additive, applied to chopped, whole-plant barley exposed to air before ensiling, would affect the subsequent fermentation. Wilted forage was chopped and treated with nothing, or with 0.1% (wt/wt wet forage) of a buffered propionic acid-based additive and ensiled immediately in quadruplicate 20-L laboratory silos. Portions of the chopped forage, untreated and treated, were left in loose piles in a barn for 24 h before ensiling. Another portion of the untreated silage exposed to air for 24 h was also treated with 0.1% of the additive just before ensiling. Prolonged exposure to air before ensiling increased the numbers of yeasts on forages by more than 1,000-fold. The concentrations of water-soluble carbohydrates decreased by more than 50%; the ammonia-N concentrations increased 40%, and pH increased by more than 1 unit as a result of exposure to air. These changes were less in forage that was treated with the additive at chopping. After 60 d, silages of forages that were exposed to air before ensiling had a higher pH, higher concentrations of ammonia-N and butyric acid, and lower concentrations of lactic and acetic acids than silages of forage that had been ensiled immediately after harvest. In situ DM digestibility was lowest in untreated silages that had been exposed to air before ensiling. In contrast, treatment with the additive, applied before or after exposure to air, prevented the reduction in in vitro digestion.     

The effect of sorbic acid on loss reduction during storage of orange peels

Sorbic acid (SA) at 0·25, 0·5 and 1·0 g kg^?1 was applied to fresh orange peels before ensiling in special 18 litre fermentation containers in two separate experiments. The higher two concentrations of sorbic acid were effective in reducing dry matter (DM) loss throughout the ensiling period, up to 90 days. Chemical analysis revealed that SA at the higher concentrations slowed down the fermentation rate, with less water-soluble carbohydrates (WSC) converted into organic acids and ethanol after 5 days of ensiling. However, after 30 and 90 days of ensiling, similar amounts of lactic acid were found in all silages, but less ethanol and more residual WSC were found in the silage treated with the higher SA concentrations. This indicates a more efficient fermentation pattern in the SA-treated silages. Microbial examination of the fresh and ensiled peels indicated large numbers of yeasts, the activity of which was probably inhibited by the sorbic acid.     

Effects of an exogenous protease on the fermentation and nutritive value of corn silage harvested at different dry matter contents and ensiled for various lengths of time

The objective of this experiment was to evaluate the effects of adding an experimental protease to corn plants harvested at different maturities on silage fermentation and in vitro ruminal starch digestibility (IVSD). Corn plants were harvested at maturities resulting in plants with 31 or 40% dry matter (DM). Plants were chopped, kernel processed, and treated with (1) only a 0.1 M phosphate buffer (pH 5.5, 5% vol/wt of fresh forage), (2) buffer with protease to obtain a final concentration of 20 mg of protease/kg of wet forage, and (3) buffer with protease to obtain a final concentration of 2,000 mg of protease/kg of wet forage. Treated forages (about 500 g) were ensiled in nylon-polyethylene pouches and stored between 21 and 23°C for 0, 45, 90, and 150 d. Data were analyzed as a 2 × 3 × 4 factorial arrangement of treatments, with the main effects of harvest DM, dose of protease, days of ensiling, and their interactions. The treatment with the highest dose of protease resulted in more robust fermentations across harvest DM with higher concentrations of lactic and acetic acids compared with untreated silage. Concentrations of soluble protein (% of crude protein) increased with time of ensiling, regardless of DM content at harvest. However, averaged over both harvest DM contents, it increased by 37% for silages treated with the high dose of protease compared with an average 11% increase for untreated silages and silage treated with the low dose of protease, between d 0 and 45. Averaged over both harvest DM contents, the concentration of soluble protein peaked in silages treated with the high dose of protease after 45 d of ensiling, whereas it peaked at d 90 in untreated silages and silage treated with the low dose of protease. Similar changes occurred in the concentration of NH₃-N due to length of ensiling and treatment with protease. In fresh forages, the concentration of starch for early- and late-harvested forages was similar, but IVSD was lower in the latter. After 45 d of ensiling, IVSD was highest in both early- and late-harvested silages that were treated with the high level of protease. After 150 d of ensiling, IVSD was similar among silages treated with protease, regardless of DM at harvest. Treating corn plants with a high dose of an experimental protease at harvest accelerated proteolysis during ensiling, resulting in corn silages with levels of IVSD after 45 d of ensiling that were only obtained in untreated corn silages after 150 d of ensiling.     

The effect of treating alfalfa with Lactobacillus buchneri 40788 on silage fermentation,aerobic stability,and nutritive value for lactating dairy cows

Lactobacillus buchneri 40788 and enzymes (beta-glucanase, alpha-amylase, xylanase, and galactomannase) were applied to chopped alfalfa (39% DM) to study their effects on the fermentation and nutritive value of the silage. Alfalfa was treated with nothing, or L. buchneri 40788, for a final application rate of 1 x 10(5), 5 x 10(5), or 1 x 10(6) cfu/g of fresh forage and ensiled in laboratory silos for 2, 4, 8, and 56 d. Treatment with L. buchneri 40788 had few effects on the end products of fermentation through 8 d of ensiling. However, after 56 d of ensiling, treated silages had a higher pH (4.55 vs. 4.38) and higher concentrations of acetic acid (6.40 vs. 4.24%), propionic acid (0.18 vs. 0.06%), and ammonia-N (0.35 vs. 0.29%) when compared to untreated silage. Lactic acid was also numerically lower in treated (3.51%) than untreated (4.12%). Silages treated with the moderate and highest dose of L. buchneri 40788 also resulted in greater recoveries of DM than did untreated silage. Alfalfa (43% DM) was also untreated or treated with a commercial application of L. buchneri 40788 (4 x 10(5) cfu/g, a commercial dose) in farm-scale bag silo. Holstein cows were fed a diet comprised of 32% untreated or treated alfalfa silage, 11% corn silage, 5% chopped alfalfa hay, and 52% of concentrate (DMB) for a 6-wk treatment period. Dry matter intake and milk composition were unaffected by treatment, but cows fed silage treated with L. buchneri 40788 produced 0.8 kg more milk than did cows fed untreated silage. Treated silage had a higher concentration of acetic acid (5.67 vs. 3.35%) but lower lactic acid (3.50 vs. 4.39%) than untreated silage. When exposed to air, the total mixed ration containing treated alfalfa silage remained stable for 100 h, whereas the ration containing untreated silage spoiled after 68 h. Treating alfalfa silage with L. buchneri 40788 increased the concentration of acetic acid, and when the silage was combined into a total mixed ration and fed to lactating cows, it improved the aerobic stability of the ration and increased milk production.     

Inoculation with Lactobacillus plantarum of Alfalfa,Corn, Sorghum,and Wheat Silages

Alfalfa, corn, sorghum, and wheat forages were harvested for ensiling. Effects of inoculation of the forages with Lactobacillus plantarum (10⁷/g silage) were studied in small experimental 55-kg drum silos. Control and treated silages were prepared, and drums from each treatment were opened on days 1, 2, 4, 8, 16, and 33. Proximate analysis, pH, neutral detergent fiber, acid detergent fiber, soluble carbohydrate, lactic acid, and volatile fatty acid contents were measured. Total faculative anaerobic microflora were increased in all silages by addition of L. plantarum, and numbers of lactobacilli, were increased in all but corn silage. Yeasts and molds were lower only in inoculated alfalfa and wheat silages. The effect of the inoculum was observed on chemical composition only for alfalfa and wheat silages. These treated silages had a lower pH, higher lactic acid content, and great recovery of dry matter, crude protein, and detergent fiber (alfalfa) and nitrogen-free extract (wheat). Inoculation has an effect on microflora in all silages and a positive beneficial effect on nutrient recovery in alfalfa and wheat silages.     

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 mixing red clover with alfalfa at different ratios on dynamics of proteolysis and protease activities during ensiling

This study was conducted to study the effects of ensiled alfalfa (Medicago sativa) and red clover (Trifolium pratense) at different ratios on dynamics of fermentation parameters, N distribution, protein fractions, and protease activities during ensiling. Alfalfa and red clover were harvested and wilted to 35 and 25% dry matter, respectively, chopped to 1 cm, mixed, weighed into 1.0-L buckets at a density of 700 g/L, and ensiled for 1, 3, 7, 15, and 30 d at 30°C. The treatments were mixing ratio of alfalfa to red clover at 100:0, 70:30, 50:50, 30:70, and 0:100 (R0, R30, R50, R70, and R100, respectively; fresh weight). For each ensiling duration, 3 replicates of each treatment were prepared. With increasing proportion of red clover in silage, total N content and proportions of nonprotein N, peptide N, free amino acid N, and NH₃-N decreased linearly, and PC (indigestible true protein, acid detergent insoluble N) proportion increased linearly after ensiling. Moreover, the final pH was lower in R50 and R100 than R0 (4.29, 4.20 vs. 4.48, respectively) on d 30. Also, lactic acid concentration on d 30 was higher in R50, R70, and R100 silage compared with R0 (7.77, 7.66, and 8.76 vs. 6.34, % of dry matter, respectively). The proportion of NH₃-N in R50 was lower than in R0 but closer to R100 after ensiling. During ensiling, proteases including carboxypeptidase, aminopeptidase, and acid proteinase activities decreased as red clover proportion increased. However, no differences were detected in aminopeptidase and acid proteinase activities among R50, R70, and R100 during ensiling. Overall, 50:50 was the optimal mixing ratio of alfalfa with red clover, showing good fermentation quality with lower pH and higher lactic acid concentration, reduced protease activities and proteolysis compared with pure alfalfa silage, and also more total N content than pure red clover silage.     

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.     

The effects of sorbates on the ensilage of chopped whole-plant maize and lucerne

The incorporation of (a) sorbic acid (0.18 and 0.90 g kg^?1, fresh weight basis) and potassium sorbate (0.90 g kg^?1) with chopped lucerne, and (b) sorbic acid (0.90 g kg^?1) and potassium sorbate (0.90 g kg^?1) with chopped maize (whole-plant) at the time of ensiling, led to a reduction in the surface spoilage associated with the ensilage of these two forages. Use of sorbic acid (0.90 g kg^?1) and potassium sorbate (0.90 g kg^?1) led to (a) a reduction of volatile nitrogen (VN), higher levels of residual water-soluble carbohydrates (WSC) in lucerne silage and more aerobically stable lucerne silage and (b) a reduction in weight loss, population of yeasts and moulds, and VN, higher levels of WSC in maize silage and more aerobically stable maize silage when compared to the untreated forages or forages treated with sorbic acid and potssium sorbate at 0.045 and 0.18 g kg^?1 (fresh weight basis), respectively.     

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.     

An evaluation of the effectiveness of a chemical additive based on sodium benzoate,potassium sorbate,and sodium nitrite on the fermentation and aerobic stability of corn silage

We evaluated the effectiveness of an additive comprising sodium benzoate, potassium sorbate, and sodium nitrite (SSL) as active ingredients for its ability to improve the aerobic stability of corn silages made in North America. In experiment 1, treatment with SSL (1.5 and 2.0 L/t) on whole-plant corn (WPC) was compared with treatment with an additive containing buffered propionic acid and citric acid (BPA; 2 L/t) on corn harvested at 32 and 38% dry matter and ensiled for 120 d. Silage treated with BPA was higher in ammonia-N and propionic acid relative to other treatments. Treatments with all of the additives had numerically, but not statistically, fewer yeasts compared with untreated silage. Both application rates of SSL resulted in lower concentrations of ethanol compared with untreated and BPA silages. Treatment with BPA improved the aerobic stability of silages compared with untreated silage, but the effect from SSL was markedly greater. In experiment 2, WPC was untreated or treated with 2 or 3 L of SSL/t or a microbial inoculant containing Enterococcus faecium M74, Lactobacillus plantarum CH6072, and Lactobacillus buchneri LN1819 (final total lactic acid bacteria application rate of 150,000 cfu/g of fresh forage). Silages were air stressed for 24 h at 28 and 42 d of storage and ensiled for 49 d before opening. Inoculation had no effect on acid end products, ethanol, number of yeasts, or aerobic stability compared with other treatments. Treatment with SSL decreased the amount of ethanol, had no effect on number of yeasts, and improved aerobic stability in a dose-dependent manner compared with other treatments. In experiment 3, WPC was untreated or treated with 2 L of SSL/t and ensiled for 5, 15, and 30 d. Treatment with SSL resulted in silage with fewer yeasts and lower concentrations of ethanol after all times of ensiling compared with untreated silage. In addition, SSL improved aerobic stability after each period of ensiling, but the effect was more at 15 and 30 d compared with 5 d of storage. Treating WPC with SSL can improve the aerobic stability of corn silage made in North America, and the effect can be observed as soon as 5 d after ensiling.     

Comparison of grass and legume silages for milk production. 2. In vivo and in sacco evaluations of rumen function

Two experiments were conducted to investigate the basis for higher voluntary intakes and increased alpha-linolenic acid content in milk from cows offered clover silages. Six cows with rumen and duodenal cannulae were used in a four-period changeover-design experiment. Cows received 8 kg/d of dairy concentrate and had ad libitum access to one of six silage treatments: grass, red clover, white clover, alfalfa, and 50/50 (dry matter basis) mixtures of grass with red clover or white clover. The rumen fermentability of grass, red clover, white clover, and grass/red clover silages was also evaluated in a nylon bag study. Legume silages led to increased dry matter intake and milk production in comparison with grass silage. There was no significant effect of legume silages on rumen pH and volatile fatty acid concentrations, but a significant increase in rumen ammonia concentration with the legume silages, reflecting their higher protein content. The inclusion of white clover or alfalfa silage, but not red clover silage, in diets led to an increase in molar proportions of isobutyric, iso-valeric, and n-valeric acids in comparison with diets based on grass silage. Rumen fill was significantly lower, and rumen passage rates were significantly higher for cows offered alfalfa or white clover silages. However, the markedly different particle size distribution of rumen contents with these feeds suggests very different mechanisms for the high intake characteristics: high rates of particle breakdown and passage with alfalfa, and high rates of fermentation and passage with white clover. Microbial energetic efficiency (grams microbial N per kilogram organic matter apparently digested in the rumen) was highest for cows offered alfalfa silage, intermediate for clover silage, and lowest for cows offered grass silage. These differences reflect the higher rumen outflow rates for legume silages in comparison with grass silage. However, the effect of these differences on N-use efficiency (feed to milk) was probably quite small in comparison with effects of N intake. Although the biohydrogenation of alpha-linolenic acid was still high for red clover silage (86.1% compared with 94.3% for grass silage), there was a 240% increase in the proportion of alpha-linolenic acid passing through the rumen. This explains the increased recovery of alpha-linolenic acid from feed into milk with diets based on red clover silage.     

The effect of various doses of an exogenous acid protease on the fermentation and nutritive value of corn silage

The objective of this experiment was to evaluate the effects of treating whole-plant corn at harvest with various doses of an exogenous acidic protease on fermentation and changes in nutritive value after a short period (45 d) of ensiling. Whole-plant corn (37% dry matter) was chopped and treated with 0, 20, 200, 1,000, or 2,000 mg of protease/kg of wet forage. Forages (~500 g) were packed in bag silos and ensiled at 22 to 23°C for 45 d. Data were analyzed as a 5 × 2 factorial arrangement of treatments with the main effects of the dose of protease, day of ensiling, and their interaction. Treatment with protease did not alter the concentrations of dry matter, neutral detergent fiber, acid detergent fiber, starch, lactic acid, or acetic acid compared with untreated silage, with the exception that the concentration of starch was lower in silage treated with 20 mg of protease/kg compared with untreated silage. However, the 2 highest doses of protease resulted in silages with higher concentrations of ethanol and more yeasts compared with untreated silage. Protease treatment did not affect the ruminal in vitro digestibility of neutral detergent fiber. Concentrations of soluble protein (percentage of crude protein) increased after ensiling for all treatments but was not different between silage treated with the lowest dose of protease and untreated silage. Soluble protein increased in a dose-dependent manner above the low dose of protease in silages. Concentrations of NH₃-N were higher only in silages treated with the 2 highest doses of protease compared with untreated silage. Silages treated with the 3 highest doses of protease were higher in ruminal in vitro digestibility of starch compared with untreated silage but were similar to each other. The concentrations of total AA were determined in fresh forage and silages for the untreated and 200 and 2,000 mg/kg doses of protease. Neither amount of added protease affected the total concentrations of essential, nonessential, or total AA in silage. However, of the essential AA, treatment with protease resulted in silages with lower concentrations of lysine and arginine but higher concentrations of leucine compared with untreated silage. The 200 mg/kg dose of protease substantially improved ruminal in vitro starch digestion in corn silage after a short period of ensiling without affecting concentrations or numbers of ethanol and yeasts, respectively.     

Comparison of grass and legume silages for milk production. 1. Production responses with different levels of concentrate

Silages prepared from pure stands of ryegrass, alfalfa, white clover, and red clover over two successive year were offered to lactating dairy cows in two feeding experiments. Proportional mixtures of all cuts prepared in a yr were used to ensure that the forage treatments were representative of the crop. Additional treatments involved mixtures of grass silage with either white clover silage or red clover silage (50/50, on a DM basis). Silages were prepared in round bales, using a biological inoculant additive, and wilting for up to 48 h. Although the legumes were less suited to silage-making than grass, because of their higher buffering capacity and lower water-soluble carbohydrate content, all silages were well-fermented. A standard concentrate was offered at a flat-rate (8 kg/d in yr 1, and 4 or 8 kg/d in yr 2). All of the legume silages led to higher DM intake and milk yields than for the grass silage, with little effect on milk composition. Intake and production responses to legumes were similar at the two levels of concentrate feeding and with forage mixtures they were intermediate to those for the separate forages. An additional benefit of the clover silages, particularly red clover silage, was the increase in levels of polyunsaturated fatty acids, particularly alpha-linolenic acid, in milk. Legume silages also led to a lower palmitic acid percentage in milk. The efficiency of conversion of feed N into milk N declined with increasing levels of legume silage. White clover silage led to a higher N-use efficiency when the effect of N intake level is taken into account.     

Effects of silage species and supplemental vitamin E on the oxidative stability of milk

Two experiments were conducted to study the effects of feeding legume silages and providing supplemental vitamin E in concentrates on the oxidative stability of milk. In experiment 1, six multiparous Holstein-Friesian cows were offered 1 of 6 silage treatments in a cyclical changeover-design experiment, with four 4-wk periods. The silages were grass, red clover, white clover, alfalfa, grass and red clover mixture (50:50 on a DM basis), and grass and white clover mixture (50:50 on a DM basis). In experiment 2, 8 cows were used in a changeover-design experiment with three 4-wk periods. The 4 treatments were a factorial combination of forages (grass silage or red clover silage) and supplemental vitamin E in the form of all-rac-alpha-tocopheryl acetate (29 or 290 IU/kg of DM in the concentrate). All forages were offered ad libitum and a flat rate of concentrates (8 kg/d) was fed in both experiments. Red clover silage led to significantly higher forage intakes, milk yields, and milk protein percentage in experiment 2, which was in agreement with results from experiment 1. There was no effect of vitamin E on feed intake, milk production, or milk fat and protein percentage. Red clover silage also led to significant changes in milk fatty acid profiles, particularly increased levels of polyunsaturated fatty acids. Milk samples were stored at 4 degrees C and 20 degrees C and analyzed for alpha-tocopherol and thiobarbituric acid reactive substances at intervals to determine oxidative stability. Diets based on red clover and alfalfa silages were associated with more rapid loss of alpha-tocopherol and increased production of thiobarbituric acid reactive substances during the storage of milk in comparison with diets based on grass silage. The increased oxidative deterioration of milk produced from cows fed red clover silage was avoided by vitamin E supplementation.     

Degradation of Crude Protein in Forages Determined by In Vitro and In Situ Procedures

Objectives were to determine in selected forages fiber and protein fractions, in vitro and in situ CP degradability, and to compare in vitro methods of estimating rumen CP degradability with the in situ bag technique. Forages analyzed (five samples per treatment except alfalfa hays, which had four) included alfalfa as baled hay, alfalfa ensiled in conventional upright silos, alfalfa ensiled in oxygen-limiting silos, ammonia-treated corn silage, untreated corn silage, and orchardgrass hay. Untreated corn silages had the greatest protease insoluble CP at 48 h, indicating protein in corn silage is not degraded well by protease enzyme. In situ CP degradability was greater than 80% for all ensiled forages. Ensiled forages had the greatest estimated A fraction (rapidly degraded in the rumen), alfalfa hays had the greatest B fraction (intermediate rate of degradation), and orchardgrass hays had the greatest C fraction (not degraded in rumen).High correlations between in situ degradability and some in vitro measurements suggest laboratory techniques of estimating CP degradability of forages are possible. For silages, buffer-soluble CP had the greatest correlation (.58) with in situ degradability; for hays, NDIN had the greatest correlation (−.83).     

Effects of Nutrient Additives and Sodium Hydroxide on Ensiling Characteristics and In Vitro Fiber Digestion Kinetics of Dairy Cattle Waste-Energy Feedstuff Mixtures

Dairy waste fiber was mixed in ratio 1:1 (dry matter basis) with ground alfalfa hay, chopped cornstalks, chopped whole corn plant, or ground corn. Prior to ensiling, mixtures received either no additive, dry cane molasses at 5% of dry matter, urea at 1% of dry matter, dry cane molasses at 5% plus urea at 1% of dry matter, sodium hydroxide at 3.3% of dry matter, or sodium hydroxide at 3.3% plus urea at 1% of dry matter. Lactate concentration of silages increased progressively in those receiving alkali, urea, or molasses. Sodium hydroxide treatment tended to increase unavailable nitrogen of silages. Molasses increased a) digestion rate of acid detergent fiber in alfalfa: dairy waste fiber, and b) digestion coefficients of corn silage: and corn:dairy waste fiber silages. Urea additions resulted in less digestion of stalklage:dairy waste fiber. Addition of soluble nitrogen and energy together often improved digestion over addition of either one alone. Sodium hydroxide was effective in increasing a) digestion of certain fiber fractions of all silages except corn silage: dairy waste fiber and b) digestion rate of corn silage:dairy waste fiber hemicellulose. Addition of urea to alkali-treated corn silage:dairy waste fiber increased digestion of neutral detergent fiber and hemicellulose beyond that of alkali-treated mixtures alone.     

Fate of Escherichia coli O157:H7 and bacterial diversity in corn silage contaminated with the pathogen and treated with chemical or microbial additives

Inhibiting the growth of Escherichia coli O157:H7 (EC) in feeds may prevent the transmission or cycling of the pathogen on farms. The first objective of this study was to examine if addition of propionic acid or microbial inoculants would inhibit the growth of EC during ensiling, at silo opening, or after aerobic exposure. The second objective was to examine how additives affected the bacterial community composition in corn silage. Corn forage was harvested at approximately 35% dry matter, chopped to a theoretical length of cut of 10 mm, and ensiled after treatment with one of the following: (1) distilled water (control); (2) 1 × 10⁵ cfu/g of EC (ECCH); (3) EC and 1 × 10⁶ cfu/g of Lactobacillus plantarum (ECLP); (4) EC and 1 × 10⁶ cfu/g of Lactobacillus buchneri (ECLB); and (5) EC and 2.2 g/kg (fresh weight basis) of propionic acid, containing 99.5% of the acid (ECA). Each treatment was ensiled in quadruplicate in laboratory silos for 0, 3, 7, and 120 d and analyzed for EC, pH, and organic acids. Samples from d 0 and 120 were also analyzed for chemical composition. Furthermore, samples from d 120 were analyzed for ammonia N, yeasts and molds, lactic acid bacteria, bacterial community composition, and aerobic stability. The pH of silages from all treatments decreased below 4 within 3 d of ensiling. Escherichia coli O157:H7 counts were below the detection limit in all silages after 7 d of ensiling. Treatment with L. buchneri and propionic acid resulted in fewer yeasts and greater aerobic stability compared with control, ECCH, and ECLP silages. Compared with the control, the diversity analysis revealed a less diverse bacterial community in the ECLP silage and greater abundance of Lactobacillus in the ECLP and ECA silages. The ECLB silage also contained greater abundance of Acinetobacter and Weissella than other silages. Subsamples of silages were reinoculated with 5 × 10⁵ cfu/g of EC either immediately after silo opening or after 168 h of aerobic exposure, and EC were enumerated after 6 or 24 h, respectively. All silages reinoculated with EC immediately after silo opening (120 h) had similar low pH values (<4.0) and EC counts were below the detection limit. The ECCH and ECLP silages reinoculated with EC after 168 h of aerobic exposure had relatively high pH values (>5.0) and EC counts (5.39 and 5.30 log cfu/g, respectively) 24 h later. However, those treated with L. buchneri or propionic acid had lower pH values (4.24 or 3.96, respectively) and lower EC counts (1.32 log cfu/g or none, respectively). During ensiling, EC was eliminated from all silages at pH below 4.0. During aerobic exposure, the growth of EC was reduced or prevented in silages that had been treated with L. buchneri or propionic acid at ensiling, respectively.     

Red clover extracts inhibit legume proteolysis

Unwilted red clover and alfalfa herbage were hand-chopped, inoculated, ensiled into laboratory silos and opened after 0, 1, 3 and 7 days. Proteolysis was 40% lower with the red clover silage than alfalfa silage after 7 days of ensiling. The lower extent of proteolysis with the red clover silages could not be explained by differences in dry matter content, pH decline nor final pH of the silage. It was determined that red clover has a buffer soluble protein that could inhibit proteolysis in red clover and alfalfa. The soluble protein is possibly a polyphenol oxidase.