Aerobic deterioration of lucerne (Medicago sativa) and Maize (zea mais) Silages—Effects of Yeasts

Lucerne (Medicago sativa L) silages made in test tube silos at various dry matter (DM) levels (290-537 g kg^?1) with and without addition of glucose (20 g kg^?1 herbage) at ensiling were stable during 7 days of aerobic exposure. Lucerne silages taken from seven farm silos were similarly stable while three others were unstable (average DM loss of 62 g kg^?1 DM and pH increase from 4.35 to 7.58 in 7 days). Under similar circumstances, maize (Zea mais L) silage had a high DM loss (164 g kg^?1 DM) and an increase in pH from 3.94 to 8.13. Silages were inoculated with yeast (1 × 10⁶ CFU g^?1 silage) isolated from aerobically deteriorating farm-scale lucerne and maize silages. The stable lucerne silages were not destabilised by inoculation, but the instability of unstable lucerne silages was increased. Aerobically unstable maize silage and stable lucerne silage were inoculated and studied separately or as a 50:50 mixture. There was a distinct lag in the development of aerobic instability in the mixture compared with maize silage alone. In another experiment, the growth of the inoculum in malt agar (pH 3.5) as affected by the presence of fresh and ensiled lucerne (six cultivars), birdsfoot trefoil (Lotus corniculatus L) and red clover (Trifolium pratense L) was studied. The legume herbages did not inhibit yeast growth whereas the corresponding silages did. Five aerobically stable lucerne silages inhibited yeast development in the media whereas unstable maize silage did not. It is concluded that aerobic stability was not related to silage DM, pH, yeast numbers or glucose addition at ensiling. Stability appeared to be due to the presence of an inhibitor (or inhibitors) produced during ensilage.     

The use of pimaricin as an additive to improve the aerobic stability of silage

Pimaricin was added to samples from each of five crop species to give the following nominal rates of application: 0, 0.011, 0.022, 0.034, 0.045, 0.09 and 0.27 kg t^?1 fresh weight and the materials were ensiled in 100 g and 10 kg fresh weight quantities. Pimaricin applied at ensiling at 0.011 or 0.022 kg t^?1 reduced aerobic deterioration in three of the five crops—ryegrass, lucerne and tall fescue; a much higher rate of 0.27 kg t^?1 was required with silages made from red clover and maize. This beneficial effect occurred even though only from 1-66% of the pimaricin initially applied was recovered after the period of anaerobic storage in the silos. Pimaricin had no apparent effect on the course of fermentation nor on the composition of the silages, all of which were well preserved.     

Nutritive value of maize silage in relation to dairy cow performance and milk quality

Maize silage has become the major forage component in the ration of dairy cows over the last few decades. This review provides information on the mean content and variability in chemical composition, fatty acid (FA) profile and ensiling quality of maize silages, and discusses the major factors which cause these variations. In addition, the effect of the broad range in chemical composition of maize silages on the total tract digestibility of dietary nutrients, milk production and milk composition of dairy cows is quantified and discussed. Finally, the optimum inclusion level of maize silage in the ration of dairy cows for milk production and composition is reviewed. The data showed that the nutritive value of maize silages is highly variable and that most of this variation is caused by large differences in maturity at harvest. Maize silages ensiled at a very early stage (dry matter (DM) < 250 g kg^?1) were particularly low in starch content and starch/neutral detergent fibre (NDF) ratio, and resulted in a lower DM intake (DMI), milk yield and milk protein content. The DMI, milk yield and milk protein content increased with advancing maturity, reaching an optimum level for maize silages ensiled at DM contents of 300–350 g kg^?1, and then declined slightly at further maturity beyond 350 g kg^?1. The increases in milk (R² = 0.599) and protein (R² = 0.605) yields with maturity of maize silages were positively related to the increase in starch/NDF ratio of the maize silages. On average, the inclusion of maize silage in grass silage‐based diets improved the forage DMI by 2 kg d^?1, milk yield by 1.9 kg d^?1 and milk protein content by 1.2 g kg^?1. Further comparisons showed that, in terms of milk and milk constituent yields, the optimum grass/maize silage ratio depends on the quality of both the grass and maize silages. Replacement of grass silage with maize silage in the ration, as well as an increasing maturity of the maize silages, altered the milk FA profile of the dairy cows, notably, the concentration of the cis‐unsaturated FAs, C18:3n‐3 and n‐3/n‐6 ratio decreased in milk fat. Despite variation in nutritive value, maize silage is rich in metabolizable energy and supports higher DMI and milk yield. Harvesting maize silages at a DM content between 300 and 350 g kg^?1 and feeding in combination with grass silage results in a higher milk yield of dairy cows. © 2014 Society of Chemical Industry     

The effect of a cellulase preparation derived from Trichoderma viride on the chemical changes during the ensilage of grass,lucerne and clover

Perennial ryegrass, lucerne and clover were each treated with formic acid (4.5 litre t^?1 crop) and a commercial cellulase preparation derived from Trichoderma viride (4 g kg^?1) and ensiled in triplicate in test-tube silos, capacity 80 g. The crops were minced, finely chopped or coarsely chopped before ensiling and the test-tubes were held at 0, 15, 35 or 50°C. All the silages were well preserved and the grass silages, with and without cellulase treatment, contained residual water-soluble carbohydrates. Treatment with the cellulase preparation resulted in more cellulose being hydrolysed in the grass silages than in the legume silages. The greatest losses of cellulose in each crop occurred in the minced silages held at 15 and 35°C.     

Occurrence of mycotoxins in maize,grass and wheat silage for dairy cattle in the Netherlands

The occurrence of mycotoxins in 140 maize silages, 120 grass silages and 30 wheat silages produced in the Netherlands between 2002 and 2004 was determined using a liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) multi-method. Deoxynivalenol (DON) was detected above the limit of quantification (LOQ) of 250 μg kg^?1 in 72% of maize and 10% of wheat silages. Average DON concentrations were 854 and 621 μg kg^?1, respectively, and maximum concentrations 3142 and 1165 μg kg^?1, respectively. Zearalenone was detected above the LOQ of 25 μg kg^?1 in 49% of maize and 6% of grass silages. Average zearalenone concentrations were 174 and 93 μg kg^?1, respectively, and maximum concentrations 943 and 308 μg kg^?1, respectively. The incidences and average concentrations of DON and zearalenone in maize silage were highest in 2004. The incidence of other mycotoxins was low: fumonisin B1 and 15-acetyl-DON were detected in 1.4 and 5% of maize silages, respectively, and roquefortin C in 0.8% of grass silages. None of the silages contained aflatoxins, ochratoxin A, T2-toxin, HT2-toxin, sterigmatocystin, diacetoxyscirpenol, fusarenon-X, ergotamine, penicillinic acid, or mycophenolic acid. This study demonstrates that maize silage is an important source of DON and zearalenone in the diet of dairy cattle. Since the carryover of these mycotoxins into milk is negligible, their occurrence in feed is not considered to be of significant concern with respect to the safety of dairy products for consumers. Potential implications for animal health are discussed.     

The effects of the rate of addition of formic acid and sulphuric acid on the ensilage of perennial ryegrass in laboratory silos

Perennial ryegrass was ensiled in laboratory silos after addition of formic acid (850 g kg^?1) or sulphuric acid (906 g kg^?1) at rates of 0, 2, 4 and 6 litres t^?1 fresh grass. Silos were opened after 6, 18 and 90 days and the silage subjected to chemical and microbiological analysis. The untreated control was poorly fermented with a final pH of 4.7, a butyric acid concentration of 19 g kg^?1 dry matter (DM) and an NH₃-N content of 275 g kg^?1 total nitrogen (TN). For the formic acid treatments the 2 litre t^?1 and 6 litre t^?1 levels both produced well-preserved silages but they were of different types. The silage treated with 2 litre t^?1 had a pH of 4.0, a lactic acid concentration of 92 g kg^?1 DM and 161 g NH₃-N kg^?1 TN, whereas with the 6 litre t^?1 treatment, fermentation had been severely restricted. The pH was 4.2, the lactic acid concentration was only 8 g kg^?1 DM and the NH₃-N content was 80 g kg^?1 TN. However, formic acid at 4 litre t^?1 produced a badly fermented silage of final pH 5.0 with lactic acid and butyric acid concentrations of 16 and 15 g kg^?1 DM, respectively, and an NH₃-N content of 149 g kg^?1 TN. Sulphuric acid at 2 and 4 litres t^?1 produced silages of low lactic acid contents, 36 and 24 g kg^?1 DM, and they also contained butyric acid in concentrations of 13 and 11 g kg^?1 DM; respective NH₃-N contents were 206 and 114 g kg^?1 DM. When sulphuric acid was added at 6 litres t^?1, despite a reduction in the pH of the grass to 3.5, fermentation was not restricted as it was with the equivalent level of formic acid. Lactic acid was present at 27 g kg^?1 DM and the ethanol concentration was very high at 66 g kg^?1 DM; the sulphuric acid-treated silages were characterised by high yeast counts. At the higher rates of addition, formic acid reduced the.     

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     

Changes during ensilage in the nitrogenous components of fresh and additive treated ryegrass and lucerne

The changes in the major nitrogenous components of herbage during the ensilage of lucerne and Italian ryegrass, with and without the addition of 85% formic acid and 40% formaldehyde at the rates of 3.2 and 7.4 g kg^?1 fresh herbage respectively, were studied. Both herbages were of similar amino-N composition except for asparagine which was about 10% of total nitrogen (TN) in lucerne and only 0.2% of TN in Italian ryegrass. After 90 days ensiling about 90% of the sum of protein-, peptide-, amino- and amide-N was preserved in the additive-treated silages while only 58% was recovered in the untreated lucerne (butyrate) silage. Recoveries of these compounds in the untreated ryegrass (lactate) silage was 83%. In the butyrate silage, the branched chain amino acids, valine, leucine and isoleucine were well preserved. In addition to these, proline, phenylalanine, glycine, methionine and threonine were well preserved in the lactate silages. Both the untreated silages contained more alanine than in their original respective herbages. Most of the losses from the amine-forming amino acids were recovered as their corresponding amines. Alpha-amino butyric acid and δ-amino valeric acid were found in both the untreated silages and the quantity present was dependent on the silage quality. In both the additive-treated silages, recoveries of amino acids, other than sulphur amino acids, lysine, histidine, tyrosine and tryptophan, were ca 90% or greater. The losses of these amino acids, excluding the sulphur amino acids, can be attributed mainly to the direct effects of formaldehyde in preventing their estimation following freeze drying or acid hydrolysis.     

Effects of peptidase inhibitors and other additives on fermentation and nitrogen distribution in perennial ryegrass silage

The effects of formic acid, three concentrations of formaldehyde in a formic acid/formaldehyde mixture and cysteine‐peptidase inhibitors, 1‐trans epoxysuccinyl‐leucylamido‐(4‐guanidino) butane (E‐64), N‐ethylmaleimide and cystamine on nitrogen (N) distribution during ensilage of perennial ryegrass (Lolium perenne) were investigated. A third cut or perennial ryegrass (163 g dry matter kg⁻¹ and 61 g water‐soluble carbohydrate kg⁻¹ dry matter) was ensiled in two silo sizes; formaldehyde‐treated herbage was ensiled only in larger silos (500–550 g herbage) and cysteine‐peptidase inhibitor‐treated herbage only in smaller silos (130–150 g herbage). Control silages were poorly fermented but contained low concentrations of butyric acid and ammonia N indicating little activity of spoilage bacteria. Formic acid increased peptide N concentrations (P<0.01) in silage from smaller silos but had little effect on other N constituents; in the larger silos, formic acid reduced soluble non protein nitrogen (NPN) and ammonia‐N concentrations and increased peptide N concentrations. Increments in formaldehyde reduced silage soluble and ammonia N concentrations (linear effect; P<0.001). N‐Ethylmaleimide and E‐64 reduced soluble NPN concentrations (P<0.05) but had little effect on other N constituents. Cystamine, however, increased silage peptide N concentrations. Gel filtration on Sephadex G‐25 of silage juice prepared from control and formic acid‐treated silages suggested that most silage peptides were small, with molecular weights of less than 520 Da. © 1999 Society of Chemical Industry     

The potential of sunflower as a crop for ensilage

The chemical composition and nutritive value of sunflower, cv. Armavirec, was examined at 12 stages of growth. Maximum yield of dry matter (18.2 t ha^?1) occurred at the dough seed stage. Ether extract showed a five-fold increase with the development of the seed. This increase was reflected in the gross energy value which was maximal at 19.1 MJ kg^?1 dry matter at the dough seed stage. In-vitro organic matter digestibility was highest at the onset of flowering when the estimated metabolisable energy value was 10.5 MJ kg^?1 dry matter. Throughout the sampling period the calcium to phosphorus ratios varied from 3.9-5.9:1, an imbalance which would have to be redressed on diets based largely on sunflowers. Fresh and wilted sunflowers were successfully ensiled at the flowering stage, without the use of additives. pH values of the resulting silages were 3.86 and 4.01. The silages were each fed to six cross-bred wethers and voluntary dry matter intakes were 23.9 and 26.5 g kgW^?1 for the fresh and wilted silages respectively. For the fresh silage, intakes of digestible organic matter and digestible nitrogen were 612 and 13.6 g day^?1 compared with 684 and 16.7 g day^?1 for the wilted silage. Nitrogen retention increased with increase in silage dry matter. The optimum time of cutting for yield of nutrients is at the milky ripe stage. The composition of the crop at this stage should facilitate a satisfactory preservation by ensiling. The crop provides a high yield of dry matter over about 10 weeks in late summer and could provide a valuable standby forage crop for grass in dry areas.     

The effect of decanoic,dodecanoic and tetradecanoic acids on silage fermentation

The ability of decanoic (C₁₀), dodecanoic (C₁₂) and tetradecanoic (C₁₄) acids to restrict the fermentation of ryegrass and lucerne was investigated with laboratory silos. With ryegrass, addition of the C₁₂ acid alone at 1 kg t^?1 of fresh crop had no effect on extent or pattern of fermentation compared to that of the untreated crop, though at 2 kg t^?1 the acid appeared to reduce the content of butyric acid but not ammonia-N in the silage. Formic acid, added alone or with C₁₀, C₁₂ and C₁₄ acids restricted extent of fermentation and butyric acid production, but in this experiment addition of these acids had relatively little effect on the content of ammonia-N in the silages. With lucerne, extent of fermentation was similar for all treatments, though the untreated silage and those which received C₁₂ or formic acid alone at 1 kg t^?1 or 1 kg t^?1 of C₁₄ acid with 1 kg t^?1 of formic acid contained butyric acid in excess of 3% of the dry matter (DM). It is concluded that acids in the range C₁₂-C₁₄ are not effective in restricting silage fermentation and that their effectiveness is not increased by reduction in pH produced by the addition of formic acid. Microbiological assessments largely confirmed these findings.     

Effect of the stage of growth,wilting and inoculation in field pea (Pisum sativum L.) silages. II. Nitrogen fractions and amino acid compositions of herbage and silage

The aim of the study was to investigate the effects of stage of maturity, wilting and inoculant application on proteolysis in field pea herbage and silage in northern Italy. Semi‐leafless field peas were harvested at four progressive morphological stages (end of flowering, I; beginning of pod filling, II; advanced pod filling, III; beginning of ripening, IV). For each stage of growth, the herbage was field wilted to a dry matter (DM) content around 330 g kg^?1. The unwilted and wilted herbages were ensiled with (I) and without (C) a lactic acid bacteria (LAB) inoculant. The stage of growth significantly affected all the nitrogen fraction concentrations both in herbage and in silage. Extensive proteolysis took place in silages made at early morphological stages and characterised by a high crude protein content (around 240 g kg^?1 DM), as shown by the non‐protein nitrogen (NPN) value (around 700 g kg^?1 total N or higher). Neither inoculum nor wilting helped to reduce the protein degradation in these silages. The level of proteolysis was reduced in silages made from the pod filling stage onwards where most of the protein was localised in the seed and a better fermentation pattern was observed. The amino acid (AA) composition of silages made at stage IV was close to that of fresh herbage, with minimal losses of nutritionally essential amino acids for ruminants. Copyright © 2006 Society of Chemical Industry     

The occurrence and prevention of ethanol fermentation in high‐dry‐matter grass silage

Ethanol is a common, usually minor fermentation product in ensiled forages, the major product being lactic acid. Occasionally, high levels of ethanol are found in silages. The aim of this study was to determine the incidence of high‐dry‐matter (DM) grass silages containing ethanol as the main fermentation product (ethanol silages), to describe the fermentation process in such silages and to determine the effect of grass maceration prior to wilting and addition of a bacterial inoculant containing Lactobacillus plantarum and Enterococcus faecium strains on fermentation. Twenty‐one laboratory silages produced between 1993 and 1995, 21 farm silages produced between 1980 and 1989 and 36 farm silages produced in 1995 (all produced without additive) were examined for pH and chemical composition. Dry matter (DM) loss during ensilage was determined for the laboratory silages only. Four laboratory silages were identified as ethanol silages. Mean concentrations of ethanol, lactic acid and acetic acid were 48.1, 15.5 and 6.0 g kg⁻¹ DM respectively. In the silages that contained lactic acid as the main fermentation product (lactic acid silages) these values were 7.7, 45.5 and 15.1 g kg⁻¹ DM. Mean DM loss and pH were 62.8 g kg⁻¹ DM and 5.32 respectively for ethanol silages and 24.4 g kg⁻¹ DM and 4.69 for lactic acid silages. There was no difference between ethanol silages and lactic acid silages in the mean concentration of ammonia‐N (94 g kg⁻¹ total N), and butyric acid was not detected (<0.2 g kg⁻¹ DM), indicating that both types of silages were well preserved. Analysis of the composition of the grass at ensiling showed a positive correlation between the concentration of soluble carbohydrates and the development into ethanol silage. Analysis of the farm silages indicated that 29% of the silages produced between 1980 and 1989 and 14% of those produced in 1995 were ethanol silages. Maceration prior to wilting and addition of silage inoculant improved lactic acid fermentation and prevented high ethanol levels. The micro‐organisms responsible for ethanol fermentation as well as the implications of feeding ethanol silages to livestock remain to be resolved. © 2000 Society of Chemical Industry     

The influence of hydrochloric acid on the effect of caproic acid in preventing aerobic deterioration of silages

In five experiments of the same design using silages made from wilted Italian ryegrass (dry matter 22.7–40.9%), the effects of caproic acid (C₆) (10 mmol kg^?1) with and without 6_N-HCl (20 ml kg^?1), applied at ensiling or at opening, in preventing aerobic deterioration of silages after opening silos were investigated. Application of C₆ either at ensiling or at opening was partially effective in preventing temperature increase, pH rise and breakdown of organic acids during the 7-day aerobic exposure period after opening the silos. Addition of HCl reduced the pH of silages irrespective of the time of application. However, no improvement of the effect of C₆ due to HCl supplementation was observed as far as the prevention of the aerobic deterioration was concerned. Treatment with C₆+ HCl at ensiling inhibited silage fermentation more extensively than C₆ alone. Viable counts of yeasts and moulds in the silages treated with C₆ or C₆+ HCl at ensiling tended to be lower than those in the control silages. Considerable increases in the counts of moulds were found in the control silages during the aerobic exposure period.     

Aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure

BACKGROUND: Most of the maize silage stored in horizontal silos is exposed to air and can be spoiled by fungi. Potentially toxigenic fungi have been found in maize silage, and about 300 mycotoxins have been detected. Among these mycotoxins, the most harmful for feed and food safety are aflatoxins. The aim of the study was to set up a specific method to detect aflatoxins in maize silage, and to investigate whether aflatoxin contamination in maize silage depends on the level of field contamination of the crop, and whether the occurrence of aerobic spoilage during ensiling has any effect on the final contamination of the silage. RESULTS: A method for the determination of aflatoxin B₁, B₂, G₁ and G₂ in maize silage using high‐performance liquid chromagraphy with fluorescence detection has been developed and validated. Recoveries of aflatoxin B₁, B₂, G₁, and G₂ spiked over the 0.25 to 5 µg kg^?1 range averaged 74–94%. The results of laboratory scale and farm scale ensiling experiments indicated that aflatoxins could increase when silage is exposed to air during conservation or during the feed‐out phase. CONCLUSIONS: The method here proposed to detect aflatoxins in silages has proved to be sensitive and is able to detect levels of 0.1 and 0.5 ng mL^?1 for AFB₁ and AFG₁, and between 0.025 and 0.125 ng mL^?1 for AFB₂ and AFG₂. This study also provides evidence of aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure. Copyright © 2011 Society of Chemical Industry     

Production of lactating dairy cows fed alfalfa or red clover silage at equal dry matter or crude protein contents in the diet

Two Latin square trials, using 21 or 24 multiparous lactating Holstein cows, compared the feeding value of red clover and alfalfa silages harvested over 2 yr. Red clover silages averaged 2 percentage units lower in crude protein (CP) and more than 2 percentage units lower in neutral detergent fiber and acid detergent fiber than did alfalfa silage. In trial 1, diets were formulated to 60% dry matter (DM) from alfalfa, red clover silage, or alfalfa plus red clover silage (grown together); CP was adjusted to about 16.5% by adding soybean meal, and the balance of dietary DM was from ground high moisture ear corn. Nonprotein N in red clover and alfalfa-red clover silages was 80% of that in alfalfa silage. Although DM intake was 2.5 and 1.3 kg/d lower on red clover and alfalfa plus red clover, yield of milk and milk components was not different among diets. In trial 2, four diets containing rolled high moisture shelled corn were formulated to 60% DM from alfalfa or red clover silage, or 48% DM from alfalfa or red clover silage plus 12% DM from corn silage. The first three diets contained 2.9% soybean meal, and the red clover-corn silage diet contained 5.6% soybean meal; the 60% alfalfa diet contained 18.4% CP, and the other three diets averaged 16.5% CP. Nonprotein N in red clover silage was 62% of that in alfalfa silage. Intake of DM was about 2 (no corn silage) and 1 kg/d (plus corn silage) lower on red clover. Yield of milk and milk components was not different among the first three diets; however, yields of milk, total protein, and true protein were higher on red clover-corn silage with added soybean meal. Replacing alfalfa with red clover improved feed and N efficiency and apparent digestibility of DM, organic matter, neutral detergent fiber, acid detergent fiber, and hemicellulose in both trials. Net energy of lactation computed from animal performance data was 18% greater in red clover than alfalfa. Data on milk and blood urea and N efficiency suggested better N utilization on red clover.     

Reduction of proteolysis during ensilage of perennial ryegrass by protease inhibitors

Two experiments were carried out to assess the effectiveness of protease inhibitors in reducing proteolysis during ensilage. Perennial ryegrass (Lolium perenne) (279 g dry matter (DM) kg^?1, experiment 1 and 170 gDM kg^?1 experiment 2) was ensiled in laboratory silos for 82 and 50 days, respectively. With the exception of the application of formic acid (1.5 g kg^?1) plus formalin (3 g kg^?1) in experiment 1, all silages had pH values of less than 4.0 and lactic acid concentrations of greater than 100 g kg^?1 DM. Application of the cysteine endopeptidase inhibitor E-64 (1-trans-epoxysuccinyl-leucylamido(4-guanidino) butane, 22.5 mg kg^?1 reduced proteolysis during ensilage as measured by a reduction in the quantities of soluble non-protein N in silage to 0.76 and 0.91 of control values (528 and 643 g kg^?1 total N) in experiments 1 and 2, respectively. In experiment 2 increasing the application rate of E-64 from 22.5 to 45 mg kg^?1 further reduced the extent of proteolysis from 0.91 to 0.75 of control soluble non-protein N concentrations. Proteolysis was also reduced in experiment 2 by two other cysteine endopeptidase inhibitors, antipain (32 mg kg^?1, 0.81 of control) and cystain (1.2 mg kg^?1; 0.89 of control). Pepstatin A (300 mg kg^?1), an inhibitor of aspartic acid endopeptidases, did not reduce soluble non-protein N concentrations in experiment 1 but did do so in experiment 2 (0.81 of control). Treatment of grass at ensiling with Lactobacillus plantarum (10⁶ g^?1) reduced silage non-protein N concentrations to 0.83 of the control. The effects of E-64 and pepstatin A and of E-64 and L plantarum in experiment 2 were addive. Cysteine endopeptidases therefore play an important role in protein breakdown during ensilage.     

Methane emission,nutrient digestibility,and rumen microbiota in Holstein heifers fed 14 different grass or clover silages as the sole feed

This experiment investigated the variation in enteric methane production and associated gas exchange parameters, nutrient digestibility, rumen fermentation, and rumen microbiome when a range of silages based on different forage types (grass or clover), and different species within the 2 types, were fed as the sole feed to heifers. Three grass species (perennial ryegrass, festulolium, and tall fescue) and 2 clover species (red clover and white clover) were included. Perennial ryegrass was harvested at 2 maturity stages in the primary growth, white clover was harvested once in the primary growth, and 4 cuts of festulolium and tall fescue and 3 cuts of red clover were harvested during the growing season, giving 14 different silage batches in total. Sixteen Holstein heifers 16 to 21 mo old and 2 to 5 mo in pregnancy were fed the silages ad libitum as the sole feed in an incomplete crossover design. Each silage was fed to 4 heifers, except for the 2 perennial ryegrass silages, which were fed to 8 heifers; in total 64 observations. The CH₄ production was measured for 3 d in respiration chambers. Heifers fed clover silages had higher dry matter intake (DMI) compared with heifers fed grass silages, and heifers fed tall fescue silages had the numerically the lowest DMI. Compared with grass silages, feeding clover silages led to higher crude protein digestibility but lower neutral detergent fiber (NDF) digestibility. Rumen pH was higher in heifers fed clover silages compared with those fed grass silages. Based on composition analysis, the rumen microbiota of the heifers clustered clearly according to forage type and species. More specifically, 7 of the 34 dominating rumen bacterial genus-level groups showed higher relative abundances for the clover silages, whereas 7 genus-level groups showed higher abundances for the grass silages. Methane yield was higher for heifers fed grass silages than for those fed clover silages when methane production was related to dry matter and digestible organic matter intake, whereas the opposite was seen when related to NDF digestion. The gross energy lost as methane (CH₄ conversion factor, %) reduced from 7.5% to 6.7%, equivalent to an 11% reduction. The present study gives the outlines for choosing the optimal forage type and forage species with respect to nutrient digestibility and enteric methane emission in ruminants.     

Characterisation of peptides in silages made from perennial ryegrass with different silage additives

The effects of applying either formic acid (5.4 g kg⁻¹), a mixture of formic acid (2.7 g kg⁻¹) and formaldehyde (1.5 g kg⁻¹, 81 g kg⁻¹ herbage crude protein) or two concentrations of a cysteine peptidase inhibitor, cystamine (5 or 50 g kg⁻¹), to perennial ryegrass (Lolium perenne) on the nitrogen (N) distribution of the resulting silages were investigated, with emphasis on changes in concentration, composition and molecular weight of silage peptides. Herbage (156 g dry matter kg⁻¹ and 141 g water‐soluble carbohydrate kg⁻¹ dry matter) was ensiled in triplicate in laboratory silos for 100 days. Formic acid and the formic acid/formaldehyde mixture reduced soluble non‐protein N and ammonia N concentrations (P < 0.01); in addition, formic acid increased peptide N concentrations (P < 0.05). Cystamine at 50 g kg⁻¹ reduced ammonia N concentrations (P < 0.01) and increased peptide N concentrations (P < 0.05), but when applied at 5 g kg⁻¹ had little effect. Gel filtration of silage extracts on Sephadex G‐25 suggested that a small proportion (0.06–0.11 g kg⁻¹ peptide N) of silage peptides had a chain length of 7–9 amino acids, but remaining peptides were smaller with chain lengths of 2–6 amino acid residues. Amino acid analysis of silage peptides indicated that additive treatment had little effect on peptide amino acid composition but that peptides with a chain length of 7–9 amino acids contained lower proportions of isoleucine and arginine. © 2000 Society of Chemical Industry     

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.