Sodium hydroxide enhances extractability and analysis of proanthocyanidins in ensiled sainfoin (onobrychis viciifolia)

Little information exists on the effects of ensiling on condensed tannins or proanthocyanidins. The acetone–butanol–HCl assay is suitable for measuring proanthocyanidin contents in a wide range of samples, silages included, but provides limited information on proanthocyanidin composition, which is of interest for deciphering the relationships between tannins and their bioactivities in terms of animal nutrition or health. Degradation with benzyl mercaptan (thiolysis) provides information on proanthocyanidin composition, but proanthocyanidins in several sainfoin silages have proved resistant to thiolysis. We now report that a pretreatment step with sodium hydroxide prior to thiolysis was needed to enable their analysis. This alkaline treatment increased their extractability from ensiled sainfoin and facilitated especially the release of larger proanthocyanidins. Ensiling reduced assayable proanthocyanidins by 29%, but the composition of the remaining proanthocyanidins in silage resembled that of the fresh plants.

Apparent digestibility and nitrogen utilization of diets based on maize silage harvested at three stages of maturity and fed to steers

Advancing maize crop maturity is associated with changes in ear-to-stover ratio which may have consequences for the digestibility of the ensiled crop. The apparent digestibility and nitrogen retention of three diets (Early, Mid and Late) containing maize silages made from maize of advancing harvest date [dry matter (DM) contents of the maize silages were 273, 314 and 367 g kg(-1) for the silages in the Early, Mid and Late diets respectively], together with a protein supplement offered in sufficient quantities to make the diets isonitrogenous, were measured in six Holstein-Friesian steers in an incomplete Latin square design with four periods. Dry-matter intake of maize silage tended to be least for the Early diet and greatest for the Medium diet (P=0(.)182). Apparent digestibility of DM and organic matter did not differ between diets. Apparent digestibility of energy was lowest in the Late diet (P = 0(.)057) and the metabolizable energy concentrations of the three silages were calculated as 11(.)0, 11(.)1 and 10(.)6 MJ kg(-1) DM for the Early, Medium and Late diets respectively (P = 0(.)068). No differences were detected between diets in starch digestibility but the number of undamaged grains present in the faeces of animals fed the Late diet was significantly higher than with the Early and Mid diets (P = 0(.)006). The apparent digestibility of neutral-detergent fibre of the diets reduced significantly as silage DM content increased (P = 0(.)012) with a similar trend for the apparent digestibility of acid-detergent fibre (P = 0(.)078). Apparent digestibility of nitrogen (N) was similar for the Early and Mid diets, both being greater than the Late diet (P = 0(.)035). Nitrogen retention did not differ between diets. It was concluded that delaying harvest until the DM content is above 300 g kg(-1) can negatively affect the nutritive value of maize silage in the UK.

Intake, live-weight gain and carcass characteristics of beef cattle given diets based on forage maize silage harvested at different stages of maturity

Advancing maturity of forage maize is associated with increases in the proportion of dry matter (DM) and starch, and decreases in the proportions of structural carbohydrates in the ensiled crop. This experiment investigated the effects of three maize silages of 291 (low), 339 (medium) and 393 (high) g DM per kg fresh weight on the performance of 48 Simmental. Holstein-Friesian cattle. Equal numbers of steers (mean start weight = 503 (s.d. 31.3) kg) and heifers (mean start weight = 378 (s.d. 11.2) kg) were offered individually isonitrogenous diets composed of the three silages plus a protein supplement with minerals once daily until slaughter at the target live weight of 575 and 475 kg for steers and heifers, respectively. Intake was reduced on the low diet (P < 0.01) compared with the other two treatments. Dietary starch intake increased by a total of 1 kg/day between low and medium diets but by only 0.2 kg/day between medium and high diets. Unlike starch intake, total neutral-detergent fibre intake showed no significant difference (P > 0.05) between diets. There were no differences in live-weight gain between treatments but differences (P < 0.05) in food conversion efficiency indicated relative gains of 115, 100 and 102 g gain per kg DM intake for diets low, medium and high, respectively. There were no differences between diets in carcass weights, fat score and overall conformation.

In vitro evaluation of fibrolytic enzymes as additives for maize (Zea mays L.) silage - II. Effects on rate of acidification, fibre degradation during ensiling and rumen fermentation

A study was carried out to determine the influence of fibrolytic enzymes derived from mesophilic or thermophilic fungal sources, added at ensiling, on time-course fermentation characteristics and in vitro rumen degradation of maize silage. The mesophilic enzyme was a commercial product derived from Trichodenna reesei (L), whereas the thermophilic enzyme was a crude extract produced from Thermoascus aurantiacus (Ta) in this laboratory. The fungus was cultured using maize cobs as a carbon source. The resulting fermentation extract was deionised to remove sugars and characterised for its protein concentration, main and side enzymic activities, optimal pH, protein molecular mass and isoelectric point. In an additional study, both enzymes were added to maize forage (333.5 g DM/kg, 70.0, 469.8, 227.1 and 307.5 g/kg DM of CP, NDF, ADF and starch, respectively) at two levels each, normalized according to xylanase activity, and ensiled in 0.5 kg capacity laboratory minisilos. Duplicate silos were opened at 2, 4, 8, 15, and 60 days after ensiling, and analysed for chemical characteristics. Silages from 60 days were bulked and in vitro gas production (GP) and organic matter degradability (OMD) profiles evaluated using the Reading Pressure Technique (RPT), in a completely randomised design. The crude enzyme extract contained mainly xylanase and endoglucanase activities, with very low levels of exoglucanase, which probably limited hydrolysis of filter paper. The extract contained three major protein bands of between 29 and 55 kDa, with mainly acidic isoelectric points. Ensiling maize with enzymes lowered (P < 0.05) the final silage pH, with this effect being observed throughout the ensiling process. All enzyme treatments reduced (P < 0.05) ADF contents. Treatments including Ta produced more gas (P < 0.05) than the controls after 24 h incubation in vitro, whereas end point gas production at 96 h was not affected. Addition of Ta increased (P < 0.01) OMD after 12 h (410 and 416 g/kg versus 373 g/kg), whereas both L and Ta increased (P < 0.05) OMD after 24 h. Addition of enzymes from mesophilic or thermophilic sources to maize forage at ensiling increased the rate of acidification of the silages and improved in vitro degradation kinetics, suggesting an improvement in the nutritive quality. (C) 2003 Elsevier B.V All rights reserved.

In vitro evaluation of fibrolytic enzymes as additives for maize (Zea mays L.) silage - III. Comparison of enzymes derived from psychrophilic, mesophilic or thermophilic sources

A completely randomised study was completed to examine the influence of fibrolytic enzymes derived from psychrophilic, (F), mesophilic, (L) or thermophilic (Ta) sources, applied at ensiling, on the chemical characteristics and in vitro rumen fermentation of maize silage, assessed using the Reading Pressure Technique (RPT). Treatments, all in triplicate, consisted of untreated maize forage or treated with preparations F, L, Ta or a mixture (1: 1, v/v) of F and L (FL), at two levels each, and ensiled for 210 days in plastic mini-silos. Addition of enzymes L decreased (P < 0.05) silage pH relative to the control, whereas enzyme Ta tended (P < 0.10) to reduce it. Preparations F, L and Ta tended to reduce (P < 0.10) the fibre contents of the silages, with effects being attributable to a decrease in the cellulose fraction. Starch contents were reduced (P < 0.05) in the treatments including enzyme F. End-point (96 h) gas production (GP) values did not differ among treatments, suggesting that enzymes did not change the total amount of fermentable substrate. However, consistent with the decrease in starch contents, adding enzyme F reduced (P < 0.05) GP at most incubation times. Addition of enzymes increased (P < 0.05) the initial (6 h) organic matter degradation (OMD) levels in all but one treatment (F), with increases of 14, 19, and 26% for preparations L, Ta, and FL, respectively, averaged across levels. Furthermore, the addition of enzymes increased (P < 0.05) the soluble OM losses, however, these increases did not fully account for the initial increase in OMD. The latter suggests that enzymes increased solubility and also altered silage structure, making it more amenable to degradation by ruminal microorganisms. As a result of the increase in OMD, without a concomitant increase in GP, the fermentation efficiency was greatly increased (P < 0.05) in enzyme treatments. Addition of enzymes to maize at ensiling, particularly those from the mesophilic and thermophilic sources used here, have the potential to increase the initial rate of silage OMD. (C) 2003 Elsevier B.V. All rights reserved.

Digestion, rumen fermentation and circulating concentrations of insulin, growth hormone and IGF-1 in steers given maize silages harvested at three stages of maturity

Advancing maturity of forage maize is associated with increases in the proportion of dry matter (DM) and starch and decreases in the proportions of structural carbohydrates in the ensiled crop. Three maize silages (286 (low, L), 329 (medium, M) and 379 (high, H) g DM per kg fresh weight) plus a concentrate formulated to give isonitrogenous intakes were offered to Holstein-Friesian steers fitted with a cannula in the dorsal sac of the rumen and a 'T' piece cannula in the proximal duodenum in an experiment with a cross-over design that allowed four collection periods. Nutrient flow to the duodenum was estimated using chromium-EDTA. Steers consumed approximately 0(.)6 kg DM per day less of diet L compared with the other two diets (P=0(.)026), resulting in less DM being digested (P=0(.)005) but digestibility did not differ between diets. Similar results were obtained for organic matter. There were no differences between diets in the intake or digestibility of neutral-detergent fibre. Intake, duodenal flow and faecal output of starch were greater for steers offered diets M and H compared with those given diet L (P < 0(.)05). In all diets rumen digestion contributed to over 90% of total digestion of starch, although rumen digestibility declined significantly with advancing maize maturity (P=0(.)002). Molar proportions of acetic acid were higher in diet H (P < 0(.)05) whilst proportions of propionic acid and n-butyric acid were higher in diets M and L. There were no significant differences between diets in mean rumen pH or ammonia concentrations. Mean circulating concentrations of insulin were higher (P=0(.)009) in cattle given diets L and M compared with diet H. There were no differences between diets in the mean circulating concentration of growth hormone, or the frequency, amplitude and duration of growth hormone pulses, or the mean circulating concentrations of IGF-1. Changes in forage composition that accompany advancing maize maturity affect overall silage digestion and circulating concentrations of insulin.

Alternative methodologies - stretching the in vitro box

Current gas-based in vitro evaluation systems are extremely powerful research techniques. However they have the potential to generate a great deal more than simple fermentation dynamics. Details from four experiments are presented in which adaptation, and novel application, of an in vitro system allowed widely differing objectives to be examined. In the first two studies, complement methodologies were utilised. In such assays, an activity or outcome is inferred through the occurrence of a secondary event rather than by direct observation. Using an N-deficient incubation medium, the increase in starch fermentation, when supplemented with individual amino acids (i.e., known level of N) relative to that of urea (i.e., known quantity and N availability), provided an estimate of their microbial utilisation. Due to the low level of response observed with some arnino acids (notably methionine and lysine), it was concluded, that they may not need to be offered in a rumen-inert form to escape rumen microbial degradation. In another experiment, the extent to which degradation of plant cell wall components was inhibited by lipid supplementation was evaluated using fermentation gas release profiles of washed hay. The different responses due to lipid source and level of inclusion suggested that the degree of rumen protection required to ameliorate this depression was supplement dependent. That in vitro inocula differ in their microbial composition is of little interest per se, as long as the outcome is the same (i.e., that similar substrates are degraded at comparable rates and end-product release is equivalent). However where a microbial population is deficient in a particular activity, increasing the level of inoculation will have no benefit. Estimates of hydrolytic activity were obtained by examining fermentation kinetics of specific substrates. A number of studies identified a fundamental difference between rumen fluid and faecal inocula, with the latter having a lower fibrolytic activity, which could not be completely attributed to microbial numbers. The majority of forage maize is offered as an ensiled feed, however most of the information on which decisions such as choice of variety, crop management and harvesting date are made is based on fresh crop measurements. As such, an attempt was made to estimate ensiled maize quality from an in vitro analysis of the fresh crop. Fermentation profiles and chemical analysis confirmed changes in crop composition over the growing season, and loss of labile carbohydrates during ensiling. In addition, examination of degradation residues allowed metabolizable energy (ME) contents to be estimated. Due to difficulties associated with starch analysis, the observation that this parameter could be predicted by difference (together with an assumed degradability), allowed an estimate of ensiled maize ME to be developed from fresh material. In addition, the contribution of the main carbohydrates towards ME showed the importance of delaying harvest until maximum starch content has been achieved. (c) 2005 Elsevier B.V. All rights reserved.

Simulating the effects of grassland management and grass ensiling on methane emission from lactating cows

A dynamic, mechanistic model of enteric fermentation was used to investigate the effect of type and quality of grass forage, dry matter intake (DMI) and proportion of concentrates in dietary dry matter (DM) on variation in methane (CH(4)) emission from enteric fermentation in dairy cows. The model represents substrate degradation and microbial fermentation processes in rumen and hindgut and, in particular, the effects of type of substrate fermented and of pH oil the production of individual volatile fatty acids and CH, as end-products of fermentation. Effects of type and quality of fresh and ensiled grass were evaluated by distinguishing two N fertilization rates of grassland and two stages of grass maturity. Simulation results indicated a strong impact of the amount and type of grass consumed oil CH(4) emission, with a maximum difference (across all forage types and all levels of DM 1) of 49 and 77% in g CH(4)/kg fat and protein corrected milk (FCM) for diets with a proportion of concentrates in dietary DM of 0.1 and 0.4, respectively (values ranging from 10.2 to 19.5 g CH(4)/kg FCM). The lowest emission was established for early Cut, high fertilized grass silage (GS) and high fertilized grass herbage (GH). The highest emission was found for late cut, low-fertilized GS. The N fertilization rate had the largest impact, followed by stage of grass maturity at harvesting and by the distinction between GH and GS. Emission expressed in g CH(4)/kg FCM declined oil average 14% with an increase of DMI from 14 to 18 kg/day for grass forage diets with a proportion of concentrates of 0.1, and on average 29% with an increase of DMI from 14 to 23 kg/day for diets with a proportion of concentrates of 0.4. Simulation results indicated that a high proportion of concentrates in dietary DM may lead to a further reduction of CH, emission per kg FCM mainly as a result of a higher DM I and milk yield, in comparison to low concentrate diets. Simulation results were evaluated against independent data obtained at three different laboratories in indirect calorimetry trials with COWS consuming GH mainly. The model predicted the average of observed values reasonably, but systematic deviations remained between individual laboratories and root mean squared prediction error was a proportion of 0.12 of the observed mean. Both observed and predicted emission expressed in g CH(4)/kg DM intake decreased upon an increase in dietary N:organic matter (OM) ratio. The model reproduced reasonably well the variation in measured CH, emission in cattle sheds oil Dutch dairy farms and indicated that oil average a fraction of 0.28 of the total emissions must have originated from manure under these circumstances.