The Condensed Tannin Content of a Range of Subtropical and Temperate Forages and the Reactivity of Condensed Tannin with Ribulose- 1,5-bis-phosphate Carboxylase (Rubisco) Protein

A series of subtropical grasses and temperate grasses, herbs and legumes were analysed for the presence of extractable and bound condensed tannin (CT) using colorimetric analysis by the butanol–HCl method. Condensed tannins are routinely purified using affinity chromatography with Sephadex LH-20 as a matrix. Therefore, Sephadex LH-20 extracts were further analysed for the presence of CT by ¹³C nuclear magnetic resonance, for anthocyanidin formation after butanol–HCl treatment and for their ability to precipitate ribulose-1,5-bisphosphate carboxylase (Rubisco) protein from lucerne, at pH 7·0. Criteria for the presence or absence of CT were defined. Trace amounts of CT (0·2–2·5 g kg⁻¹ dry matter; DM) were identified and confirmed in summer grass (Digiteria sanguinalis), perennial ryegrass (Lolium perenne) and red clover (Trifolium pretense), with chicory (Chicorium intybus), lucerne (Medicago sativa) and plantain (Plantago lanceolata) identified as probably containing CT. It was concluded that the subtropical grasses kikuyu (Pennisetum clandestinum), paspalum (Paspalum diatatum), smooth witchgrass (Panicum dichotomiflorum) and crowfoot (Eleusine indica) and the temperate grass, Yorkshire fog (Holcus lanatus) probably did not contain CT. Analysis of the extractable fractions by vanillin–HCl gave higher values for CT than analysis by butanol–HCl and wrongly identified some forages as containing trace levels of CT. It was concluded that vanillin–HCl was not specific enough for the detection of trace levels of CT in forages. These results raise the possibility of plant selection programmes to increase the level of CT in grazed forages to approximately 5 g kg⁻¹ DM, the suggested minimum level required to prevent bloat in cattle and to increase wool growth in sheep. It is suggested that this be considered for perennial ryegrass, chicory, red clover and lucerne.     

Effect of condensed tannins prepared from several forages on the in vitro precipitation of ribulose-1,5-bisphosphate carboxylase (Rubisco) protein and its digestion by trypsin (EC 2.4.21.4) and chymotrypsin (EC 2.4.21.1)

A series of in vitro experiments was undertaken to determine the extent to which Sephadex LH-20 treated extracts from a range of temperate forages precipitated ribulose-1,5-bisphosphate carboxylase (Rubisco) and affected the enzymatic hydrolysis of Rubisco protein by trypsin and chymotrypsin at a range of pH values. Rubisco was chosen because it represents the principal dietary protein for ruminants fed fresh forages. Condensed tannins (CT) or proanthocyanidins (PA) are routinely purified by chromatography using Sephadex LH-20 as a matrix. However, these extracts contained non-CT phenolics together with PA so the term ‘CT extract’ was preferred to ‘PA’ to describe the extracts. The in vitro precipitation of Rubisco provided a means to compare the reactivity of the CT extracts. The amount of CT extract required to precipitate all the Rubisco in 10 μg of total soluble leaf protein from white clover (Trifolium repens) when this protein was incubated with CT extracts of Lotus corniculatus, L pedunculatus and sainfoin (Onobrychis viciifolia) was similar, with between 25 and 50 μg of extract required. The CT extract of sulla (Hedysarum coronarium) also precipitated all the Rubisco, however this only occurred with 50 μg of the extract. The CT extract of dock (Rumex obtusifolius) precipitated all the Rubisco when 5 μg of extract or greater was incubated with total soluble leaf protein. However, the differences between the reactivity of all these CT extracts at a range of pH values appeared to be small. Condensed tannin extracts of L corniculatus and L pedunculatus partially inhibited the hydrolysis of Rubisco by trypsin and chymotrypsin to a similar extent, but the extent of the inhibition was affected by pH. The inhibition was greater at pH 6·0 than 7·0, whilst at pH 8·0, CT extracts had little or no affect on trypsin and chymotrypsin. It was concluded that, although the precipitation of Rubisco provided an ideal method for comparing CT extracts, reactivity alone was unlikely to account for the differences in nutritive value that occur with forages containing CT. © 1998 SCI.     

Comparison of the National Research Council-2001 model with the Dutch system (DVE/OEB) in the prediction of nutrient supply to dairy cows from forages

The objective of this study was to compare the DVE/OEB system (DVE = truly absorbed protein in the small intestine; OEB = degraded protein balance) and the NRC-2001 model in the prediction of supply of protein to dairy cows from selected forages: alfalfa (Medicago sativa L. cv. Pioneer and Beaver) and timothy (Phleum pratense L. cv. Climax and Joliette). Comparisons were made in terms of 1) ruminally synthesized microbial CP, 2) truly absorbed protein in the small intestine, and 3) degraded protein balance. In addition, the effects of variety and cutting stage of the selected forages on the potential nutrient supply to dairy cows were also studied. The results showed that the predicted values from the DVE/OEB system and the NRC-2001 model had significant correlations with high R (>0.96) values. However, using the DVE/OEB system, the overall average microbial protein supply based on available energy was 12% higher, and the truly absorbed protein in the small intestine was 15% lower than that predicted by the NRC-2001 model. The difference was also found in the prediction of the degraded protein balances, which was 11% higher based on data from the NRC-2001 model. These differences are due to considerably different factors used in calculations in the two models, although both are based on similar principles. This indicates that a further refinement is needed for a modern protein evaluation and prediction system. In addition, this study showed that the two alfalfa varieties studied (Pioneer vs. Beaver) had no effect, but cutting stage had a profound influence on ruminally synthesized microbial CP (93, 96, 86 g/kg DM at stage of early bud, late bud, and early bloom, respectively) and truly absorbed intestinal protein predicted by the DVE/OEB system (80, 79, 67 g/kg DM at stage of early bud, late bud, and early bloom, respectively). With timothy, both variety (Climax vs. Joliette) and cutting stage had significant impacts on the potential protein supply predicted by both models. The potential protein supply (DVE or MP) to the dairy cow from Climax timothy was higher than that from Joliette timothy (DVE: 46 vs. 32 g/kg DM; MP: 61 vs. 38 g/kg DM). With increasing stage of cutting, the potential protein supply (DVE or MP) was reduced (DVE: 53, 39, 25 g/kg DM; MP: 62, 51, 36 g/kg DM at stage of joint, prebloom head, and full head, respectively).     

Estimation of phosphorus and potassium concentrations in Bermudagrass

Equations were developed to estimate concentrations of phosphorus and potassium for Coastal bermudagrass [Cynodon dactylon, (L.) Pers.] as related to applied nutrient level and harvest interval. Data from several field studies were used in the analysis. Estimates from these agronomic studies agreed with data from fertilizer and waste application for several locations. Concentrations of phosphorus and potassium decreased linearly with harvest interval (up to 6 weeks) and increased exponentially with nutrient level. This information should be relevant to crop production and environmental quality.     

Managing manure nutrients through multi-crop forage production

Concentrated sources of dairy manure represent significant water pollution potential. The southern United States may be more vulnerable to water quality problems than some other regions because of climate, typical farm size, and cropping practices. Dairy manure can be an effective source of plant nutrients and large quantities of nutrients can be recycled through forage production, especially when multi-cropping systems are utilized. Linking forage production with manure utilization is an environmentally sound approach for addressing both of these problems. Review of two triple-crop systems revealed greater N and P recoveries for a corn silage-bermudagrass hay-rye haylage system, whereas forage yields and quality were greater for a corn silage-corn silage-rye haylage system, when manure was applied at rates to supply N. Nutrient uptake was lower than application during the autumn-winter period, and bermudagrass utilized more of the remaining excess than a second crop of corn silage. Economic comparison of these systems suggests that the added value of the two corn silage crop system was not enough to off-set its increased production cost. Therefore, the system that included bermudagrass demonstrated both environmental and economic advantages. Review of the N and P uptake and calculated crop value of various single, double, and triple crop forage systems indicated that the per hectare economic value as well as the N and P uptakes tended to follow DM yields, and grasses tended to out-perform broadleaf forages. Taken across all systems, systems that included bermudagrass tended to have some of the highest economic values and uptakes of N and P. Manure applied at rates to supply N results in application of excess P, and production will not supply adequate quantities of forage to meet the herd's needs. Systems that lower manure application and supply supplemental N to produce all necessary forage under manure application will likely be less economically attractive due to additional costs of moving manure further and, applying it to greater land areas, but will be environmentally necessary in most cases. Intensive forage systems can produce acceptable to high quality forage, protect the environment, and be economically attractive. The optimal manure-forage system will depend on the farm characteristics and specific local conditions. Buffers and nutrient sinks can protect streams and water bodies from migrating nutrients and should be included as a part of crop production systems.     

Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

BACKGROUND: Cellulose and hemicellulose are quantitatively the most important structural carbohydrates present in ruminant diets. Rumen micro‐organisms produce enzymes that catalyse their hydrolysis, but the complex network formed by structural carbohydrates and lignin reduces their digestibility and restricts efficient utilisation of feeds by ruminants. This study aimed to produce two enzymatic extracts, apply them in ruminant diets to determine the best levels for ruminal digestibility and evaluate their effects on in vitro digestibility. RESULTS: In experiment 1 a two‐stage in vitro technique was used to examine the effects of different enzymatic levels of Aspergillus japonicus and Aspergillus terricola on tropical forages. Enzyme addition had minor effects on corn silage at the highest enzymatic level. In experiment 2 an in vitro gas production (GP) technique was applied to determine apparent in vitro organic matter digestibility and metabolisable energy. The addition of enzymes in GP showed interesting results. Good data were obtained using sugar cane and Tifton‐85 hay supplemented with extracts of A. japonicus and A. terricola respectively. CONCLUSION: Overall, the study suggests that addition of crude extracts containing exogenous fibrolytic enzymes to ruminant diets enhances the effective utilisation of ruminant feedstuffs such as forages. Copyright © 2012 Society of Chemical Industry     

Prediction of the nutritive value of forages by near infrared reflectance photometry

A filter-type near infrared photometer equipped with 19 filters was used to predict crude protein, in-vitro digestibility of organic matter, and in-vivo and from in-vitro predicted metabolisable energy in forages. The investigation was performed using pure species of grasses and legumes, grass/legume mixtures and grasses preserved by different methods. The need for specific calibrations is demonstrated. Results obtained using combinations of six filters selected during the calibration procedure were compared with those obtained using the six standard filters recommended for grain analysis. There were indications that selected filter combinations will only give accurate results if the composition of the test samples is very similar to that of the calibration samples, i.e. of the same species or family as the calibration samples. The correlation coefficient between manual methods and n.i.r. reflectance varied from 0.83 to 1.00 for crude protein, 0.48 to 0.91 for rumen digestible organic matter and 0.35 to 0.92 for metabolisable energy. When samples of grass silage were analysed using grass/legume mixtures for calibration of the equipment, the correlations were poor. Although the technique is satisfactory for determinating crude protein in forages utilised in practical farming, the need for specific calibrations when assessing in-vivo organic matter digestibility or in-vivo metabolisable energy renders it inferior to chemical/biological methods.     

Estimation of nitrogen concentration in bermudagrass

Equations were developed to estimate concentration of nitrogen for bermudagrass forage [Cynodon dactylon, (L.) Pers.] as related to applied nitrogen level and harvest interval. Data from six field studies were used in the analysis. Estimates of N concentration in forage obtained with these equations agreed with data from other studies with fertilizer and waste application to bermudagrass (correlation coefficient of 0.94). Concentration of nitrogen decreased linearly with harvest interval (up to 6 weeks) and increased exponentially with nitrogen level. These equations should provide relevant information for crop production and environmental quality.