Tom N Barry

The implications of condensed tannins on the nutritive value of temperate forages fed to ruminants

New methodology for measuring forage condensed tannin (CT) content is described and the effects of CT upon forage feeding and nutritive value for ruminant animals are reviewed. CT react with forage proteins in a pH-reversible manner, with reactivity determined by the concentration, structure and molecular mass of the CT. Increasing concentrations of CT in Lotus corniculatus and Lotus pedunculatus reduce the rates of solubilization and degradation of fraction 1 leaf protein in the rumen and increase duodenal non-NH3 N flow. Action of medium concentrations of total CT in Lotus corniculatus (30-40 g/kg DM) increased the absorption of essential amino acids from the small intestine and increased wool growth, milk secretion and reproductive rate in grazing sheep without affecting voluntary feed intake, thus improving the efficiency of food conversion. High concentrations of CT in Lotus pedunculatus (75-100 g/kg DM) depressed voluntary feed intake and rumen carbohydrate digestion and depressed rates of body and wool growth in grazing sheep. The minimum concentration of CT to prevent rumen frothy bloat in cattle is defined as 5 g/kg DM and sheep grazing CT-containing legumes were shown to better tolerate internal parasite infections than sheep grazing non CT-containing forages. It was concluded that defined concentrations of forage CT can be used to increase the efficiencies of protein digestion and animal productivity in forage-fed ruminants and to develop more ecologically sustainable systems of controlling some diseases under grazing.

Polyphenols and agriculture : beneficial effects of proanthocyanidins in forages

Abstract Proanthocyanidins (PA), also known as condensed tannins, belong to the oldest of plant secondary metabolites. These compounds are widespread in woody plants, but are also found in certain forages. Proanthocyanidins can exert effects in organisms because of their ability to complex with proteins. Forages containing moderate concentrations of PA (2–4% DM) can exert beneficial effects on protein metabolism in sheep, slowing degradation of dietary protein to ammonia by rumen micro-organisms and increasing protein outflow from the rumen, thus increasing absorption of amino acids in the small intestine of the animal. This was shown to result in increases in lactation, wool growth and liveweight gain, without changing voluntary feed intake. Dietary PA can also contribute to improved animal health by reducing the detrimental effects of internal parasites in sheep and the risk of bloat in cattle. In contrast, high dietary PA concentrations (6–12% DM) depress voluntary feed intake, digestive efficiency and animal productivity. Temperate forages containing PA, such as Lotus corniculatus and L. pedunculatus , will not persist in intensive grazing systems if continuously grazed (i.e. set stocked), especially in mixtures with perennial ryegrass and white clover and need to be grown as pure species and rotationally grazed. Nevertheless, inputs of these ‘special purpose’forages can increase sustainability and productivity in intensive grazing systems through increasing the efficiency of animal production, reducing urinary nitrogen (N) excretion and reducing chemical inputs as anthelmintics and as detergents used to control rumen bloat in cattle. Proanthocyanidins are derived from the flavonoid biosynthesis pathway, and knowledge is rapidly increasing about the molecular control of PA biosynthesis. These recent investigations may ultimately enable the expression by genetic engineering of increased levels of PA in the leaves of agriculturally important forage plants such as white clover and perennial rye grass, which will withstand continuous defoliation under grazing and currently contain only trace amounts of PA. This could potentially lead to the widespread use of PA in temperate grazing systems. Further consideration, should now be given to long-term adaptations in the animal, and to ecological effects on the soil ecosystem by PA and their degradation products excreted in animal faeces. More information is required in particular on the effects of PA on soil nitrification, ammonia volatilisation from soil, and nitrogen levels in the groundwater.

The extractable and bound condensed tannin content of leaves from tropical tree, shrub and forage legumes

The extractable, protein-bound and fibre-bound condensed tannin (CT) concentrations in the leaves of tropical legumes grown in both Colombia and Northern Australia were determined by the butanol–HCl method, whilst extractable CT was also determined by the vanillin–HCl method. With the exception of Senna siameaall species contained CT. The very high CT concentration found in many plants growing in Colombia may have been partly due to soil fertility being much lower at the Colombian than the Northern Australian site.Acacia boliviana,Arachis pintoi,Centrosema latidens,Senna velutinaandGliricidia sepiumcontained <55 g total CT kg−1 DM, which suggests that they could comprise a reasonable proportion of ruminant diets. All other species grown in South America contained 100–240 g CT kg−1 DM, which suggests that they should only be fed in small amounts as supplements to dilute the CT concentration.Leucaenaspecies andCalliandra calothyrsusgrown in Northern Australia contained intermediate concentrations of total CT (60–90 g kg−1 DM). Most species contained 70–95% of total CT as extractable CT, with the exception ofFlemingia macrophylla, where 60% was extractable and 40% bound, andGliricidia sepium, where almost all the CT was bound to protein. Values forFlemingia macrophylladiffered between accessions. Extractable CT determined with vanillin–HCl was generally higher than extractable CT determined with butanol–HCl. Three accessions showed negligible (<1 g kg−1 DM) extractable CT with butanol–HCl but 10–12 g extractable CT kg−1 DM with vanillin–HCl. Two accessions showed undetectable levels of extractable CT but substantial levels of protein-bound CT, illustrating the importance of using a bound CT method for identifying forages containing CT. Relative to freeze drying, oven drying ofLeucaenaspecies reduced the concentration of extractable CT and increased concentrations of bound CT. The significance of the results for the nutrition of ruminant livestock are discussed, including the possible roles of protein-bound and fibre-bound CT.

Effects of condensed tannins and crude sesquiterpene lactones extracted from chicory on the motility of larvae of deer lungworm and gastrointestinal nematodes

The objective of this study was to determine the effects of condensed tannins (CT) and an extract containing crude sesquiterpene lactones (CSL) from chicory (Cichorium intybus) on the motility of the first-(L1) and third-stage (L3) larvae of deer lungworm Dictyocaulus viviparus and the L3 larvae of gastrointestinal nematodes in vitro, using the larval migration inhibition (LMI) assay. The CT and CSL had a profound effect on the motility of the larvae displayed by their ability to inhibit larval passage through nylon mesh sieves. Incubation of lungworm L1 larvae in rumen fluid (collected from deer fed pasture) containing 100, 400 and 1000 microg CT/ml, inhibited 12, 28 and 41% of the larvae from passing through the sieves, respectively, while the incubation of L3 larvae with rumen fluid (pH 6.6) containing the same concentrations inhibited 26, 37 and 67% of L3 larvae from passing through the sieves, respectively. Gastrointestinal larvae seem more susceptible to CT than lungworm larvae especially at higher concentrations. CT inhibited 27, 56 and 73% of gastrointestinal larvae from passing through the sieves when used at a concentration of 100, 400 and 1000 microg/ml, respectively. CT were more effective (P<0.001) at reducing the motility of lungworm L1 and L3 larvae when added to the rumen fluid than when added to the abomasal fluid (pH 3.0). Addition of 2 microg polyethylene glycol/microg CT eliminated the inhibitory effect of CT against L1 and L3 larvae especially during incubation in rumen fluid, confirming the effect as due to CT. The CSL extract also showed similar inhibitory activity against L1 and L3 lungworm and L3 gastrointestinal larvae in both fluids, indicating that this extract was not affected by the pH of the fluid, and was more effective against L3 than L1 lungworm larvae. Condensed tannins appeared to be more effective than CSL at inactivating L1 and L3 lungworm and L3 gastrointestinal larvae in rumen fluid, but CSL were particularly effective against L3 lungworm larvae in abomasal fluid. Activity of these secondary compounds explains the reduced parasite problem of young deer grazing chicory.

The efficiency of chewing during eating and ruminating in goats and sheep.

The total amounts of time spent eating and ruminating per 24 h by goats and sheep were determined. The efficiencies of chewing during eating (mean value of C.EAT) and chewing during ruminating (mean value of C.RUM) on the breakdown of feed particles to below the critical size required to leave the rumen (less than 1.0 mm) were investigated. All studies were done with the animals fed on a chaffed lucerne (Medicago sativa) hay diet. Goats spent more time eating (+3.1 h; P less than 0.01), and less time ruminating (-2.2 h; P less than 0.05) per 24 h, than sheep, when fed hourly at ad lib. intake. The efficiency of chewing during eating (mean value of C.EAT) in breaking down feed particles to less than 1.0 mm was greater in goats (85%; P less than 0.01) than sheep (48%). The process of rumination in sheep served to reduce the feed particles which were greater than 1.0 mm in the rumen to less than 1.0 mm. Sheep tended to be more efficient in this process than goats (59 v. 48%), with the difference not attaining significance (P greater than 0.1). The greater frequency of chews (number of total jaw movements/min) during eating in goats (P less than 0.01), or during ruminating in sheep (P less than 0.001), was the major component explaining differences in efficiency between the two species in both the eating and rumination processes. When corrected for the number of chews/min, the differences in mean value of C.RUM and mean value of C.EAT were not significant between goats and sheep.(ABSTRACT TRUNCATED AT 250 WORDS)

Condensed tannins from Lotus corniculatus and Lotus pedunculatus exert different effects on the in vitro rumen degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) protein

Condensed tannins (CT) or proanthocyanidins (PA), which occur in a restricted range of forages, have the ability to interact with proteins and enzymes and can influence the digestion of plant protein in the rumen. We compared the effects of CT extracts from Lotus corniculatus and pedunculatus on degradation of the principal leaf protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), by rumen microorganisms. Total soluble leaf protein extracted from white clover (Trifolium repens ) was incubated with fresh rumen fluid from sheep and a range of concentrations of each CT extract. The rate of degradation of the large (LSU) and small subunit (SSU) of Rubisco was quantified by fractionating the proteins in samples taken from in vitro rumen incubations using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and imaging densitometry. To deduce the effects of the CT extracts, experiments were performed in the presence (CT inactive) and absence (CT active) of polyethylene glycol (PEG; MW 3350). The two CT extracts differed markedly in their effects on the degradation of the LSU and SSU of Rubisco. At concentrations of 0.89 and 1.79 mg CT mg −1 total soluble leaf protein, the CT extract from L pedunculatus was more effective at preventing the degradation of the LSU and SSU by rumen microorganisms than the CT extract from L corniculatus. At a concentration of 1.79 mg CT mg −1 total soluble leaf protein, the CT extracts from L corniculatus and pedunculatus prevented about 0.75 and 0.83 of the LSU and about 0.69 and 0.86 of the SSU, respectively, from being degraded. Addition of PEG removed the inhibition and almost complete degradation of these proteins occurred, as was the case in incubations without CT extracts. The results of this study suggest that the concentration of CT in the diet and the chemical structure which affects the activity of the CT needs to be considered when assessing the effects of CT on protein metabolism in ruminants. n n© 1999 Society of Chemical Industry

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.

Effects of grazing undrenched weaner deer on chicory or perennial ryegrass/white clover pasture on the viability of gastrointestinal nematodes and lungworms

This study determined the in vitro effects on the viability of internal parasites of grazing undrenched weaner deer on either chicory (Cichorium intybus) or perennial ryegrass (Lolium perenne)/white clover (Trifolium repens) pasture. One experiment investigated the hatching and development of gastrointestinal nematode eggs and larvae, and the development and motility of Li lungworm (Dictyocaulus eckerti) larvae, and a second experiment used larval migration inhibition assays to test the viability of LI lungworm larvae extracted from the faeces of weaner deer grazed on either chicory or pasture when they were incubated with rumen and abomasal fluids from fistulated deer also grazing on chicory or pasture. The incubations were undertaken with and without added condensed tannins purified from chicory and with or without polyethylene glycol (PEG) to bind the tannins. Chicory had no effect on the hatching and development of gastrointestinal nematode eggs and larvae. Grazing chicory reduced the number of lungworm larvae developing to the L3 stage, and Li lungworm larvae from the faeces of chicory-grazed deer were less viable in rumen and abomasal fluid than larvae from pasture-grazed animals. Abomasal fluid was significantly (P<0.001) less inhibitory to the migration of Li lungworms than rumen fluid. When the larvae were incubated in rumen and abomasal fluids from chicory-grazed deer, their passage through sieves was significantly (P<0.001) reduced in comparison with when they were incubated in the fluids from pasture-razed deer. Adding condensed tannins to rumen fluid increased the inhibition of the migration of Li lungworm larvae but PEG removed this inhibition; this effect was not observed with abomasal fluid.

Effect of Heat Treatment Upon the Chemical Composition of Cottonseed Meal and Upon the Reactivity of Cottonseed Condensed Tannins

The effects of heat treatment on the chemical composition of cottonseed meal (CSM), with or without the addition of cottonseed hulls (containing condensed tannins ; CT), and upon reactivity of the CT were studied. Heat was applied in a forced draught oven at 100°C for 2 h. Fluorodinitrobenzene (FDNB)-available lysine, free gossypol, extractable- and bound-CT concentrations, in vitro total nitrogen (N) solubility and the in vitro rumen degradation of the two major seed proteins (52 and 48 kDa) present in cottonseed kernel (which does not contain CT) were determined. The reactivity of CT was assessed by determining N solubility and rumen degradation of cottonseed kernel proteins in the presence or absence of polyethylene glycol (PEG ; molecular weight (MW) 3500), which binds and inactivates CT. Heat treatment reduced the concentrations of free gossypol and FDNB-available lysine by small amounts, reduced measurable total CT content by 13%, reduced the solubility of total N, and reduced potential degradability of the 52 and 48 kDa cottonseed storage proteins by mixed rumen microorganisms. Addition of hulls further depressed solubility of total N and ruminal degradation of the two major storage proteins in cottonseed kernel. The action of PEG in vitro indicated that only part of the depression caused by hull addition could be explained by the presence of CT in the hulls, and that the effects of CT upon N solubility and potential degradability in heated CSM were similar to that in unheated CSM. Addition of hulls also substantially reduced FDNB-available lysine. In commercially produced materials, CSM from the Brisbane mill had a lower total CT content, lower N solubility and lower ruminal protein degradation rate than CSM from the Narrabri mill, but a similar level of FDNB-available lysine. Although application of heat inactivated 13% of the total CT, such that it could no longer be extracted and detected with butanol/HCl, it did not seem to change the overall effects produced by CT in reducing N solubility and protein degradation. The effect of hull addition in reducing available lysine has considerable relevance for feeding CSM to monogastric livestock. Interactions involving heat, hulls and CT need to be further studied.

The effect of cottonseed condensed tannins on the ileal digestibility of amino acids in casein and cottonseed kernel.

The effect of adding cottonseed hulls to casein- and cottonseed-kernel-based diets on the apparent and true ileal digestibility of N and amino acids, and the proportion of this effect accounted for by condensed tannin (CT), were determined using the growing rat. Sixty rats were allocated randomly to ten semipurified diets, containing either casein (four diets) or purified unheated solvent-extracted cottonseed kernel (six diets) as the sole protein source, with Cr2O3 added as an indigestible marker. Two of the casein diets contained no hulls whilst the remaining two diets contained 70 g cottonseed hulls/kg. Two of the cottonseed-kernel-based diets contained no hulls, with two containing 23 g hulls/kg and the remaining two containing 46 g hulls/kg. For each pair of diets, PEG was either included or excluded. The effect of CT was quantified by comparing control rats (-PEG; CT acting) with PEG-supplemented rats (+PEG; CT inactivated) at each level of dietary hulls. The rats were given their respective experimental diets for 14 d. Each rat was given the food ad libitum for 10 min hourly from 08.00 to 18.00 hours. On day 14, samples of digesta were collected at death from the terminal 150 mm of ileum at 7 h from the first meal. Apparent and true ileal digestibilities were calculated for DM, N and the individual amino acids. The principal finding was that the inclusion of hulls depressed the apparent and true ileal digestibilities of N and amino acids, but with the response differing between diets. With the casein-based diet the mean apparent and true ileal amino acid digestibilities were significantly depressed from 0.89 and 0.96 to 0.85 and 0.92 respectively, by the inclusion of 70 g hulls/kg in the diet, and addition of PEG then restored these to 0.89 and 0.95. All of the depression could be explained by the CT content of the hulls. However, with the cottonseed-kernel-based diet the responses fell into three categories. The apparent and true ileal digestibilities of the essential amino acids cystine and methionine were not affected by hull addition, ileal digestibilities of leucine, isoleucine, lysine, threonine and valine were markedly depressed by hull addition with approximately 50% of the depression being explained by CT, whilst the ileal digestibilities of histidine, arginine and phenylalanine were depressed by hull addition but little or none of this effect could be explained by CT. Thus the effect of hulls on protein digestion clearly differed with source of protein. With the cottonseed-kernel-based diet it seems that components of the hulls other than CT also depressed the apparent and true ileal digestibilities of N and amino acids. The identity of these components is unknown.