T. N. Barry

Assay and digestion of 14C-labelled condensed tannins in the gastrointestinal tract of sheep

Three experiments were conducted to determine the fate of condensed tannins (CT) during digestion in sheep. CT were measured as extractable, protein-bound and fibre-bound fractions using the butanol-HCl procedure. In Expt 1, purified CT were added to digesta from different parts of the digestive tract obtained from a pasture-fed sheep. Recoveries of CT after 0 and 4 h of anaerobic incubation at 39 degrees averaged: rumen 78.9 and 57.5%; abomasum 50.9 and 49.0%; duodenum 64.4 and 46.0% and ileum 43.4 and 38.8%. In Expt 2, [14C]CT was given per abomasum over a 6.5 h period at 15 min intervals to a sheep previously fed on Lotus pedunculatus (which contains CT). The sheep was killed at the end of the period and 92.4% of the label was recovered. Virtually all of the label was in the digesta, and none was detected in the blood, so that the CT-carbon appeared not to be absorbed from the small intestine. In Expt 3, rumen, abomasal and ileal digesta and faeces samples from sheep fed on Lotus pedunculatus were analysed for CT and CT flow along the digestive tract calculated from reference to indigestible markers. Values were low in all digesta samples, indicating disappearance of CT across the rumen and small intestine, and CT recovery in faeces was only about 15% of intake. However, the 14C results from Expt 2 suggested that little if any CT-carbon was absorbed and the low recoveries in Expt 1 are considered to be a consequence of either conformational changes to the CT molecule such that it is no longer detectable by colorimetric methods, an inability of the analytical method to release bound CT for the butanol-HCl assay, or interference from other digesta constituents. It is concluded that the butanol-HCl method of CT analysis is appropriate for quantifying CT in herbages but not in digesta or faeces, and that a substantial part of CT released during protein digestion in the small intestine may not be detectable by normal CT analytical methods.

Effect of condensed tannins upon the performance of lambs grazing Lotus corniculatus and lucerne ( Medicago sativa )

A grazing experiment, conducted for 22 weeks in 1992/93 at Aorangi Research Station, AgResearch Grasslands, Manawatu, New Zealand, compared the productivity of weaned lambs grazing Lotus corniculatus (birdsfoot trefoil) and lucerne (Medicago sativa). Effects of condensed tannins (CT) in lotus were evaluated by studying the responses of lambs to twice daily oral supplementation with polyethylene glycol (PEG). A rotational grazing system with restricted feed allowance was used. Measurements were made of pre- and post-grazing herbage mass, the composition of the feed on offer and diet selected, voluntary feed intake (VFI), liveweight gain (LWG), carcass growth, wool growth and the concentration of metabolites in rumen fluid. For both lotus and lucerne swards, the diet selected was mainly leaf. Lotus contained 34 g total CT/kg dry matter in the diet selected, whilst there were essentially no CT in lucerne. Compared to lambs grazing lucerne, lambs grazing lotus had slightly lower VFI, and higher LWG, carcass weight gain, carcass dressing-out percentage and wool growth. PEG supplementation had no effect on these measurements or upon the composition of rumen fluid in lambs grazing lucerne. However, in lambs grazing lotus, PEG supplementation reduced wool growth (10.9 v. 12.1 g/day), slightly reduced LWG (188 v. 203 g/day), increased rumen ammonia concentration, and increased the molar proportions of iso-butyric, isovaleric and n-valeric acids and protozoa numbers in rumen fluid. PEG supplementation did not affect carcass gain, carcass fatness or the molar proportion of rumen acetic, propionic or n-butyric acids in lambs grazing lotus. It was concluded that the principal effect of CT in growing lambs grazing lotus was to increase wool growth without affecting VFI, thereby increasing the efficiency of wool production, that the greater rate of carcass gain of lambs grazing lotus than those grazing lucerne was mainly caused by factors other than CT and that CT did not affect the rumen fermentation of carbohydrate to major volatile fatty acids.

Effect of condensed tannins in Lotus corniculatus upon lactation performance in ewes

A grazing experiment was conducted for 8 weeks in the spring/summer of 1993 at Palmerston North, New Zealand, to study the effects of condensed tannins (CT) in Lotus corniculatus (birdsfoot trefoil cv. Grasslands Goldie) upon the lactation performance of ewes rearing twin lambs. Effects of CT were evaluated by studying the responses of ewes to twice daily oral supplementation with polyethylene glycol (PEG; MW 3500), which binds and inactivates CT. A rotational grazing system with restricted feed allowance was used. Measurements were made of pre- and post-grazing herbage mass, the composition of the feed on offer and diet selected, voluntary feed intake (VF1), milk yield and composition, liveweight gain and wool production. The concentration of metabolites in rumen fluid and in blood plasma was also measured. Lotus contained 35–5 g total nitrogen and 44–5 g total CT/kg dry matter in the diet selected, with an in vitro digestibility of 73%. At peak lactation (weeks 3 and 4) milk yield and composition were similar for control (CT-acting) and PEG-supplemented (CTinactivated) ewes but, as lactation progressed, the decline in milk production and in the secretion rates of protein and lactose were less for control than for PEG-supplemented ewes. In mid and late lactation (weeks 6–11), control ewes secreted more milk (21 %), more milk protein (14%) and more lactose (12%) than PEG-supplemented ewes. Milk fat percentage was lower for control than for PEG-supplemented ewes, but secretion rates of fat were similar for the two groups. VFI, liveweight gain and wool growth were similar for both groups. Plasma urea and glucose concentrations were lower for control than for PEG-supplemented ewes, but concentrations of non-esterified fatty acids (NEFA), growth hormone and insulin were similar for the two groups. The concentrations of ammonia and molar proportions of iso -butyric, iso -and n -valeric acids in rumen fluid were lower for control than for PEG-supplemented ewes; molar proportions of acetic, propionic and H-butyric acids were similar for the two groups. It was concluded that for ewes rearing twin lambs grazing L. corniculatus , the action of CT increased milk yield and the secretion rates of protein and lactose without affecting VFI, thereby increasing the efficiency of milk production. The increased milk production did not appear to be mediated by effects on plasma concentrations of growth hormone or insulin.

Effect of condensed tannins upon body growth, wool growth and rumen metabolism in sheep grazing sulla ( Hedysarum coronarium ) and perennial pasture

Areas of sulla ( Hedysarum coronarium ) and pasture ( Lolium perenne/Trifolium repens/Holcus lanatus ) were grazed by young sheep (29·5–34·8 kg initial liveweight) in four experiments, and effects upon body growth, wool growth and rumen metabolism were measured. Sulla contained 40–50 g condensed tannins (CT)/kg DM, whilst the pasture contained small amounts of CT (2–6 g/kg DM). After chewing during eating, a lower proportion of total CT was readily extractable and greater proportions were protein-bound and fibre-bound. Nutritional effects attributable to CT were assessed by oral administration of polyethylene glycol (PEG), which binds and inactivates CT, to half the animals grazing each forage. Rates of body growth were consistently higher for lambs grazing sulla than those grazing pasture, with the CT concentration in sulla being neither stimulatory or inhibitory to body growth or voluntary feed intake (VFI). The action of CT reduced carcass fatness in sheep fed both diets, in the one experiment where this was measured. CT present in both sulla and pasture decreased rumen ammonia concentration and decreased molar proportions of iso-butyrate and iso and n -valerate. During spring and early summer, when wool growth rates were highest, CT present in both pasture and sulla increased wool growth rate; when wool growth rates were low during winter, CT had no effect upon the wool growth of sheep grazing either forage. Numbers of protozoa and molar proportions of n -butyrate in rumen fluid were increased by CT in sheep grazing sulla but not pasture. It was concluded that the higher rates of body growth and VFI in lambs grazing sulla was most likely to be due to its very high ratio of readily fermentable: structural carbohydrate.

The feeding value of chicory (Cichorium intybus) for ruminant livestock

Chicory ( Cichorium intybus ) is perhaps best known for the extract of its roots used as an ingredient in ‘coffee substitute’ beverages. It is less well known as a grazed forage for ruminants. Thomas et al . (1952) reported the high content of some major and minor trace minerals in chicory grown in the UK, and commented on its use in pasture mixtures as a source of these minerals. Chicory was first mentioned in New Zealand (NZ) literature as an animal forage by Cockayne (1915), but a long period then elapsed before Lancashire (1978) reported its excellent value for forage production under rotational grazing in dry summer conditions. Plant selection then followed and the cultivar ‘Grasslands Puna’ was approved for commercial release as a grazed forage plant in 1985 (Rumball 1986). The use of Puna chicory has now spread throughout NZ and the variety is also being used commercially in Australia, North America and South America and is being evaluated in parts of Europe and Asia (W. Green, personal communication). Chicory is a herb, whereas other temperate forages used for ruminant production are either grasses or legumes. This paper reviews work on the chemical composition, nutritive value and feeding value of chicory relative to perennial ryegrass ( Lolium perenne ) and to red clover ( Trifolium pratense ), a legume that, like chicory, is used as a forage for dry summer conditions. Throughout this paper, feeding value is defined as the animal production response to grazing a forage under unrestricted conditions (Ulyatt 1973), with its components being voluntary feed intake (VFI), the digestive process and the efficiency of utilization of digested nutrients; the latter two comprise nutritive value/dry matter (DM) eaten.

Rumen function and digestion parameters associated with differences between sheep in methane emissions when fed chaffed lucerne hay

An indoor experiment involving 10 rumen-cannulated Romney sheep was conducted in May and June 1998 at AgResearch Grasslands, Palmerston North, New Zealand, under restricted feeding conditions, in order to test the hypothesis that animal factors, in particular rumen fractional outflow rate (FOR) and rumen volume, have an influence on the between-sheep variation in methane (CH 4 ) emission. Sheep were fed 2-hourly on chaffed lucerne hay. Following an acclimatization period of 21 days, the experiment lasted 16 days. Energy and nitrogen (N) balances were measured on days 1–6. Cr-EDTA marker was continuously infused into the rumen from day 9 to 16, and rumen contents emptied and sampled on days 13 and 16. Particulate and fluid FOR were estimated using feed lignin and Cr-EDTA, respectively. Daily CH 4 production was measured by the sulphur hexafluoride tracer technique on days 2, 5, 6, 12 and 15 of the experiment. CH 4 production (g/day) was positively correlated with the pool size of organic matter (OM) in the rumen (OM pool, g) ( r =0·84, P =0·002), OM intake (OMI, g/day) ( r =0·67, P =0·04), and the rumen fill (g, wet digesta) ( r =0·76, P =0·01). Multiple regression analysis showed that CH 4 production was best predicted ( R 2 =0·88) as a function of OM pool and the molar % of butyrate; however, OM pool alone accounted for a large proportion ( R 2 =0·71) of the variation in CH 4 production. CH 4 yield (% gross energy intake, % GEI) was negatively correlated with the particulate FOR (%/h) ( r =−0·75, P =0·01) and buffering capacity of rumen fluid (mmol HCl) ( r =−0·72, P =0·02), but positively correlated with the digestibility of cellulose ( r =0·66, P =0·04). Multiple regression analysis showed that CH 4 yield was best predicted as a function of particulate FOR, OMI (g/kg liveweight 0·75 ) and the molar % of butyrate ( R 2 =0·88). Particulate FOR alone explained a large proportion ( R 2 =0·57) of the variation in CH 4 yield. Particulate FOR was negatively correlated with rumen fill ( r =−0·69, P =0·03) and digestibility of cellulose ( r =−0·65, P =0·04). These results suggest that sheep with lower rumen particulate FOR (i.e. longer rumen retention times) had larger rumen fills and higher fibre digestibilities and CH 4 yields. If rumen particulate FOR is to be used as a tool for CH 4 mitigation, the repeatability of its relationship to CH 4 emission must be assessed, preferably under grazing conditions.

The effect of condensed tannins in Lotus pedunculatus on the digestion and metabolism of methionine, cystine and inorganic sulphur in sheep

Two experiments were conducted with sheep fed on fresh Lotus pedunculatus containing 50-55 g condensed tannin (CT)/kg dry matter. Effects of CT were assessed by comparing control sheep (CT operating) with sheep receiving a continuous intraruminal infusion of polyethylene glycol (PEG) to bind and inactivate CT. Digestion of methionine and cystine was determined using a continuous intraruminal infusion of indigestible markers, whilst plasma irreversible loss (IRL) of methionine, cystine and inorganic sulphate was determined using 35S labelling. The proportion of microbial non-NH3-N (NAN) in whole rumen digesta NAN and the IRL of reducible S from the rumen were determined using a continuous intraruminal infusion of (NH4)2(35)SO4. The proportion of microbial NAN in whole rumen digesta NAN (0.44 v. 0.71) and the IRL of reducible S from the rumen (0.84 v. 2.49 g S/d) were lower in control than PEG sheep. PEG sheep lost 30% of ingested methionine and cystine across the rumen, whereas the control sheep lost no methionine and cystine across the rumen. Apparent absorption of methionine from the small intestine was 27% higher in control than PEG sheep, but both groups had a similar apparent absorption of cystine. The apparent digestibility of cystine in the small intestine was lower in control (0.42) than PEG (0.53) sheep, whereas the apparent digestibility of methionine was similar (0.78) for both groups. CT had no effect on plasma methionine IRL, but markedly increased the IRL of cystine (39.8 v. 22.4 mumol/min) and reduced the IRL of plasma inorganic sulphate (35.9 v. 50.2 mumol/min). A three-pool model comparing interconversions between the three plasma metabolites showed that CT increased the flow of cystine to body synthetic reactions (36.5 v. 17.3 mumol/min). This was due to trans-sulphuration of methionine to cystine being greater in control than in PEG sheep, whilst the oxidation of both methionine and cystine were reduced in control sheep. It was concluded that CT reduced the proteolysis of forage protein and the degradation of S amino acids to inorganic sulphide in the rumen, resulting in increased net absorption of methionine and increased utilization of cystine for body synthetic reactions in sheep with a high capacity for wool growth (and, hence, high cystine requirement).

The effect of condensed tannins in Lotus corniculatus on plasma metabolism of methionine, cystine and inorganic sulphate by sheep

Fresh Lotus corniculatus containing 27 g extractable condensed tannin (CT)/kg dry matter (DM) and 8 g bound CT/kg DM was fed at hourly intervals to sheep held in metabolism cages to study the effects of CT on nutrient digestion and on metabolism of methionine, cystine and inorganic sulphate in plasma. Polyethylene glycol (PEG) was continuously infused into the rumen of half the sheep to remove the effects of CT. Principal measurements in the two groups were plasma irreversible loss (IRL) rate and interconversions of methionine, cystine and inorganic sulphate using 35S labelling. CT in Lotus corniculatus had no effects on the apparent digestion of cellulose and minerals, slightly depressed DM, organic matter and hemicellulose digestion and markedly reduced the apparent digestion of N (P < 0.01). The concentration of NH3 and molar proportions of iso-butyric acid, iso-valeric acid and n-valeric acid in rumen fluid were markedly increased by the PEG infusion (P < 0.01), whereas total volatile fatty acid concentration and molar proportions of acetic acid, propionic acid and n-butyric acid were not affected. PEG infusion temporarily increased rumen protozoa numbers. CT greatly increased the IRL of plasma cystine (13.1 v. 7.0 mumol/min; P < 0.05) and reduced IRL of plasma inorganic sulphate (36.8 v. 48.1 mumol/min; P < 0.01) but had no effect on methionine IRL. CT increased transulphuration of methionine to cystine (4.37 v. 1.24 mumol/min; P < 0.05), increased cystine entering the plasma from whole-body protein turnover plus absorption from the small intestine (9.34 v. 5.75 mumol/min; P < 0.05) and increased cystine flux to body synthetic reactions (11.89 v. 5.41 mumol/min; P < 0.05). CT had no effect on the proportion of methionine total flux transferred to sulphate (0.05 v. 0.06; P < 0.05), reduced the proportion of methionine flux transferred to body synthetic reactions (0.68 v. 0.86) and markedly reduced the proportion of cystine flux transferred to sulphate (0.09 v. 0.27; P < 0.01). It was concluded that CT in Lotus corniculatus reduced rumen protein degradation and markedly increased utilization of plasma cystine for body synthetic reactions.

The effect of condensed tannins in Lotus pedunculatus on the solubilization and degradation of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39; Rubisco) protein in the rumen and the sites of Rubisco digestion

Three experiments were undertaken to determine the effect of condensed tannin (CT) in Lotus pedunculatus (45-55 g extractable CT/kg DM) on the digestion of the principal leaf protein, ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39; Rubisco; fraction 1 leaf protein). In two of the experiments Lotus pedunculatus was fed to sheep, with one group receiving a continuous intraruminal infusion (per fistulum) of PEG (molecular weight 3500) to bind and inactivate the CT (PEG group). The other group, which did not receive PEG, was termed the control sheep (CT acting). Expt 3 involved in vitro incubations of Lotus pedunculatus in buffered rumen fluid, with and without PEG added. In all experiments the results have been interpreted in terms of the effects of CT on Rubisco solubilization and degradation. Disappearance of N and Rubisco from Lotus pedunculatus suspended in polyester bags in the rumen was used as a measure of solubilization. Degradation was defined as the disappearance of Rubisco from in vitro incubations of Lotus pedunculatus in rumen fluid. In Expt 1, CT reduced the digestion of Rubisco in the rumen from 0.96 to 0.72 of intake (P < 0.01). Rubisco digestion in the small intestine was 0.27 of intake in control sheep and 0.04 of intake in PEG sheep. In Expt 2, PEG had no effect on the loss of Rubisco from Lotus pedunculatus contained in polyester bags which were incubated in the rumen, hence CT did not affect the solubilization of Rubisco. Observations in Expt 1 were confirmed by in vitro incubations in Expt 3, where PEG addition substantially increased the rate of degradation of plant protein to NH3. Addition of PEG decreased the period of time taken to degrade 50% of the Rubisco from about 13.8 h to about 3.0 h. It was concluded that the action of CT reduced the digestion of Rubisco in the rumen of sheep fed on fresh Lotus pedunculatus, and that this was primarily due to the ability of CT to slow its degradation by rumen micro-organisms, without affecting its solubilization. Both fresh-minced, and freeze-dried and ground lotus were used for in sacco and in vitro incubations; however, fresh-minced lotus was more suitable for the evaluation of protein solubilization and degradation in fresh forages.

The effect of condensed tannins in Lotus corniculatus upon reproductive efficiency and wool production in sheep during late summer and autumn

A grazing experiment, conducted for 55 days (from 4 March to 29 April) in the late summer/autumn of 1997, at Massey University, Palmerston North, New Zealand, compared the reproductive efficiency and wool growth of ewes grazing Lotus corniculatus (birdsfoot trefoil) or perennial ryegrass (Lolium perenne)/white clover ( Trifolium repens ) dominant pasture (pasture). Half the ewes grazing each forage were given daily oral polyethylene glycol (PEG: molecular weight 3500) supplementation to inactivate the condensed tannins (CT) in lotus. A rotational grazing system with 200 mixed age ewes (54·2±0·88 kg/ewe; 50 ewes/treatment) was used. The effect of forage species and PEG supplementation upon voluntary feed intake (VFI), concentration of plasma metabolites, reproductive efficiency, wool production and wool characteristics was measured during two synchronized oestrous cycles. The ewes were restricted to maintenance feeding for the first 12 days of each oestrous cycle and then increased to ad libitum for the 6 days prior to and including ovulation. Lotus contained 17 g total CT/kg dry matter (DM) in the diet selected. There were only trace amounts of total CT in pasture. In vitro organic matter digestibility (OMD) was higher for lotus (0·82 v. 0·74) than for pasture, whilst lotus contained less nitrogen (N; 37·8 v. 44·5 g/kg OM). Mean ovulation rates (OR) for CT-acting and PEG sheep grazing pasture and lotus were respectively 1·33 v. 1·35 and 1·78 v. 1·56, with corresponding lambing percentages being 1·36 v. 1·36 and 1·70 v. 1·42. Fecundity (number of corpora lutea/ewe ovulating) was greater for ewes grazing lotus than pasture ( P <0·01), and tended to be greater for CT-acting than for PEG sheep grazing lotus ( P =0·06). In unsupplemented sheep, ewes grazing lotus had increased plasma concentrations of branched chain amino acids (BCAA; 57%) and essential amino acids (EAA; 52%) compared to ewes grazing pasture. In ewes grazing pasture, PEG administration had no effect on plasma concentrations of urea and free amino acids, VFI, reproductive efficiency and wool production. However, in sheep grazing lotus, plasma concentrations of urea were significantly lower and concentrations of most amino acids were significantly higher for CT-acting than for PEG supplemented ewes (CT not acting); there was no difference in VFI between these two groups. Compared to ewes grazing pasture, ewes grazing lotus had similar VFI but produced more wool with longer staples and thicker fibre diameter, with there being no effect of PEG supplementation. It was concluded that feeding lotus increased the efficiency of both reproduction and wool production without an increase in VFI, and that a possible cause was the action of CT in increasing plasma EAA and especially BCAA concentration.