Garry C. Waghorn

Consequences of plant phenolic compounds for productivity and health of ruminants

Plant phenolic compounds are diverse in structure but are characterised by hydroxylated aromatic rings (e.g. flavan-3-ols). They are categorised as secondary metabolites, and their function in plants is often poorly understood. Many plant phenolic compounds are polymerised into larger molecules such as the proanthocyanidins (PA; condensed tannins) and lignins. Only the lignins, PA, oestrogenic compounds and hydrolysable tannins will be considered here. Lignins slow the physical and microbial degradation of ingested feed, because of resilient covalent bonding with hemicellulose and cellulose, rather than any direct effects on the rumen per se. The PA are prevalent in browse and are expressed in the foliage of some legumes (e.g. Lotus spp.), but rarely in grasses. They reduce the nutritive value of poor-quality diets, but can also have substantial benefits for ruminant productivity and health when improved temperate forages are fed. Beneficial effects are dependent on the chemical and physical structure, and concentration of the PA in the diet, but they have been shown to improve live-weight gain, milk yield and protein concentration, and ovulation rate. They prevent bloat in cattle, reduce gastrointestinal nematode numbers, flystrike and CH4 production. Some phenolic compounds (e.g. coumestans) cause temporary infertility, whilst those produced by Fusarium fungi found in pasture, silage or stored grains can cause permanent infertility. The HT may be toxic because products of their metabolism can cause liver damage and other metabolic disorders.

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 on egg hatching and larval development of Trichostrongylus colubriformis in vitro

The effects of condensed tannins extracted from seven forages on the viability of the eggs and first stage (L1) larvae of the sheep nematode Trichostrongylus colubriformis were evaluated in in vitro assays. The extracts of condensed tannins were obtained from Lotus pedunculatus (LP), Lotus corniculatus (LC), sulla (Hedysarum coronarium), sainfoin (Onobrychus viciifolia), Dorycnium pentaphylum (DP), Dorycnium rectum (DR) and dock (Rumex obtusifolius). Extracts containing 200 to 500,μg/ml reduced the proportion of eggs that hatched. The larval development assay was used to evaluate the effect of the extracts on the development of either eggs or LI larvae to L3 infective larvae. Development was allowed to proceed for seven days by which time the larvae in control incubations had reached the infective L3 stage. Extracts containing 200 μg/mI from LP, DP, DR or dock prevented egg development, and only 11, 8 and 2 per cent of the eggs developed to 13 larvae with extracts from LC, sulla and sainfoin, respectively. When the concentration was 400 μg/mI no eggs developed to L3 larvae. The addition of the extracts after hatching also inhibited the development of Ll to L3 larvae; 200 μg/ml extracted from LP, LC, sulla, sainfoin, DP, DR and dock resulted in only 14, 18, 17, 15, 14, 16 and 4 per cent of Ll larvae developing to the L3 stage compared with 85 per cent for controls, and 400 μg/ml further reduced the development of Li larvae. Statistical analyses showed that when the extracts were added before hatching they were significantly (P<0.001) more effective at inhibiting the larval development than when they were added after hatching. The condensed tannins from dock had the greatest inhibitory effect on egg development followed by the tannins from DR, sainfoin, DP, LP, sulla and LC.

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.