The influence of phenol-rich dietary grapeseed extract on performance, energy and N balance and methane production was determined in sixteen lambs and thirteen goat kids (body weight 20·5 and 19·0 kg, 2 months of age, day 1 of study). Half of the animals received a concentrate containing grapeseed extract, and the others received concentrate without grapeseed extract (total extractable phenols analysed 27 v. 9 g/kg dietary DM; concentrate and hay 1:1). Diets were fed for 7 weeks with 1 week for determining intake, excretion and gaseous exchange in metabolism crates and respiration chambers. Overall, there was an adverse effect of the phenolic diet on apparent N digestibility and body N retention. Faecal N loss as proportion of N intake increased while urinary N loss declined. Relative to N intake, total N excretion was higher and body N retention lower in goat kids than lambs. Diets and animal species had no effect on methane emissions. The saliva of the goat kids had a higher binding capacity for condensed tannins (CT). Goat kids on the phenolic diet had higher CT concentrations in faeces and excreted more CT compared with the lambs (interaction species × diet P < 0·001). The lambs had overall higher (P < 0·001) urinary phenol concentrations than the goat kids (2·19 v. 1·48 g/l). The negative effect on body N retention and lack of effect on methane emissions make the use of the extract in the dosage applied not appealing. Species differences need to be considered in future studies.

The present study investigated the effects of condensed tannins (CT) on intestinal immune function in on-growing grass carp (Ctenopharyngodon idella). A total of 540 healthy grass carp were fed six diets containing different levels of CT (0, 10·00, 20·00, 30·00, 40·00 and 50·00 g/kg diet) for 70 d and then challenged with Aeromonas hydrophila for 14 d. The results showed that, compared with the control group, dietary CT (1) induced intestinal histopathological lesions and aggravated enteritis; (2) decreased lysozyme and acid phosphatase activities, complement 3 (C3), C4 and IgM contents and down-regulated the Hepcidin, liver-expressed antimicrobial peptide (LEAP)-2A, LEAP-2B, Mucin2 and β-defensin-1 mRNA levels in the proximal intestine (PI), mid intestine (MI) and distal intestine (DI) (P < 0·05); (3) down-regulated the mRNA levels of anti-inflammatory cytokines transforming growth factor (TGF)-β1, TGF-β2 (not in MI and DI), IL-4/13A (not IL-4/13B), IL-10 and IL-11 partly correlated with target of rapamycin (TOR) signalling; and (4) up-regulated the mRNA levels of pro-inflammatory cytokines interferon-γ2, IL-1β, IL-6, IL-8 (not in PI), IL-12p35, IL-12p40, IL-15 and IL-17D partly related to NF-κB signalling in the intestine of on-growing grass carp. Overall, the results indicated that CT could impair the intestinal immune function, and its potential regulation mechanisms were partly associated with the TOR and NF-κB signalling pathways. Finally, based on the percentage weight gain and enteritis morbidity, the maximum allowable levels of CT for on-growing grass carp (232·22–890·11 g) were estimated to be 18·6 and 17·4 g/kg diet, respectively.

Currently, consumers are increasingly interested in obtaining high-quality and healthy lamb meat. Compared to grain-based diets, dietary forage legumes such as alfalfa and condensed tannin (CT)-rich sainfoin increase the levels of polyunsaturated fatty acids (PUFAs) that are beneficial for health in lamb meat thanks to their high content in PUFA and/or their impact on ruminal biohydrogenation. However, they can therefore adversely affect its oxidative stability. Thus, the impact of dietary forage legumes on lamb longissimus thoracis (LT) muscle FA composition and their stability to peroxidation was studied in 36 Romane lambs grazing alfalfa (AF; n = 12) or alfalfa plus daily supplementation with CT-rich sainfoin pellets (AS; n = 12; 15 g DM/kg BW, 42 g CT/kg DM) or stall-fed concentrate and grass hay indoors (SI; n = 12). Lambs were slaughtered at a mean age of 162 ± 8.0 days after an average experimental period of 101 ± 8.1 days. Forage legumes-grazing lambs outperformed SI lambs in LT nutritional quality, with more conjugated linoleic acids and n-3 PUFAs, especially 18:3n-3, eicosapentaenoic and docosahexaenoic acids (P < 0.001), and thus lower n-6 PUFA/n-3 PUFA and 18:2 n-6/18:3 n-3 ratios (P < 0.001). Peroxidizability index was higher (P < 0.001) in LT muscle of forage legumes-grazing lambs. Concurrently, two endogenous antioxidant enzyme activities, superoxide dismutase and glutathione peroxidase, were, respectively, similar and lower (P < 0.001) for forage legumes-grazing compared with SI lambs. A lower vitamin E level in SI lambs compared with forage legumes-grazing lambs (1.0 v. 3.8 mg/g, P < 0.001) could explain that malondialdehyde content, a marker of lipid oxidation intensity, was 0.63 µg/g in SI after 8 days in aerobic packaging conditions, whereas it remaining steady at 0.16 µg/g in forage legumes-grazing lambs. Dietary forage alfalfa thus improved FA composition of lamb LT muscle and their stability to oxidation when compared to SI lambs. However, supplementation of alfalfa-grazing lambs with CT-rich sainfoin pellets did not affect the nutritional quality of LT muscle FAs.

Grape marc (GPM) is a viticulture by-product that is rich in secondary compounds, including condensed tannins (CT), and is used as a supplement in livestock feeding practices. The aim of this study was to determine whether feeding GPM to lactating dairy cows would alter the milk proteome through changes in nitrogen (N) partitioning. Ten lactating Holstein cows were fed a total mixed ration (TMR) top-dressed with either 1.5 kg dry matter (DM)/cow/day GPM (GPM group; n = 5) or 2.0 kg DM/cow/day of a 50:50 beet pulp: soy hulls mix (control group; n = 5). Characterization of N partitioning and calculation of N partitioning was completed through analysis of plasma urea-N, urine, feces, and milk urea-N. Milk samples were collected for general composition analysis, HPLC quantification of the high abundance milk proteins (including casein isoforms, α-lactalbumin, and β-lactoglobulin) and liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis of the low abundance protein enriched milk fraction. No differences in DMI, N parameters, or calculated N partitioning were observed across treatments. Dietary treatment did not affect milk yield, milk protein or fat content or yield, or the concentrations of high abundance milk proteins quantified by HPLC analysis. Of the 127 milk proteins that were identified by LC-MS/MS analysis, 16 were affected by treatment, including plasma proteins and proteins associated with the blood-milk barrier, suggesting changes in mammary passage. Immunomodulatory proteins, including butyrophilin subfamily 1 member 1A and serum amyloid A protein, were higher in milk from GPM-fed cows. Heightened abundance of bioactive proteins in milk caused by dietary-induced shifts in mammary passage could be a feasible method to enhance the healthfulness of milk for both the milk-fed calf and human consumer. Additionally, the proteome shifts observed in this trial could provide a starting point for the identification of biomarkers suitable for use as indicators of mammary function.

Little information is available on the effects of different sources of tannins on ruminant product quality. Nowadays several tannin-rich extracts, produced from different plants, are available and contain tannins belonging to different chemical groups, but most of these have not been used so far as feed supplements. The present study aimed at comparing the effects of feeding three tannin extracts (one containing condensed tannins and two containing hydrolysable tannins) to lambs on growth performances and meat oxidative stability. Comisana male lambs were divided into four groups (n=9 each) and were fed for 75 days: a concentrate-based diet (CON), or CON supplemented with 4% tannin extracts from either mimosa (MI; Acacia mearnsii, De Wild; condensed tannins), chestnut (CH; Castanea sativa, Mill; hydrolysable ellagitannins) or tara (TA; Cesalpinia spinosa, (Molina) Kuntze; hydrolysable gallotannins). Only CH reduced growth rate, final weight, carcass weight and feed intake (P<0.05). Tannins did not affect the concentration of the main fatty acid classes and the peroxidability of the intramuscular fat (P>0.05). The TA diet increased (P<0.001) the concentration of γ-tocopherol in muscle and tended to increase that of α-tocopherol (P=0.058). Oxidative stability of raw and cooked meat, or of meat homogenates incubated with pro-oxidants, was not affected by the extracts. These results, compared with those reported in the literature, highlight that some effects of tannins cannot be easily generalized, but may strictly depend on their specific characteristics and on conditions inherent to the basal diet and the metabolic status of the animals.

Previous cafeteria studies suggested that a moderate natural gastrointestinal nematode (GIN) infection did not modify the resource selection of adult Criollo goats towards tannin-rich plants compared with worm-free goats. A higher infection with Haemonchus contortus could trigger a change in the resource selection behaviour towards tannin-rich foliage. Alternatively, goats might select plant species solely to meet their nutritional requirements. A cafeteria study investigated the effect of a high artificial infection with H. contortus on the feed resource selection of goats. Adult Criollo goats (37.5±4.8 kg BW) with browsing experience were distributed in two groups: the infected group (IG) with six animals artificially infected with H. contortus (6000 L3/animal); and the non-infected group (NIG) with six animals maintained worm-free. The experiment included two 5-day periods with additional 5-day adaptation period. In the first period, animals were offered foliage of five plant species with a decreasing gradient of condensed tannins (CT) (Mimosa bahamensis, Gymnopodium floribundum, Havardia albicans, Acacia pennatula, Lysiloma latisiliqum), and three plant species with negligible CT content (Leucaena leucocephala, Piscidia piscipula and Brosimum alicastrum). In the second period the foliage of B. alicastrum was withdrawn. A grain-based concentrate feed was offered daily at 1% BW in DM basis. Dry matter and nutrient intake was determined. Foliage selection of each experimental group was determined using the Chesson selection index. The H. contortus egg count per gram of faeces (EPG) was determined for infected goats twice daily. Chesson index showed a similar pattern of foliage selection on periods 1 and 2. Mean EPG of goats in IG was 2028±259 EPG during period 1 and 1 293±198 EPG during period 2 (P>0.05). During period 1, the selection pattern was highest for B. alicastrum (tannin-free), followed by a tannin-rich plant (M. bahamensis). These two plants remained as highly selected during period 2. The Chesson index showed that both experimental groups (IG and NIG) selected the same plant species in both periods. Thus, a high H. contortus infection did not affect selection of goats fed with CT-rich plants. Apparently, goats balanced their nutrient intake with the plants selected, showing evidence of nutritional wisdom. This balance may have helped to prevent excess protein in the diet and also to maintain a low GIN infection, both considered as examples of prophylactic self-medication.

Many plant tissues contain plant secondary compounds (PSC), which have long been recognised as defensive chemicals that deter herbivory via their toxic effects. However, herbivores may also benefit from including PSC into their diets. Plant-derived phenolics, terpenes and alkaloids have antiparasitic properties and sesquiterpene lactones have antibacterial, antifungal and antiparasitic properties. These actions are in part a consequence of the negative actions that PSC exert across several trophic levels, including the bacteria, parasites and fungi that inhabit herbivores’ bodies. Given the dual action, toxin and medicine, it is possible to hypothesise that self-selection of PSC by herbivores should occur when the benefits outweigh the costs of PSC ingestion. Recent research suggests that sheep and goats self-medicate against parasitic infections. They increase preference for condensed tannin-containing foods when experiencing a parasitic burden. This behaviour improves health; it is triggered by parasitism and weakens when parasitism subsides. However, the causes underlying these responses are not straightforward when viewed under a unidimensional cost–benefit analysis. This is because the intensity of antinutritional/toxic and medicinal effects of PSC is not static or just dependent upon the isolated post-ingestive effects of single PSC. Nutrient–PSC and PSC–PSC interactions, social models, as well as feeding patterns, all influence the perceived net benefit of incorporating medicines into a diet. A better understanding of the net benefit of self-medication in complex feeding environments will allow for the development of innovative managing strategies aimed at providing the food alternatives and conditions for improving the nutrition, health and welfare of grazing animals.

Enteric methane (CH4) emissions from ruminants have compelled a wide range of research initiatives to identify environmental abatement opportunities. However, although such mitigations can theoretically be attained with feed additives and feeding strategies, the limited empirical evidence on plant extracts used as feed additives does not support extensive or long-term reductions. Nevertheless, their strategic use (i.e. alone or combined in a simultaneous or consecutive use) may provide not only acceptable CH4 abatement levels, but also relevant effects on animal physiology and productivity. Condensed tannins (CT) represent a range of polyphenolic compounds of flavan-3-ol units present in some forage species that can also be added to prepared diets. Methods to determine CT, or their conjugated metabolites, are not simple. Although there are limitations and uncertainties about the methods to be applied, CT are thought to reduce CH4 production (1) indirectly by binding to the dietary fibre and/or reducing the rumen digestion and digestibility of the fibre and (2) directly by inhibiting the growth of rumen methanogens. On the basis of their role in livestock nutrition, CT influence the digestion of protein in the rumen because of their affinity for proteins (e.g. oxidative coupling and H bonding at neutral pH) that causes the CT-protein complex to be insoluble in the rumen; and dissociate in the abomasum at pH 2.5 to 3.0 for proteolysis and absorption in the small intestine. CT may also reduce gastro-intestinal parasite burdens and improve reproductive performance, foetal development, immune system response, hormone serum concentrations, wool production and lactation. The objectives of this paper are to discuss some of the beneficial and detrimental effects of CT on ruminant production systems and to develop a conceptual model to illustrate these metabolic relationships in terms of systemic physiology using earlier investigations with the CT-containing legume Lotus corniculatus. Our conceptual model indicated four complex and long-lasting relationships (digestive, toxicological, physiological and morphological) that can alter the normal biology of the animal. These relationships are interdependent, integrative, and sometimes, complementary to each other. This conceptual model can be used to develop mechanistic models to improve the understanding of the interaction between CT and the ruminants as well as to guide research initiatives of the impact of polyphenol-rich foods on human health.

The aim of this study was to investigate whether the use of sainfoin-based condensed tannins (CT) enhances feed value when given with tannin-free legumes (lucerne) to sheep. The experiments were conducted with fresh sainfoin and lucerne harvested at two stages (vegetative stage as compared with early flowering) in the first growth cycle. Fresh sainfoin and lucerne forages were combined in ratios of 100 : 0, 75 : 25, 25 : 75 and 0 : 100 (denoted S100, S75, S25 and S0, respectively). Voluntary intake, organic matter digestibility (OMD) and nitrogen (N) retention were measured in sheep fed the different sainfoin and lucerne mixtures. Loss of dry matter (DM) and N from polyester bags suspended in the rumen, abomasum and small intestine (SI) was also measured using rumen-fistulated sheep and intestinally fistulated sheep. The CT content in sainfoin (S100) decreased with increasing percentage of lucerne in the mixture (mean value from 58 g/kg DM for S100 to 18 g/kg DM for S25) and with growth stage (S100: 64 to 52 g/kg DM). OMD did not differ between different sainfoin/lucerne mixture ratios. Sainfoin and lucerne had an associative effect (significant quadratic contrast) on voluntary intake, N intake, total-tract N digestibility, N in faeces and urine (g/g N intake) and N retained (g/g N intake). Compared with lucerne mixtures (S0 and S25), high-sainfoin-content mixtures (S100 and S75) increased the in situ estimates of forage N escaping from the rumen (from 0.162, 0.188 for S0 and S25 to 0.257, 0.287 for S75 and S100) but decreased forage N intestinal digestibility (from 0.496, 0.446 for S0 and S25 to 0.469, 0.335 for S75 and S100). The amount of forage N disappearing from the bags in the SI (per g forage N) was the highest for high-sainfoin mixtures (from 0.082, 0.108 for S100 and S75 to 0.056, 0.058 for S25 and S0, P < 0.001). Rumen juice total N (tN) and ammonia N (NH3-N) values were the lowest in the high-sainfoin diet (mean tN 0.166 mg/g in S100 as compared with 0.514 mg/g in S0; mean NH3-N 0.104 mg/g in S100 as compared with 0.333 mg/g in S0, P < 0.001).

The nutritional effects of sulla (Hedysarum coronarium L.) forage containing condensed tannins (CT) on growth of lambs, and carcass and meat quality were investigated. Thirty-two male Comisana lambs aged 100 ± 8 days weighing 19.0 ± 2.8 kg were fed fresh forage of sulla or CT-free annual ryegrass (Lolium multiflorum Lam. subsp. Wersterwoldicum) for 49 days until slaughter; in addition, each lamb was supplied with 200 g/days of concentrate. Eight lambs per diet had been previously treated with anthelmintic drugs to remove nematode parasites. Measurements of BW and feed intake, and counts of faecal nematode eggs were made. Carcass parameters were recorded after slaughter, and tissue components of the hind leg were determined. Longissimus dorsi meat was evaluated for pH, colour, thawing and cooking losses, Warner-Bratzler shear force, chemical composition and sensory properties based on triangle tests. Relative to ryegrass-fed lambs, sulla-fed lambs had significantly greater dry matter (DM) and protein intake, a more favourable feed conversion ratio, and superior growth rate, final BW at 150 days of age, carcass weight, yield and fatness. These results were attributed to the high protein and non-structural carbohydrate content of sulla, and also to the moderate CT content of sulla (16.7 and 20.3 g/kg of DM in offered and consumed sulla forage, respectively). Anthelmintic treatment did not affect lamb growth, as the level of parasitic infection (initial and final) was low. The physical, chemical and sensory properties of the lamb meat were not influenced by diet.

Tanniferous forage plants can have beneficial effects on ruminant productivity and health (improved protein supply, bloat safety and antiparasitic properties). However, condensed tannins can also lower palatability, voluntary feed intake and digestibility. The aim of our interdisciplinary project was to generate basic knowledge on plant management, feed palatability and the antiparasitic properties of tanniferous forage plants for their practical application in agronomy, focusing on their usefulness in controlling gastrointestinal nematodes in organic farming. We found that Onobrychis viciifolia (sainfoin), Lotus corniculatus (birdsfoot trefoil) and Cichorium intybus (chicory) were suitable for cultivation under the given temperate climatic conditions, whereas Lotus pedunculatus (big trefoil) was soon outcompeted by unsown species. Growing the tanniferous plant species in a mixture with Festuca pratensis (meadow fescue) rather than in a monoculture had the advantage of increasing total dry matter (DM) yield (especially in the case of tanniferous legumes) and of reducing the DM proportions of unsown species. However, due to dilution by non-tanniferous F. pratensis, the tannin concentrations of mixtures were clearly lower and the seasonal fluctuations in tannin concentrations greater than that of monocultures. Across species, tannin concentrations were highest for O. viciifolia, followed by L. corniculatus and very low for C. intybus. Palatability of all tanniferous forages was comparable to that of a ryegrass/clover mixture when fed as dried forage and, when offered as silage, palatability of O. viciifolia was clearly superior to that of the respective ryegrass/clover control. Administration of dried or ensiled O. viciifolia reduced parasite egg counts in feces of lambs co-infected with the gastrointestinal nematode species Haemonchus contortus and Cooperia curticei. We conclude that O. viciifolia is the most promising among the tested tanniferous forage plant species due to its suitability for cultivation, its high tannin concentration, its high palatability and its antiparasitic activity even in dried or ensiled form.

Flavanols are the most abundant flavonoids in the human diet where they exist as monomers, oligomers and polymers. In the present study, catechin, the procyanidin dimer B3 and a grapeseed extract containing catechin, epicatechin and a mixture of procyanidins were fed to rats in a single meal. After the meals, catechin and epicatechin were present in conjugated forms in both plasma and urine. In contrast, no procyanidins or conjugates were detected in the plasma or urine of any rats. Procyanidins were not cleaved into bioavailable monomers and had no significant effects on the plasma levels or urinary excretion of the monomers when supplied together in the grapeseed extract. We conclude that the nutritional effects of dietary procyanidins are unlikely to be due to procyanidins themselves or monomeric metabolites with the intact flavonoid-ring structure, as they do not exist at detectable concentrations in vivo. Future research should focus on other procyanidin metabolites such as phenolic acids and on the effects of the unabsorbed oligomers and polymers on the human gastrointestinal tract.

A factorial experiment was conducted to study the effects of condensed tannins (CT) from the tropical legumes Desmodium intorturn and Calliandra calothyrsus on the digestion and utilization of protein and carbohydrate in sheep and goats. CT-free Centrusema pubescens was also fed for comparison with the CT legumes, and each legume was included (300 g/kg DM) in a basal diet of pangola grass (Digitmia decumbens). Pangola grass alone was used as a control diet. There were no significant (P>0.05) differences between sheep and goats for the efficiency of digestion of N (0.574, SE 0.013), organic matter (OM; 0.519, SE 0.010), neutral-detergent fibre (NDF; 0.524, SE 0.011) and acid-detergent fibre (ADF; 0.407, SE 0.016). Diets containing desmodium and calliandra were digested less well in the rumen (64 and 62% of total OM digested) when compared with the pangola and centrosema diets (74 and 73% of total OM digested in rumen). There was an apparent net gain of 30% in ADF across the digestive tract of sheep and goats given calliandra, and this gain was ascribed to the formation of ‘artifact’ fibre as a result of fibre-tannin interaction. Overall, inclusion of legume at 300 g/kg in the diet significantly increased (P>0.05) the concentration of acetic acid and decreased butyric acid concentration in the rumen fluid of sheep and goats. Significantly higher proportions of dietary N apparently reached the abomasum of animals given the diets containing desmodium (50%) and calliandra (56%) when compared with animals given the centrosema and pangola diets (35%). Sheep and goats given the CT diets also had higher excretions of faecal N. This increment of faecal N (14%) did not affect post-rumen N digestion (P>0.05) since animals given CT diets absorbed more N (19%) per kg total OM digested than those given the control diets. It was concluded that whilst the low levels of CT provided in desmodium (1.0%) and calliandra (2.3%) diets protected dietary protein from degradation in the rumen, there were no overall beneficial or detrimental effects of CT in these diets for sheep or goats. A method was developed to categorize CT into fractions representative of their forms (free, protein-bound, and fibre-bound) during the digestion process. A quantitative model of CT metabolism during passage through the digestive tract was developed from the measured exchanges of CT between free, protein-bound and fibre-bound pools in the rumen and lower digestive tract. CT interchange mainly occurred in the reticulo-rumen of both animal species. Desmodium and calliandra free CT showed net losses of 68 and 78% in the rumen respectively and 57 and 68% of the fibre-bound CT was lost in the same site for sheep and goats respectively. However, protein-bound CT increased across the rumen by 73 and 56% for both animal species. Post-rumen losses of the total CT abomasal flow were 86 and 83% (free CT) for sheep and goats respectively, 70 and 66% (protein-bound CT), whilst 28% loss of fibre-bound CT occurred in sheep and goats respectively.

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 (131·1 v. 7·0 μmol/min; P 0·05) and reduced IRL of plasma inorganic sulphate (36·8 v. 48·1 μmol/min; P 0·01) but had no effect on methionine IRL. CT increased transulphuration of methionine to cystine (4·37 v. 1·24 μmol/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 μmol/min; P 0·05) and increased cystine flux to body synthetic reactions (11·89 v. 5·41 μmol/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.

Proanthocyanidins (condensed tannins; PA) purified from the leaves of forage legumes Trifolium arvense, Lotus pedunculatus, Lotus corniculatus, Dorycnium rectum, Coronilla varia, Onobrychis viciifolia, or Hedysarum coronarium, were added to soluble lucerne (Medicago sativa) leaf protein and incubated with strained rumen fluid in vitro. Fractions were collected and frozen immediately. Denatured proteins were fractionated by sodium dodecylsulphate–polyacrylamide gel electrophoresis (SDS–PAGE), stained, and relative levels were quantified by densitometry. In the absence of PA the large subunit (LSU) of ribulose bisphosphate carboxylase (EC 4. 1. 1. 39) was susceptible to proteolysis by rumen microflora but the small subunit (SSU) resisted breakdown. PA purified from Onobrychis was added to soluble leaf protein, at PA: protein ratios between 1:1 and 1:20. The rate of proteolysis of LSU1 was significantly reduced at PA: protein ratios of 1:2 and 1:1 (P < 0.001) and the rate of digestion was reduced by between 3- and 21-fold. In separate experiments PA isolated from the range of species described was added to rumen fluid to give PA: protein ratios of 1: 5. The addition of PA significantly reduced the rate of proteolysis of LSU, when compared with PA-free control. There were only small differences between PA from different species. The inhibitory effect of PA may have been due to PA binding to the dietary protein or to the rumen proteases, interfering with the action of proteases on susceptible sites within the substrate.

The effects of condensed tannins in hulls of faba beans (Vicia faba L.) on the activities of trypsin (EC3.4.21.4) and chymotrypsin (EC3.4.21.1) in digesta obtained from the small intestine of pigs were studied. Using four castrated male pigs (mean body weight 83 kg) fitted with both a simple T-cannuIa in the duodenum and a post-valvular T-cannula at the terminal ileum, two experimental diets were tested in a Latin square design. The low-tannin diet (LT) contained 200 g faba bean hulls (cv. Blandine)/kg with a low content of condensed tannins (< 0.1 % catechin equivalents). The high-tannin diet (HT) contained 200 g faba bean hulls (cv. Alfred)/kg with a content of condensed tannins of 3.5 % catechin equivalents. Spot samples of fresh duodenal digesta were taken daily at fifteen time points between 08.00 and 20.00 hours on four consecutive days. Ileal digesta were collected nearly quantitatively on the same days between 08.00 and 20.00 hours over periods of 2 h. Trypsin and chymotrypsin activities in duodenal digesta did not differ between treatments at any time point (P > 0.05). In ileal digesta of pigs given diet HT the mean activity of trypsin was reduced (P < 0.05). The activity of chymotrypsin in ileal digesta did not differ between treatments. Trypsin activity: chymotrypsin activity was somewhat lower in ileal digesta of pigs receiving the HT diet (P < 0.10). The apparent ileal digestibility of crude protein (N × 6.25) was lower for the HT than for the LT diet (0.614 v. 0.728; P < 0.05). Condensed tannins are probably responsible for the lower activity of trypsin in ileal digesta of pigs fed on high-tannin faba bean hulls. Various explanations for the absence of effects of condensed tannins on enzyme activity in duodenal digesta are discussed.

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)235SO4. 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 μmol/min) and reduced the IRL of plasma inorganic sulphate (35·9 v. 50·2 μmol/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 μmol/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).

Seed samples from two near-isogenic faba bean (Vicia faba L.) lines were examined for the levels of so-called anti-nutritional factors (ANF). From the ANF known to be present in faba beans, trypsin inhibitor activity, functional lectins, condensed tannins and pyrimidine glycosides were analysed. It was concluded that the lines differed only in the content of condensed tannins being < 0.5 g/kg and 5.2 (sd 0.2) g/kg for the low (LT)- and high (HT)-tannin lines respectively. In addition, the level of pyrimidine glycosides in the LT line was slightly higher than that in the HT line. The LT line showed a reduced proportion of the seed coat (105 v. 119 g/kg) and a lower seed weight (0.85 v. 1.01 g). The apparent ileal and faecal digestibility values of dry matter and nitrogen from the HT and LT line were determined for piglets which were fed on diets containing chromium oxide as a marker. The mean apparent ileal and faecal digestibility values for dry matter for the LT line were 0.694 and 0.889 and for N 0.828 and 0.879 respectively. For the HT line, these values were approximately 0.05 and 0.10 lower (P > 0.05). A multi-enzyme technique was used to predict the in vitro protein digestibility (IVPD) of the two lines. The IVPD of the LT line was 0.965 and about 0.05 higher than the HT line, confirming the difference in digestibility as measured in vivo. The present study shows the positive effects on digestibility of removal of condensed tannins in faba beans which was achieved by plant breeding.