Effects of dietary grapeseed extract on performance, energy and nitrogen balance as well as methane and nitrogen losses of lambs and goat kids.

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.

Partitioning of fatty acids into tissues and fluids from reproductive organs of ewes as affected by dietary phenolic extracts.

Evaluation of dietary interventions with regard to fertility problems often observed in ruminant livestock is of global interest. Though the effects of polyphenol supplementation in ruminants on digestion and food quality are well described, the impact on reproductive tissues and fluids remains scarcely investigated. These compounds protect dietary unsaturated fatty acids (FA) from oxidation and biohydrogenation and thus saturation. In addition, modification of the expression of genes associated with FA metabolism may occur. Therefore, we characterized for the first time the FA profiles of reproductive tissues and fluids and investigated their potential modification by dietary polyphenols in 22 cyclic ewes. The animals were randomly divided into four groups and fed a basal diet of meadow hay and one of four concentrate types either non-supplemented (control) or supplemented with grape seed extract, Acacia mearnsii bark extract (13 g/kg dry matter (DM) each) or a combination of both (26 g/kg DM). After 10 weeks of feeding, the animals were slaughtered. Samples of reproductive (oviduct, uterus) and metabolically differently active tissues (liver, muscle, adipose) as well as of plasma and fluids from oviduct and uterus were analysed for their FA composition. In addition, the expression of lipid metabolic and antioxidant genes was analysed in all tissues except the adipose tissue. Fatty acid profiles in tissues and fluids as well as gene expression in tissues significantly differed between the different fluids and tissues. In contrast, only a few diet and matrix (fluid or tissue) × diet interactions were observed. Still, the FA profile of the uterus was the only one not at all affected by the diet. The mRNA expression was not affected by the diet for most of the genes investigated, which might in part be explained by the similar plasma polyphenol concentrations found at slaughter. Overall, our findings contribute to an improved understanding of the characteristic FA composition of reproductive tissues and fluids in sheep. In addition, the effect of polyphenols on different tissues, fluids and tissue gene expression has been confirmed as described in other animal species.

Effect of supplementation of pelleted hazel (Corylus avellana) leaves on blood antioxidant activity, cellular immune response, and heart beat parameters in sheep1.

Hazel leaves (Corylus avellana) fed to sheep resulted in decreased methane emissions without negatively affecting feed intake and were found to have antioxidant properties in vitro. The objective of this study was to evaluate effects of hazel leaves, rich in tannins, on blood antioxidant activity, cellular immune response, and heart beat parameters in sheep. Four experimental pellets were produced by mixing alfalfa and hazel leaves in different proportions, including alfalfa alone as a control, 30% and 60% of hazel leaves, the latter also with 3.8% polyethylene glycol (PEG). Six adult, nonpregnant, nonlactating female sheep (71 ± 5.7 kg of body weight) were allocated to 4 treatments in a 6 × 4 crossover design with four 18-d periods. The diet consisted of experimental pellets and ryegrass-dominated hay (ratio 80% to 20% in dry matter), resulting in hazel leaf proportions of approximately 0%, 25%, and 50% in the total diet. Blood samples were collected at the end of each period to determine plasma total phenol concentration and markers of oxidative status as well as peripheral blood mononuclear cells (PBMC) activation and proliferation response in vitro. Heart rate (HR) and HR variability parameters were measured for 2 consecutive days in each period, during different activities (i.e., eating pellets or hay, or lying). Treatments were compared with multiple comparisons and contrast analysis was used to test for linear and quadratic relations. Compared with control, feeding a high dosage of hazel leaves enhanced (P = 0.006) the plasma total antioxidant capacity, which linearly (P = 0.016) increased with increasing level of hazel leaves in the diet. The total phenol concentration and activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione reductase in the plasma were not different (P ≥ 0.23) among the treatments; however, the latter slightly increased linearly (P = 0.047) with increasing hazel leaves proportion. No differences were observed in the activation and proliferation of PBMC among treatments. The HR decreased linearly (P ≤ 0.009) during pellet eating and lying and the root mean square of successive differences of interbeat intervals (RMSSD) increased linearly (P = 0.037) when lying with increasing level of hazel leaves in the diet. In conclusion, our findings indicate that hazel leaves are a promising supplement to improve oxidative status with no effect on cellular immune response and cardiac stress level of sheep.

Do supplements of Acacia mearnsii and grapeseed extracts alone or in combination alleviate metabolic nitrogen load and manure nitrogen emissions of lambs fed a high crude protein diet?

Diets excessive in crude protein (CP) are unfavourable in terms of metabolic and environmental load. Dietary phenols, often binding to dietary proteins, may alleviate these problems. In an experiment with 60 lambs (3.2 ± 1.6 months of age; 29.7 ± 5.1 kg body weight), kept in pairs, five diets were tested. A diet with 157 g CP/kg dry matter (DM) served as negative control. Four diets with on average 229 (225-233) g CP/kg DM remained either non-supplemented or were supplemented with 13 g/kg DM of Acacia mearnsii extract, grapeseed extract, or a combination of both (26 g extract/kg DM). The analysed concentrations of total extractable phenols were 7.1, 8.1, 14.3, 16.6 and 25.4 g/kg DM for low (CP‒) and high CP (CP+), and high CP with acacia (CP+A), grapeseed (CP+G) and acacia plus grapeseed (CP+AG), respectively. Diets were fed for 10 weeks, and for 6 d faeces and urine were collected and subsequently stored as complete manure for 8 weeks. In blood plasma, phenol concentrations and activities of enzymes indicating liver and kidney stress were analysed. The CP+ diet increased apparent digestibility of N and its removal with the urine, with the expected increase in gaseous N emissions from the manure (13.5 vs 6.5 g/lamb per day during 8 weeks) compared to CP‒. However, no clear signs of metabolic stress were detected. Supplementing the extracts did not impair intake, growth performance and digestibility. Only the supplementation with both extracts decreased urinary N proportion of manure N, and the concomitant weak decline in gaseous emission from the manure was not significant. At least part of the phenols of both extracts seem to be bioavailable as their supplementation elevated blood plasma phenol concentrations by 15% to 40% compared to CP+. A combination of both extracts did not result in a further increase. Further studies have to identify the minimally effective dosage for reducing N emissions, which, at the same time, does not cause adverse side effects in performance.

Transfer of total phenols from a grapeseed-supplemented diet to dairy sheep and goat milk, and effects on performance and milk quality1.

Polyphenols are known to affect digestion of ruminants, whereas there is little information about their metabolic effects. In a 2 × 2-factorial experiment, the effects of supplementing a phenolic grapeseed extract were compared in 11 East Friesian dairy sheep and 9 Saanen goats. The concentrate, supplemented with 7.4 g/100 g DM grapeseed extract, had contents of 3.5 g additional phenols/100 g DM and was compared with a low phenolic control concentrate. Performance, total phenols in blood, milk, urine and feces, antioxidant capacity of the blood, and saliva properties were examined. The experiment lasted for 11 wk from parturition to late lactation, with an initial adaptation phase of 1 wk. Milk yield was measured daily after weaning at about 7 wk after parturition. Blood, milk, saliva, feces, and urine were sampled 4, 3, 2, 2, and 2 times per animal, respectively. The phenolic diet increased phenol concentrations in blood (+10% and 17% in weeks 5 and 11, respectively) and in milk (+32% in week 5) in some of the sampling weeks. There were no clear species differences in phenol concentrations in blood plasma, milk, urine, and feces. However, at the end of the experiment, the supplemented goats had higher (P < 0.05) urinary phenol concentrations than the nonsupplemented goats. A weak relationship (P < 0.05) was found between phenol intake and phenol excretion with milk for sheep but not goats. The phenolic diet did not influence blood antioxidant capacity and tannin-binding capacity of the saliva. The saliva of the goats had a higher tannin-binding capacity than sheep saliva. The effects of the extract on milk yield were inconsistent between sheep and goats. In general, goats had higher feed and nutrient intakes, were heavier, and yielded more milk. Additionally, milk protein and lactose contents were lower and milk urea content was higher in goats than sheep. In conclusion, supplementing grapeseed extract to sheep and goats elevated phenol concentrations in milk and blood to a certain extent, but most of the phenols were lost via urine. The study gave another indication that goats seem to have developed coping mechanisms like a higher salivary tannin-binding capacity, mechanisms which are less pronounced in sheep.

Phenolic plant extracts are additive in their effects against in vitro ruminal methane and ammonia formation.

OBJECTIVE: The methane mitigating potential of various plant-based polyphenol sources is known, but effects of combinations have rarely been tested. The aim of the present study was to determine whether binary and 3-way combinations of such phenol sources affect ruminal fermentation less, similar or more intensively than separate applications. METHODS: The extracts used were from Acacia mearnsii bark (acacia), Vitis vinifera (grape) seed, Camellia sinensis leaves (green tea), Uncaria gambir leaves (gambier), Vaccinium macrocarpon berries (cranberry), Fagopyrum esculentum seed (buckwheat), and Ginkgo biloba leaves (ginkgo). All extracts were tested using the Hohenheim gas test. This was done alone at 5% of dry matter (DM). Acacia was also combined with all other single extracts at 5% of DM each, and with two other phenol sources (all possible combinations) at 2.5%+2.5% of DM. RESULTS: Methane formation was reduced by 7% to 9% by acacia, grape seed and green tea and, in addition, by most extract combinations with acacia. Grape seed and green tea alone and in combination with acacia also reduced methane proportion of total gas to the same degree. The extracts of buckwheat and gingko were poor in phenols and promoted ruminal fermentation. All treatments except green tea alone lowered ammonia concentration by up to 23%, and the binary combinations were more effective as acacia alone. With three extracts, linear effects were found with total gas and methane formation, while with ammonia and other traits linear effects were rare. CONCLUSION: The study identified methane and ammonia mitigating potential of various phenolic plant extracts and showed a number of additive and some non-linear effects of combinations of extracts. Further studies, especially in live animals, should concentrate on combinations of extracts from grape seed, green tea leaves Land acacia bark and determine the ideal dosages of such combinations for the purpose of methane mitigation.

Supplementation of Pelleted Hazel (Corylus avellana) Leaves Decreases Methane and Urinary Nitrogen Emissions by Sheep at Unchanged Forage Intake.

This study is the first to quantify the effects of hazel (Corylus avellana) leaves on methane and urinary nitrogen emissions, digestibility, nitrogen and the energy balance of ruminants. Four experimental pellets were produced with 0, 30% and 60% hazel leaves, the latter also with 4% polyethylene glycol. Hazel leaves gradually replaced lucerne. The diet was composed of the pellets and grass hay (80%: 20%). Six adult sheep were allocated to all four treatments in a 6 × 4 crossover design. Including hazel leaves did not affect the feed intake, but it decreased the apparent digestibility of organic matter and fibre, especially at the high level. Methane emission was reduced by up to 25 to 33% per day, per unit of intake and per unit of organic matter digested. Urinary nitrogen excretion decreased by 33 to 72% with increasing levels of hazel leaves. The treatment with polyethylene glycol demonstrated that tannins in hazel leaves caused significant shares of the effects. In conclusion, the current results indicated a significant potential of hazel leaves as forage for ruminants to mitigate methane and urinary nitrogen emissions. Even high dietary hazel leaf proportions were palatable. The lower digestibility needs to be compensated with easily digestible diet ingredients.

Effects of different stocking densities on performance and activity of cattle × yak hybrids along a transhumance route in the Eastern Himalaya.

Twelve lactating cattle × yak hybrids (B. taurus × B. grunniens) were investigated on five experimental pastures (Sites 1-5), following an up- and downward transhumance route (3,200, 4,000, 4,500, 4,000 and 2,600 m a.s.l.). Hybrids were kept in replicated groups of two (low SD) and four animals per paddock (high SD). As a control, yaks (with calves) were included at low SD at Sites 2-4. Performance was described by body weight, milk yield and composition. Herbage samples as selected by the animals were analyzed. Activity patterns of the hybrids were assessed at Sites 2-4. At similar body weight change and milk composition, the individual hybrids on average produced 26% less milk at high than at low SD. However, at high SD on average still 49% more milk/m(2) of pasture area was produced. Milk fat increased with time from 5.4 to 7.1%, milk protein decreased from 4.2 to 3.3%. At high SD, the hybrids spent less time standing and more time lying. The yaks gained weight at 4,500 m a.s.l. while the hybrids lost weight (+836 and -653 g/animal/day, respectively). In conclusion, high SD reduced individual milk yield and forced hybrids to spend more time for feeding. The yaks apparently had an advantage over the hybrids at very high altitude.

In vitro indications for favourable non-additive effects on ruminal methane mitigation between high-phenolic and high-quality forages.

Feeding plants containing elevated levels of polyphenols may reduce ruminal CH4 emissions, but at the expense of nutrient utilisation. There might, however, be non-additive effects when combining high-phenolic plants with well-digestible, high-nutrient feeds. To test whether non-additive effects exist, the leaves of Carica papaya (high in dietary quality, low in polyphenols), Clidemia hirta (high in hydrolysable tannins), Swietenia mahagoni (high in condensed tannins) and Eugenia aquea (high in non-tannin phenolics) were tested alone and in all possible mixtures (n 15 treatments). An amount of 200 mg DM of samples was incubated in vitro (24 h; 39°C) with buffered rumen fluid using the Hohenheim gas test apparatus. After the incubation, total gas production, CH4 concentration and fermentation profiles were determined. The levels of absolute CH4, and CH4:SCFA and CH4:total gas ratios were lower (P< 0·05) when incubating a combination of C. papaya and any high-phenolic plants (C. hirta, S. mahagoni and E. aquea) than when incubating C. papaya alone. Additionally, mixtures resulted in non-additive effects for all CH4-related parameters of the order of 2-15 % deviation from the expected value (P< 0·01). This means that, by combining these plants, CH4 in relation to the fermentative capacity was lower than that predicted when assuming the linearity of the effects. Similar non-additive effects of combining C. papaya with the other plants were found for NH3 concentrations but not for SCFA concentrations. In conclusion, using mixtures of high-quality plants and high-phenolic plants could be one approach to CH4 mitigation; however, this awaits in vivo confirmation.

Nutrient and energy content, in vitro ruminal fermentation characteristics and methanogenic potential of alpine forage plant species during early summer.

BACKGROUND: Plants growing on alpine meadows are reported to be rich in phenols. Such compounds may affect ruminal fermentation and reduce the plants' methanogenic potential, making alpine grazing advantageous in this respect. The objective of this study was to quantify nutrients and phenols in Alpine forage grasses, herbs and trees collected over 2 years and, in a 24 h in vitro incubation, their effects on ruminal fermentation parameters. RESULTS: The highest in vitro gas production, resulting in metabolisable energy values around 10 MJ kg⁻¹, were found with Alchemilla xanthochlora and Crepis aurea (herbaceous species) and with Sambucus nigra leaves and flowers (tree species). Related to the amount of total gas production, methane formation was highest with Nardus stricta, and lowest with S. nigra and A. xanthochlora. In addition, Castanea sativa leaves led to an exceptional low methane production, but this was accompanied by severely impaired ruminal fermentation. When the data were analysed by principal component analysis, phenol concentrations were negatively related with methane proportion in total gas. CONCLUSION: Variation in methane production potential across the investigated forages was small. The two goals of limited methane production potential and high nutritive value for ruminants were met best by A. xanthochlora and S. nigra.