Effect of dietary tannin supplementation on cow milk quality in two different grazing seasons.

Extensive farming systems are characterized by seasons with different diet quality along the year, as pasture availability is strictly depending on climatic conditions. A number of problems for cattle may occur in each season. Tannins are natural polyphenolic compounds that can be integrated in cows' diet to overcome these seasonal problems, but little is known about their effect on milk quality according to the season. This study was designed to assess the effects of 150 g/head × day of tannin extract supplementation on proximate composition, urea, colour, cheesemaking aptitude, antioxidant capacity, and fatty acid (FA) profile of cow milk, measured during the wet season (WS) and the dry season (DS) of Mediterranean climate. In WS, dietary tannins had marginal effect on milk quality. Conversely, in DS, the milk from cows eating tannins showed 10% lower urea and slight improvement in antioxidant capacity, measured with FRAP and TEAC assays. Also, tannin extract supplementation in DS reduced branched-chain FA concentration, C18:1 t10 to C18:1 t11 ratio and rumenic to linoleic acid ratio. Tannins effect on rumen metabolism was enhanced in the season in which green herbage was not available, probably because of the low protein content, and high acid detergent fibre and lignin contents in diet. Thus, the integration of tannin in the diet should be adapted to the season. This could have practical implications for a more conscious use of tannin-rich extracts, and other tannin sources such as agro-industrial by-products and forages.

Cheese quality from cows given a tannin extract in 2 different grazing seasons.

The aim of the present study was to compare the effect of dietary tannins on cow cheese quality in 2 different grazing seasons in the Mediterranean. Two experiments were performed on 14 dairy cows reared in an extensive system. The first experiment took place in the wet season (WS), and the second experiment took place in the dry season (DS). In the WS and DS experiments, cows freely grazed green pasture or dry stubbles, respectively, and the diet was supplemented with pelleted concentrate and hay. In both experiments, the cows were divided into 2 balanced groups: a control group and a group (TAN) receiving 150 g of tannin extract/head per day. After 23 d of dietary treatment, individual milk was collected, processed into individual cheeses, and aged 25 d. Milk was analyzed for chemical composition, color parameters, and cheesemaking aptitude (laboratory cheese yield and milk coagulation properties). Cheese was analyzed for chemical composition, proteolysis, color parameters, rheological parameters, fatty acid profile, and odor-active volatile compounds. Data from the WS and DS experiments were statistically analyzed separately with an analysis of covariance model. In the WS experiment, dietary tannin supplementation had no effect on milk and cheese parameters except for a reduced concentration of 2-heptanone in cheese. In the DS experiment, TAN milk showed lower urea N, and TAN cheese had lower C18:1 trans-10 concentration and n-6:n-3 polyunsaturated fatty acid ratio compared with the control group. These differences are likely due to the effect of tannins on rumen N metabolism and fatty acid biohydrogenation. Dietary tannins may differently affect the quality of cheese from Mediterranean grazing cows according to the grazing season. Indeed, tannin bioactivity on rumen metabolism seems to be enhanced during the dry season, when diet is low in protein and rich in acid detergent fiber and lignin. The supplementation dose used in this study (1% of estimated dry matter intake) had no detrimental effects on cheese yield or cheesemaking parameters. Also, it is unlikely that sensorial characteristics would be affected by this kind of dietary tannin supplementation.

Effect of increasing the proportion of chicory in forage-based diets on intake and digestion by sheep.

There is a lot of evidence that chicory could be a highly palatable and nutritious source of forage for ruminants, well adapted to climate change and dry conditions in summer, thanks to its resistance to drought and high water content. This study aimed to describe the effect of incorporating chicory to ryegrass or to a ryegrass-white clover mixture on feeding behaviour, digestive parameters, nitrogen (N) balance and methane (CH4) emissions in sheep. In total, three swards of ryegrass, white clover and chicory were established and managed in a manner ensuring the forage use at a constant vegetative stage throughout the experiment. In all, four dietary treatments (pure ryegrass; binary mixture: 50% ryegrass-50% chicory; ternary mixture: 50% ryegrass-25% white clover-25% chicory; and pure chicory) were evaluated in a 4×4 replicated Latin square design with eight young castrated Texel sheep. Each experimental period consisted of an 8-day diet adaptation phase, followed by a 6-day measuring phase during which intake dynamics, chewing activity, digestibility, rumen liquid passage rate, fermentation end-products, N balance and CH4 emissions were determined. Data were analysed using a mixed model and orthogonal contrasts were used to detect the potential associative effects between ryegrass and chicory. The daily voluntary dry matter intake was lower for pure ryegrass than for diets containing chicory (P<0.001) and increased quadratically from 1.39 to 1.74 kg/day with increasing proportion of chicory. Huge positive quadratic effects (P<0.001) between ryegrass and chicory were detected on eating time and eating rate just after feeding indicating an increase of the motivation to eat with mixtures, whereas rumination activity decreased linearly with the proportion of chicory (P<0.001). The organic matter digestibility was similar among treatments (around 80%), but a strong positive quadratic P<0.001) effect was observed on liquid passage rate suggesting that chicory allowed fast particle breakdown in the rumen. Animals fed with the ryegrass-white clover-chicory mixture had the higher urinary N losses (P<0.001), whereas retained N per day or per g N intake was greater when the proportion of chicory was at least 50% (P<0.001) being ~40% greater than for the other treatments. The CH4 yield was lower with pure chicory than with the other treatments (P<0.001) for which emissions were similar. In conclusion, mixing ryegrass and chicory in equal proportions produces a synergy on voluntary intake and an improved N use efficiency likely due to complementarity in chemical composition, increased motivation to eat and faster ruminal particle breakdown.

Fatty acid composition of ruminal digesta and longissimus muscle from lambs fed silage mixtures including red clover, sainfoin, and timothy.

This work investigated the effects of feeding silage mixtures of a plant containing polyphenol oxidase (PPO; red clover [; RC]), a plant containing tannins (sainfoin [; SF]), and a grass species not containing these compounds (timothy [; T]) on ruminal and intramuscular (i.m.) fatty acids of lambs. Forty 4-mo-old castrated male Romane lambs, divided into 5 groups, received 1 of the following silages: 1) T (100%), 2) a binary mixture of timothy and tannin-containing sainfoin ( cv. Perly; 50:50 [T-SF]), 3) a binary mixture of timothy and PPO-containing red clover ( cv. Mervius; 50:50 [T-RC]), 4) a ternary mixture of timothy, sainfoin, and red clover containing both tannins and PPO (50:25:25, respectively [T-SF-RC]), and 5) a binary mixture of tannin-containing sainfoin and PPO-containing red clover (50:50 [SF-RC]). In the rumen digesta, the partial or total replacement of T with forage legumes was associated with greater concentrations of PUFA ( < 0.001) and 1esser concentrations of MUFA ( < 0.001). The inclusion of forage legumes in the silage favored the accumulation of 18:3 -3 ( < 0.001), with the greatest concentrations being observed in SF-RC. This latter diet also led to the greatest percentage of 18:2 -6 ( < 0.001). Forage legumes decreased the -11 18:1 to 30% of T in rumen digesta ( < 0.001). Forage legumes decreased the total concentration of branched-chain fatty acids in the rumen digesta (on average, -28%; < 0.001), this effect being less marked (-17%; = 0.014) in T-RC in comparison with T. The dietary treatment tended to affect the proportion of MUFA ( = 0.081) and of PUFA ( = 0.079) in the i.m. fat of the LM, respectively, at the highest and lowest numerical value in the T group. The sum of -3 fatty acids was less in the T and T-SF groups compared with the mixture of legumes without T (SF-RC; < 0.001 and < 0.008, respectively). The latter group had also a lesser -6-to--3 ratio than the T-SF group ( = 0.01). -11 18:1 was greater ( < 0.03) in lambs given T as the sole silage in comparison with lambs given T-RC, T-SF-RC, and SF-RC. Rumenic acid (-9 -11 CLA) was detected at a greater percentage in the LM from the animals fed the T silage compared with animals fed the T-SF-RC treatment ( = 0.004). Contrarily, -9 -12 -15 18:3 was found at a greater concentration in the muscle from lambs in the SF-RC treatment compared with lambs in the other treatments ( < 0.001). Furthermore, the T-RC group showed a greater proportion of α-linolenic acid than the T group ( = 0.03). In conclusion, the effects of the 2 active compounds (PPO and tannins) appear to be similar and additive. Moreover, a silage composed of a mixture of RC and SF is an excellent forage for growing lambs in terms of i.m. fatty acid composition.

Associative effects between orchardgrass and red clover silages on voluntary intake and digestion in sheep: Evidence of a synergy on digestible dry matter intake.

The aim of this study was to assess the associative effects between orchardgrass () and red clover () silages as a model of preserved grass-legume mixture on voluntary intake parameters and digestive efficiency in sheep. Ten sheep were assigned to a repeated 5 × 5 Latin square design, in which 5 proportions of orchardgrass and red clover silages were tested (0:100, 25:75, 50:50, 75:25, and 0:100, on a DM basis). Measurements were performed simultaneously on intake, feeding behavior (eating time and chewing activity), digestive parameters (nutrient digestibility and rumen parameters), nitrogen balance, and enteric methane (CH) emissions using the SF6 tracer technique. Positive associative effects were observed on daily voluntary DMI ( < 0.001), which was greater when sheep were fed with at least 50% red clover (1.56-1.59 kg/d) compared with those fed with 0 or 25% red clover (1.29 and 1.45 kg/d, respectively; < 0.001). This synergy between forages was not observed on nutrient digestibility, as these parameters linearly decreased with increasing proportions of red clover ( < 0.001). As a result, positive associative effects were observed on daily voluntary intake of digestible DM ( < 0.001). Taken together, the results indicate that the synergy on voluntary intake was due to a greater motivation of animals to eat mixtures (seen in increased intake rates) rather than a positive associative effect on digestive efficiency. Methane yield decreased linearly with increasing proportion of red clover ( < 0.001) and ranged from 20.0 g/kg DMI for sheep fed 100% orchardgrass to 16.1 g/kg DMI for sheep fed 100% red clover. Mixtures of orchardgrass and red clover quadratically ( = 0.03) decreased urinary nitrogen losses and tended ( = 0.099) to quadratically increase nitrogen retention. This synergy between orchardgrass and red clover silages could improve animal performances in addition to the known agronomic benefits of grass-legume mixtures.

Screening of plants from diversified natural grasslands for their potential to combine high digestibility, and low methane and ammonia production.

A total of 156 plant species from 35 botanical families collected from diversified grasslands in the French Massif Central were screened in vitro for their potential to combine high nutritive value for ruminants and a reduced impact on the environment. The vegetative part of plants were analyzed for their chemical composition and incubated in a batch system containing buffered rumen fluid at 39°C for 24 h. The gas production and composition were recorded, and the fermentation end-product concentrations in the incubation medium and the in vitro true organic matter digestibility (IVTOMD) were determined. The results were expressed relative to perennial ryegrass (PRG) values used as a reference. We observed that no relationship between methane (CH4) and volatile fatty acids (VFA) was evidenced for 12 plants, the fermentation of these plants producing significantly less CH4 for a similar level of VFA production. In all, 13 plants showed 50% less CH4 production per unit of organic matter truly digested (OMTD) than PRG. Among these plants, two reduced CH4 by more than 80% and four species had an IVTOMD higher than 80%. The underlying modes of action seem to be different among plants: some result in an accumulation of H2 in the fermentation gas, but others do not. In terms of nitrogen (N) use efficiency, the fermentation of 37 plants halved the ratio between ammonia (N-NH3) and plant N content compared with PRG, of which six showed a complete absence of N-NH3 in the medium. Among these plants, four maintained the IVTDMO at values not significantly different from PRG (P>0.05). Considering the multi-criteria selection, 16 plants showed simultaneously a reduction of more than 80% in N-NH3 production and 30% in CH4 emission per unit of OMTD compared with PRG, including three with an IVTOMD higher than 80%. Overall, the botanical families that reduced simultaneously CH4 and N-NH3 most efficiently were the Rosaceae, Onagraceae, Polygonaceae and Dipsacaceae. The Onagraceae also gave high values for IVTOMD.

Occurrence of associative effects between grasses and legumes in binary mixtures on in vitro rumen fermentation characteristics.

When animals are fed a grass and legume mixture, digestive interactions can occur in the rumen between the substrates contained in the different plants, and the response of the animal to the combination of forages can differ from the balanced median values of their components considered individually. Our objective was to assess the associative effects between temperate forages in 8 grass-legume binary combinations on in vitro rumen fermentation characteristics to highlight synergies or antagonisms in terms of nutritional and environmental impacts. Two grasses (cocksfoot, CF; ryegrass, RG) and 4 legume species (alfalfa, A; white clover, WC; red clover, RC; sainfoin, SAN) were incubated alone and in grass-legume mixture (1:1, wt/wt) in batches containing buffered rumen fluid during 3.5 and 24 h. For each substrate and each incubation time, parameters describing the degradation of the energetic and nitrogenous compounds and their partition into fermentation end products were measured. Data were subjected to ANOVA using a mixed procedure to test quadratic contrasts. At 3.5 h of incubation, many quadratic effects were observed. The presence of A, WC, or RC in mixtures quadratically increased the NH(3)-N production (up to +28% when compared with the calculated value, P < 0.001), whereas the presence of condensed tannin (CT) activity in SAN considerably decreased it (up to -67%, quadratic effect, P < 0.001) and the N disappearance. To a lesser extent, the presence of SAN reduced the proportion of methane in the gas produced (up to 7%, quadratic effect, P = 0.018). Generally, the degradation of OM and NDF was not stimulated by mixing grass and legume, except for SAN. However, the presence of SAN severely impaired the NDF digestion at the early phase of fermentation. At 24 h of incubation, few associative effects were observed in comparison with those observed at 3.5 h of incubation, but the effect of CT of SAN on N metabolism was still clearly present. This study shows that, among the grass-legume mixtures tested in vitro, only the presence of SAN can interact with the grasses to reduce the degradation of proteins and the production of methane but with a transitory negative effect on fiber digestion.

Cell wall component and mycotoxin moieties involved in the binding of fumonisin B1 and B2 by lactic acid bacteria.

AIMS: The ability of lactic acid bacteria (LAB) to bind fumonisins B1 and B2 (FB1, FB2) in fermented foods and feeds and in the gastrointestinal tract could contribute to decrease their bioavailability and toxic effects on farm animals and humans. The aim of this work was to identify the bacterial cell wall component(s) and the functional group(s) of FB involved in the LAB-FB interaction. METHODS AND RESULTS: The effect of physicochemical, enzymatic and genetic treatments of bacteria and the removal/inactivation of the functional groups of FB on toxin binding were evaluated. Treatments affecting the bacterial wall polysaccharides, lipids and proteins increased binding, while those degrading peptidoglycan (PG) partially decreased it. In addition, purified PG from Gram-positive bacteria bound FB in a manner analogue to that of intact LAB. For FB, tricarballylic acid (TCA) chains play a significant role in binding as hydrolysed FB had less affinity for LAB. CONCLUSIONS: Peptidoglycan and TCA are important components of LAB and FB, respectively, involved in the binding interaction. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactic acid bacteria binding efficiency seems related to the peptide moiety structure of the PG. This information can be used to select probiotics with increased FB binding efficiency.

Associative effects between forages on feed intake and digestion in ruminants.

The feeding value of forage mixtures from permanent and temporary multi-species grasslands cannot always be precisely defined. Indeed, the digestibility and feed intake of a combination of forages can differ from the balanced median values calculated from forages considered separately. In order to present an overview of the associative effects between forages on digestion and intake, a literature study was carried out. The associative effects can be studied in a complementary way in vitro to test digestive interactions of a large number of mixtures and to carry out explanatory experiments, and in vivo to investigate intake and digestion at the whole animal scale. We identified three main situations in which interactions between forages can lead to associative effects on intake and digestion: (i) increased intake that can be observed with grass and legume association can be explained by fast digestion of the soluble fraction of legumes, and a higher rate of particle breakdown and passage through the rumen, (ii) increased digestion when a poor forage is supplemented by a high nitrogen content plant can be explained by stimulation of the microbial activity and (iii) modification of digestive processes in the rumen, including proteolysis and methane production when certain bioactive secondary metabolites such as tannins, saponins or polyphenol oxidase are present. According to the type and concentration of these compounds in the diet, the effects can be favourable or unfavourable on intake and digestive parameters. Reported associative effects between forages show a large variability among studies. This reflects the complexity and multiplicity of nutritional situations affecting intake and the rumen function in a given animal. In order to provide more reliable information, further accumulation of data combining in vitro and in vivo studies is required. A better understanding of the associative effects between forages could help to optimise feed use efficiency, resulting in greater productivity, a reduction of the environmental impact of animal emissions and more sustainable animal production.

Screening of fermentative bacteria for their ability to bind and biotransform deoxynivalenol, zearalenone and fumonisins in an in vitro simulated corn silage model.

Fermentative bacteria can potentially be utilized to detoxify corn silage contaminated by Fusarium toxins. The objective of the present study was to test a large number of these bacteria for their ability to bind and/or biotransform deoxynivalenol (DON), zearalenone (ZEN) and fumonisins B(1) and B(2) (FB(1), FB(2)) in conditions simulating corn silage. A total of 202 strains were screened in contaminated, pH 4, corn infusion inoculated with 5 x 10(8) CFU ml(-1). Eight Lactobacilli and three Leuconostoc biotransformed ZEN into alpha-zearalenol, but no biotransformation was detected for DON and fumonisins. In contrast, most strains were capable of binding Fusarium toxins. The most effective genera were Streptococcus and Enterococcus, capable of binding up to 33, 49, 24 and 62% of DON, ZEN, FB(1) and FB(2), respectively. The ability to bind Fusarium toxins seems to be a common property of fermentative bacteria and could help to decrease their toxicity in animals.

Binding of Fusarium mycotoxins by fermentative bacteria in vitro.

AIMS: Fusarium toxins can occur in conserved forages impairing farm animal performances and health. On-farm biological decontamination methods could be an alternative to traditional physico-chemical methods. In this work, the ability to remove Fusarium toxins by fermentative bacteria was evaluated in vitro. METHODS AND RESULTS: Twenty-nine strains of lactic (LAB) and propionic acid bacteria (PAB) were tested for their ability to remove deoxynivalenol (DON) and fumonisins B1 and B2 (FB1, FB2) from an acid, pH 4, medium. Mycotoxin removal was widespread for LAB, but differences among strains were large. Removal was up to 55% for DON, 82% for FB1 and 100% for FB2. Selected strains were also capable of removing up to 88% zearalenone. The PAB strains were less efficient than the LAB. Binding, not biodegradation appeared to be the mode of action, as no toxin derivatives were observed and removal was not impaired in nonviable bacteria. Binding was not affected by pH, except for fumonisins that decreased to nearly 0% at neutral pH. CONCLUSIONS: Selected fermentative bacteria are able to bind main Fusarium mycotoxins. SIGNIFICANCE AND IMPACT OF THE STUDY: The binding ability of selected strains could be used to decrease the bioavailability of toxins in contaminated silages.