Comparison of sheep and dairy cows for in vivo digestibility of perennial ryegrass.

Sheep are often used as a proxy for dairy cows when measuring the digestibility of a feed. In recent years grassland management guidelines for ruminant animals have been re-evaluated in accordance with the progression in animal genetics and the acknowledgement that genetic potential has an influence on both feed intake and digestibility. Recommended pre-grazing herbage mass (HM) targets are now much lower with improved perennial ryegrass varieties available for grazing swards. The objective of this study was to compare the in vivo digestibility of perennial ryegrass in wether sheep and lactating dairy cows. The experimental design was selected to measure the effect of animal species (cows, sheep), sward HM measured cutting herbage at 4 cm above ground level (low: 1 700 kg DM/ha and high: 4 000 kg DM/ha) and season (Spring: Apr-May, Summer: Jul-Aug) on the digestibility of perennial ryegrass. Each HM treatment was offered to each animal within species and season for 12 d using a 2 HM × 2 period changeover Latin square design. There were eight cows and eight sheep, so there were four 2 × 2 Latin squares for each animal species (two) at each season (two), giving 64 observations. During each 12 d experimental period, the first 6 d were used for adaptation (adaptation phase) and the final 6 d were used for measurement (measurement phase). In vivo organic matter digestibility (OMD) in spring did not differ between animal species but in summer sheep had higher in vivo OMD than cows. The results described herein highlight the suitability of wether sheep as an alternative to dairy cows for determining the digestibility of perennial ryegrass in spring but not in summer. Stage of growth of the plant, which is intrinsically linked to season, should be considered as results show that digestibility in the ruminant was affected by season but not differentially affected by changing sward HM.

Digestibility contributes to between-animal variation in feed efficiency in beef cows.

Residual feed intake (RFI) is an alternative measure of feed efficiency (FE) and is calculated as the difference between actual and expected feed intake. The biological mechanisms underlying animal-to-animal variation in FE are not well understood. The aim of this study was to investigate the digestive ability of beef cows selected for RFI divergence as heifers, using two contrasted diets. Fifteen 4-year-old beef cows were selected from a total of 69 heifers based on their RFI following the feedlot test. The selected heifers were ranked into high-RFI (+ 1.02 ± 0.28, n = 8) and low-RFI (-0.73 ± 0.28, n = 7), and a digestibility trial was performed after their first lactation. Both RFI groups were offered two different diets: 100% hay or a fattening diet which consisted of a DM basis of 67% whole-plant maize silage and 33% high starch concentrates over four experimental periods (two per diet). A diet effect was observed on feed intake and apparent digestibility, whereas no diet × RFI interaction was detected (P > 0.05). Intake and apparent digestibility were higher in cows fed the fattening diet than in those fed the hay diet (P < 0.0001). DM intake (DMI) and organic matter apparent digestibility (OMd) were repeatable and positively correlated between the two subsequent periods of measurements. For the hay and fattening diets, the repeatability between periods was r = 0.71 and r = 0.73 for DMI and r = 0.87 and r = 0.48 for OMd, respectively. Moreover, both intake (r = 0.55) and OMd (r = 0.54) were positively correlated (P < 0.05) between the hay and fattening diets. Significant differences between beef cows selected for divergence in RFI as heifers were observed for digestive traits (P < 0.05), DM and organic matter (OM) apparent digestibility being higher for low-RFI cows. Overall, this study showed that apparent digestibility contributes to between-animal variation in FE in beef cows.

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.

Effect of maturity and hybrid on ruminal and intestinal digestion of corn silage in dry cows.

The aim of this study was to evaluate the effect of stage of maturity at harvest on extent of starch, neutral detergent fiber (NDF), and protein digestion, and rumen fermentation in dry cows fed whole-plant corn silage from different hybrids. Four nonlactating Holstein cows cannulated at the rumen and proximal duodenum were fed 4 corn silages differing in hybrid (flint vs. flint-dent) and maturity stage (early vs. late) in a 4 × 4 Latin square design. From early to late maturity, starch content increased (from 234.5 to 348.5 g/kg), whereas total-tract (99.7 to 94.5%) and ruminal starch digestibility (91.3 to 86.5%) decreased significantly. The decrease in ruminal starch digestibility with increasing maturity was similar between hybrids. No effects were found of maturity, hybrid, or maturity × hybrid interaction on total-tract NDF digestibility, ruminal NDF digestibility, true digestibility of N and organic matter in the rumen, or microbial synthesis. Harvesting at later maturity led to increased ruminal ammonia, total volatile fatty acid concentrations, and acetate/propionate ratio but not pH. This study concludes that delaying date of harvest modifies the proportions of digestible starch and NDF supplied to cattle. Adjusting date of corn harvest to modulate amount of rumen-digested starch could be used as a strategy to control nutrient delivery to ruminants.

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.

Food sensory characteristics: their unconsidered roles in the feeding behaviour of domestic ruminants.

When domestic ruminants are faced with food diversity, they can use pre-ingestive information (i.e. food sensory characteristics perceived by the animal before swallowing the food) and post-ingestive information (i.e. digestive and metabolic consequences, experienced by the animal after swallowing the food) to evaluate the food and make decisions to select a suitable diet. The concept of palatability is essential to understand how pre- and post-ingestive information are interrelated. It refers to the hedonic value of the food without any immediate effect of post-ingestive consequences and environmental factors, but with the influence of individual characteristics, such as animal's genetic background, internal state and previous experiences. In the literature, the post-ingestive consequences are commonly considered as the main force that influences feeding behaviour whereas food sensory characteristics are only used as discriminatory agents. This discriminatory role is indeed important for animals to be aware of their feeding environment, and ruminants are able to use their different senses either singly or in combination to discriminate between different foods. However, numerous studies on ruminants' feeding behaviour demonstrate that the role of food sensory characteristics has been underestimated or simplified; they could play at least two other roles. First, some sensory characteristics also possess a hedonic value which influences ruminants' intake, preferences and food learning independently of any immediate post-ingestive consequences. Further, diversity of food sensory characteristics has a hedonic value, as animals prefer an absence of monotony in food sensory characteristics at similar post-ingestive consequences. Second, some of these food sensory characteristics become an indicator of post-ingestive consequences after their initial hedonic value has acquired a positive or a negative value via previous individual food learning or evolutionary processes. These food sensory characteristics thus represent cues that could help ruminants to anticipate the post-ingestive consequences of a food and to improve their learning efficiency, especially in complex environments. This review then suggests that food sensory characteristics could be of importance to provide pleasure to animals, to increase palatability of a food and to help them learn in complex feeding situations which could improve animal welfare and productivity.

Mixing sainfoin and lucerne to improve the feed value of legumes fed to sheep by the effect of condensed tannins.

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).

Effects of condensed tannins in wrapped silage bales of sainfoin (Onobrychis viciifolia) on in vivo and in situ digestion in sheep.

The objective of this study was to characterize the condensed tannins (CTs) in wrapped silage bales of sainfoin (Onobrychis viciifolia) and examine their potential action on in vivo and in situ digestive characteristics in sheep. Silage was made from sainfoin, cut at two phenological stages. The first phenological stage, at which silage was made, was from the first vegetation cycle at the end of flowering and the second stage silage was made from regrowth, 5 weeks after the first cut, but before flowering. The silages made from the two phenological stages were fed to 12 rumen-fistulated sheep in a crossover design. Of the 12 sheep, six received polyethylene glycol (PEG) to bind with and remove the effects of CT, whereas the other six were dosed with water. Organic matter digestibility, total-tract N digestibility and N (N) balance were measured over 6 days. Kinetic studies were performed on total N, ammonia N (NH3-N) and volatile fatty acids (VFAs) in rumen fluid before and 1.5, 3 and 6 h after feeding. The kinetics of degradation of dry matter and N from Dacron bags suspended in the rumen were also determined. Biological activity of CT (protein-binding capacity) and CT concentration were greater for the silage made from sainfoin at the early flowering stage. Total-tract N digestibility was increased by the addition of PEG (P < 0.001) to the sainfoin silage before flowering (P < 0.001). CTs decreased N excretion in urine (P < 0.05) and increased faecal N excretion (P < 0.001), but had no effect on body N retention, which is beneficial for the animal. Ruminal N degradability was smaller in the presence of active CT (P < 0.001) at both phenological stages; however, soluble N (P = 0.2060) and NH3-N (P = 0.5225) concentrations in rumen fluid remained unchanged. The results of this experiment indicate that CT in the sainfoin retain their ability to affect the nutritive value of preserved forage legumes.

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.

NIRS prediction of the feed value of temperate forages: efficacy of four calibration strategies.

Near infrared reflectance spectroscopy (NIRS) of 924 fresh temperate forages were used to develop calibration models for chemical composition - crude ash (CA) and crude protein (CP) - organic matter digestibility (OMD) and voluntary intake (VI). We used 110 samples to assess the models. Four calibration strategies for determining forage quality were compared: (i) species-specific calibration, (ii) family-specific calibration, (iii) a global procedure and (iv) a local approach. Forage calibration data sets displayed CA values ranging from 52 to 205 g/kg of dry matter (DM), CP values from 50 to 280 g/kg DM, OMD values from 0.48 to 0.85 g/g and VI values from 22.5 to 115.2 g DM/kg metabolic body weight (BW0.75). The calibration models performed well for all the variables except for VI. For CA, local procedure showed lower standard error of prediction (SEP) than species-specific, family-specific or global models. For CP, the calibration models all showed similar SEP values (11.13, 11.08, 11.38 and 11.34 g/kg DM for species-specific, family-specific, global and local approaches). For OMD, the local procedure gave a similar SEP (0.024 g/g) to specific species and global procedures (0.027 g/g) and a lower SEP than the family-specific approach (0.028 g/g). For VI, the local approach and species-specific calibration showed lower SEP (7.08 and 7.16 g/kg BW0.75) than the broad-based calibrations (8.09 and 8.34 g/kg BW0.75 for family-specific model and global procedure, respectively). Local calibration may thus offer a practical way to develop robust universal equations for animal response determinations.

Sheep use preingestive cues as indicators of postingestive consequences to improve food learning.

Previous work has shown that herbivores successfully learn to associate food sensory characteristics with postingestive consequences when the conditioning procedure is simple, whereas this ability breaks down when the learning task is made more complex. We hypothesized that sensory characteristics could act as indicators of postingestive consequences and that the presence of preingestive cues would improve the food learning of sheep in situations varying in complexity. Sixteen sheep were subjected to a first conditioning phase to associate 2 flavors added to alfalfa hay with either a positive or a negative consequence, induced by intraruminal administration of starch (330 mg/g of DMI) or LiCl (5 mg/g of DMI). Sheep progressively decreased their choice of the flavored hay associated with the negative consequence (P < 0.05). This procedure provided sheep with experience with postingestive consequences associated with the different flavors. In a second conditioning phase, the experienced sheep and 16 naïve sheep were divided into groups of 8 and subjected to either a simple or a complex conditioning procedure [i.e., the 2 flavors were offered on separate days (simple conditioning) or simultaneously within a day (complex conditioning)]. The 2 flavors applied to grass hay were associated with either positive (starch, 330 mg/g of DMI) or negative (LiCl, 10 mg/g of DMI) consequences. As hypothesized, sheep in the simple conditioning group expressed a greater aversion to the flavored hay associated with the negative consequence than did those in the complex conditioning group (0.303 +/- 0.035 vs. 0.474 +/- 0.035 respectively; P < 0.01). Experienced sheep rejected the flavor associated with the negative consequence more strongly than did naïve sheep, regardless of the conditioning procedure (0.304 +/- 0.029 vs. 0.470 +/- 0.041 respectively; P < 0.05). The initial increased preference for aniseed (0.80 +/- 0.04), however, greatly influenced food learning because sheep negatively conditioned on this flavor expressed less avoidance than those negatively conditioned on orange (0.53 +/- 0.04 vs. 0.25 +/- 0.03 respectively; P < 0.01). In conclusion, the simultaneous scenario was actually perceived as complex by all sheep, and experienced sheep were more efficient in food learning than naïve sheep in both the simple and complex learning contexts. The sheep were thus able to generalize the association between sensory cues and postingestive consequences, especially in a complex environment, and then to use these sensory cues as indicators of postingestive consequences.

Feeding behaviour of sheep fed lucerne v. grass hays with controlled post-ingestive consequences.

Understanding what determines feeding behaviour in herbivores is essential to optimise the use of forages in breeding systems. Herbivores can evaluate foods by associative learning of their pre-ingestive characteristics (taste, odour, etc.) and their post-ingestive consequences. Post-ingestive consequences are acknowledged as influencing intake and food choices, but the role of pre-ingestive characteristics is still being debated. Our experiment was designed to test their separate effects on daily dry matter intake (DMI), intake patterns and short-term choices in sheep by crossing the nature of the hay orally consumed (o) ad libitum, lucerne (L) or grass (G), with the nature of the hay introduced into the rumen (r), L or G, at a rate of half the total amount of hay received the day before. We applied four treatments, Go/Gr, Go/Lr, Lo/Gr and Lo/Lr, to test the effects of (i) post-ingestive consequences with similar pre-ingestive characteristics (Go/Gr v. Go/Lr; Lo/Gr v. Lo/Lr) and (ii) pre-ingestive characteristics with similar post-ingestive consequences at the end of the feeding period (Go/Lr v. Lo/Gr). Six rumen-fistulated sheep underwent all the treatments over 11-day periods in a latin square design. Eating time was restricted to 6 h/day, intraruminal introductions were performed just before food offer and choice tests were conducted after food removal. For similar pre-ingestive characteristics, DMI increased when L hay was introduced into the rumen rather than G (P < 0.05), possibly owing to a lower fill effect of L due to its lower NDF content and higher rumen degradability. The increased DMI resulted from longer eating time when G was orally consumed (149 v. 192 min, P < 0.05), whereas it resulted from higher intake rate with L (4.8 v. 6.1 g/min, P < 0.05). For similar post-ingestive consequences at the end of the feeding period (Go/Lr and Lo/Gr), DMI were similar (P > 0.05). Pre-ingestive characteristics or palatability per se did not therefore influence daily intake, although they influenced eating patterns. Pre-ingestive characteristics also greatly influenced short-term choices in favour of the hay that was not previously consumed, independently of any post-ingestive influence. This study confirms the effects of post-ingestive consequences on daily intake, but demonstrates that these variations are obtained by different behavioural adjustments under the influence of pre-ingestive characteristics. Preference for novelty, regardless of post-ingestive consequences, thus suggests that sheep may seek a diverse diet more for pleasure than for functional purposes, with implications for animal welfare.

Risk of subacute ruminal acidosis in sheep with separate access to forage and concentrate.

This study aimed to investigate whether sheep offered free-choice intake of forage and concentrate develop subacute ruminal acidosis (SARA) and to identify SARA-associated feeding behavior components. In a crossover design over two 28-d periods, 11 rumen-cannulated wethers received wheat and alfalfa hay in 2 separate compartments. Concentrate and forage were provided for ad libitum access or in a fixed amount corresponding to 80% of ad libitum hay intake with a concentrate:forage ratio of 60:40 on a DM basis. In both diets, sheep were fed 2 equal portions at 0800 and 1600 h. Ruminal pH, voluntary intake, and feeding behavior were recorded continuously from d 1 to 9 and d 15 to 23 in each period. When no measurements were performed, the animals were housed in larger pens with straw bedding. When fed for ad libitum intake, the sheep ingested 1,340 g of DM/d consisting of 49.1% wheat, whereas with the fixed diet they ate 872 g of DM/d consisting of 58.4% wheat. Sheep fed for ad libitum intake spent more time with ruminal pH < 5.6 than when fed in fixed amounts (7.77 vs. 3.05 h/d, P < 0.001). The time spent with ruminal pH <5.6 was mainly linked to the amount of feed ingested and especially the amount of wheat (P < 0.001). Our results suggest that when fed for ad libitum intake with free choice wheat, the achieved concentrate:forage ratio of near 50:50 and a larger hay intake enable sheep to consume more wheat. When sheep were fed for ad libitum intake, feeding bouts were spread evenly throughout the day. Although ruminal pH reached the same minimum level in both diets after main meals, time to reach pH nadir was longer with ad libitum diet (P < 0.001). In addition, after reaching this minimum value, ruminal pH increased more slowly in this diet, inducing a decreased preprandial ruminal pH (P < 0.001). Consequently, the ad libitum diet led to a longer time below pH 5.6. A slow decrease in ruminal pH may enable sheep to consume larger quantities of food. However, free access to concentrate maintains continuously elevated content of ruminal fermentation end products and so requires more time for pH to return to neutral values. Thus, interval between feed distributions should be as large as possible to help resume the preprandial ruminal pH and to limit time spent with pH <5.6.

Diet learning through post-ingestive consequences in sheep: the case of starch and casein variously combined in the same foods.

Ruminants generally succeed in selecting an appropriate diet relative to their nutrient requirements, and macronutrients such as energy and protein are considered as playing a primary role in their food preferences. Diet learning through post-ingestive consequences, evidenced by many studies that involved situations with simple discrimination tasks, may explain this ability to select a suitable diet. However, in more complex feeding environments where animals are faced with many plants varying both in their energy and protein contents, the extent to which such a mechanism operates is still uncertain. Our objective was to investigate a more complex feeding situation than previously tested and to determine whether lambs were able to associate three forages with both energy-based and protein-based post-ingestive consequences of various intensities. The doses of starch and casein used to elicit these post-ingestive consequences were chosen so that the apparent metabolisable energy and crude protein values of hays were within the range found in plants normally encountered under field conditions. We applied five treatments: EmPm, EhPm, EmPh, EhPh, where medium (m) or high (h) levels of nutrients (E = energy and P = protein) were associated with different hays, and E0N0 where no stimuli were associated. Each animal experienced only three treatments: E0N0-EmPm-EhPm or E0N0-EmPh-EhPh, each one being associated with one hay during conditioning, on the basis of one treatment per day over three consecutive days. Animals then had a choice between the hay associated with E0N0 and one of the two others, successively on the two following days. This procedure was repeated through five 1-week-long periods, and applied to two groups of lambs (n = 12) maintained at feeding levels sufficient for maintenance or for a 150 g/day growth rate. This experiment was complemented by a methodological trial that aimed to estimate the doses of starch and casein eliciting preferences and a post-trial that aimed to evaluate the effect of our treatments on sheep ruminal environment. The methodological trial showed that lambs perceived the stimuli and the doses, and the post-trial showed that treatments affected the ruminal environment. In the main experiment, the lambs preferred the rewarded hays but did not discriminate in their choices the variation of post-ingestive consequences due to energy and protein. This suggests that in situations closer to reality where animals experience many foods varying in nutrient contents, the learning process may be impaired. The lambs do not seem to have strictly relied on it for shaping their diet choices.

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.

Voluntary intake and digestibility in horses: effect of forage quality with emphasis on individual variability.

Food intake is a key biological process in animals, as it determines the energy and nutrients available for the physiological and behavioural processes. In herbivores, the abundance, structure and quality of plant resources are known to influence intake strongly. In ruminants, as the forage quality declines, digestibility and total intake decline. Equids are believed to be adapted to consume high-fibre low-quality forages. As hindgut fermenters, it has been suggested that their response to a reduction in food quality is to increase intake to maintain rates of energy and nutrient absorption. All reviews of horse nutrition show that digestibility declines with forage quality; for intake, however, most studies have found no significant relationship with forage quality, and it has even been suggested that horses may eat less with declining forage quality similarly to ruminants. A weakness of these reviews is to combine data from different studies in meta-analyses without allowing the differences between animals and diets to be controlled for. In this study, we analysed a set of 45 trials where intake and digestibility were measured in 21 saddle horses. The dataset was analysed both at the group (to allow comparisons with the literature) and at the individual levels (to control for individual variability). As expected, dry matter digestibility declined with forage quality in both analyses. Intake declined slightly with increasing fibre contents at the group level, and there were no effects of crude protein or dry matter digestibility on intake. Overall, the analysis for individual horses showed a different pattern: intake increased as digestibility and crude protein declined, and increased with increasing fibre. Our analysis at the group level confirms previous reviews and shows that forage quality explains little of the variance in food intake in horses. For the first time, using mixed models, we show that the variable 'individual' clarifies the picture, as the horses showed different responses to a decrease in forage quality: some compensated for the low nutritional value of the forages by increasing intake, few others responded by decreasing intake with declining forage quality, but not enough to cause any deficit in their energy and protein supplies. On the whole, all the animals managed to meet their maintenance requirements. The individual variability may be a by-product of artificial selection for performance in competition in saddle horses.

Sensitivity of galanin- and melanin-concentrating hormone-containing neurones to nutritional status: an immunohistochemical study in the ovariectomized ewe.

The sensitivities of galanin and melanin-concentrating hormone (MCH) neuronal systems to nutrition are poorly understood in sheep compared to rodents. The aim of this study was to describe the changes in the numbers of galanin and MCH neurones in ovariectomized ewes submitted to different nutritional levels. In the first experiment, ewes were fed ad libitum or food deprived for 24 h. In the second experiment, two groups of ewes were fed at maintenance level (group 100) or undernourished (group 40) for 167 days, after which one-half of each group was killed or refed ad libitum (group 100R and 40R) for 4 days. The MCH neuronal population located in the lateral hypothalamic area was not affected by these nutritional changes. Long-term undernutrition enhanced the number of galanin neurones located in the infundibular nucleus and the dorsal hypothalamic area (DHA), refeeding resulted in an increase of neurones in the DHA and preoptic area, but short-term starvation had no effect on any galanin subpopulations. Our data suggest that the sensitivity of MCH neuronal populations to nutrition in sheep differs from that of rodents. Various populations of galanin-containing neurones differ in sensitivity in ewes subjected to long undernutrition and refeeding but not to short starvation.

Long-term undernutrition followed by short-term refeeding effects on the corticotropin-releasing hormone containing neurones in the paraventricular nucleus: an immunohistochemical study in sheep.

The effect of nutritional level on the immunoreactivity of corticotropin-releasing hormone (CRH) in neurones of the hypothalamic paraventricular nucleus was described in sheep, a ruminant, whose feeding strategy differs from that of monogastric species. Two groups of ewes were underfed (40%), or fed at maintenance (100%) for 167 days, after which one-half of each group was killed or ad libitum refed (at least 150% of maintenance) for 4 days before killing. The presence of CRH in the paraventricular nucleus was examined by immunohistochemistry. The number of CRH immunoreactive neurones was increased in underfed ewes, but without modification of the plasma concentration of cortisol, indicating that the rise of CRH was not released in the portal blood nor linked to the pituitary-adrenal axis. Refeeding did not modify significantly the number of CRH immunoreactive neurones in the nucleus although these neurones were increased, only in refed ewes that were previously underfed. These data differ from those for rats and mice where CRH expression is decreased or not modified by underfeeding which could reflect different effects of undernutrition on CRH immunoreactive neurones in monogastric compared to ruminants species.