Benefits of barley straw as a forage for dairy calves before and after weaning.

The aim of this study was to assess the potential consequences on calf intake, performance, behavior, ruminal microbiome, and ruminal epithelium development of combining the inclusion of chopped barley straw and alfalfa hay during the pre- and postweaning periods keeping concentrate to forage ratio constant among dietary treatments. Forty-five Holstein calves (44 ± 5.7 kg of body weight [BW] and 3 ± 1.5 d of age) individually penned were blocked by BW and randomly assigned to a common pellet concentrate fed ad libitum along with one of following forage feeding strategies: barley straw before and after weaning (S-S), barley straw before and alfalfa hay after weaning (S-A), or alfalfa hay before and after weaning (A-A). All calves received the same milk replacer regimen. Forage was supplied in a separated bucket at the rate of 7.5% (preweaning) and 15% (postweaning) of total solid feed intake of the previous day. Feed intake and BW were recorded daily and weekly, respectively. Rumen samples were obtained via a stomach tube at 53, 66, and 87 d and were composite in 3 samples of 5 animals each for subsequent rumen microbiome analysis. A rumen epithelium sample was taken by endoscopy at 90 d to assess gene expression of OCLN, CLDN4, SLC9A1, SLC9A3, SLC16A1, SLC16A4, IL6, and TGFB1. Data were analyzed with a mixed-effects model accounting for the fixed effects of block, forage, week of study, and their interaction, and calf as a random effect. The type of forage fed did not affect concentrate feed, forage, or total DM intake before weaning. However, S-A and A-A calves consumed less concentrate feed and S-A calves grew at a lower rate after weaning than S-S calves. Expression of the gene coding for SLC16A1 in the rumen epithelium was greatest in S-S among treatments. Rumen microbiome did not differ among treatments, while the relative abundance of Acidaminococcus and Selenomas genera increased, while Alloprevotella, Bifidobaterium, Olsenella, and Succiclasticum genera decreased with age. In conclusion, feeding barley straw before and after weaning was more effective than feeding alfalfa hay in promoting concentrate feed intake after weaning and fostering an increase in the expression of SLC16A1 in the rumen epithelium.

Including 8 hours of access to alfalfa in 1 or 2 grazing sessions in dairy cows fed a partial mixed ration: Effects on intake, behavior, digestion, and milk production and composition.

The aim of this study was to evaluate the inclusion of alfalfa grazing during 8 h continuous or partitioned in 2 separated sessions of 4 h after each milking, on nutrient intake, digestibility, ruminal fermentation, feeding behavior, milk production, milk composition, and milk fatty acid profile, in late-lactation cows fed a partial mixed ration (PMR). Twelve dairy cows (193 ± 83 d in milk, 584 ± 71 kg of body weight) were housed in individual outdoor pens and assigned to treatments according to a 3 × 3 Latin square design replicated 4 times. The treatments were as follows: (1) control (T0), cows were fed a total mixed ration (TMR) provided ad libitum 20.0% crude protein (CP), 32.2% neutral detergent fiber (NDF); (2) fed a diet combining a PMR which had the same ingredient composition as the TMR (60% of ad libitum intake) + 1 session of 8 h of pasture (T8), continuous grazing alfalfa (Medicago sativa; 20.6% CP, 35.8% NDF) after the p.m. milking; and (3) PMR (60% of ad libitum intake) + 2 daily sessions of 4 h of access to pasture after each milking (T4+4). The experiment lasted 57 d and was divided into 3 periods of 19 d each. The first 12 d of each period was used for diet adaptation, and the last 7 d was used for data collection. No differences among treatments were observed for any of the productive variables, feeding efficiency, or purine derivatives excretion. Cows in T0 had greater intake and apparent digestibility of dry matter, organic matter, and nonfibrous carbohydrates compared with T4+4 and T8. Compared with T0, alfalfa grazing increased the concentration of C18:1 trans-11 and decreased those of C16:0 and C17:0 in milk fat. Cows in T4+4 consumed 1.1 more kg DM/d of alfalfa and N provided by alfalfa in the diet was 3 percentage points higher compared with T8 cows (266 vs. 229 g/d, respectively). In addition, T4+4 cows had a greater daily range of ruminal pH than T8 (0.73 vs. 0.93), and the highest concentrations of NH3-N were recorded during the a.m. grazing session while in T8 cows it occurred during the night. In conclusion, including 8 h of alfalfa grazing in T8 and T4+4 treatments allowed the substitution between 35.8 and 38.7% of the total dry matter intake (DMI) of a PMR (with a similar CP concentration to alfalfa) for pasture, maintaining milk solids production and increasing the C18:1 trans-11 of milk fat compared with a TMR in mid late-lactation cows. In an herbage plus PMR diet, splitting the 1 continuous grazing session of 8 h into 2 sessions of 4 h increased the proportion of energy and N provided by alfalfa pasture and reduced PMR intake, without modifying the total nutrient intake or productive performance of cows.

Effects of feed withdrawal duration on animal behaviour, rumen microbiota and blood chemistry in feedlot cattle: implications for rumen acidosis.

Feed withdrawal (FW) is a frequent issue in open outdoor feedlot systems, where unexpected circumstances can limit the animals' access to food. The relationship among fasting period, animal behaviour during feed reintroduction (FR) and acidosis occurrence has not been completely elucidated. Twenty steers fitted with rumen catheters were fed a high-concentrate diet (concentrate : forage ratio 85 : 15) and were challenged by a protocol of FW followed by FR. The animals were randomly assigned to one of the four treatments: FW for 12 h (T12), 24 h (T24), 36 h (T36) or no FW (control group) followed by FR. The steers' behaviour, ruminal chemistry, structure of the ruminal microbial community, blood enzymes and metabolites and ruminal acidosis status were assessed. Animal behaviour was affected by the FW-FR challenge ( P < 0.05). Steers from the T12, T24 and T36 treatments showed a higher ingestion rate and a lower frequency of rumination. Although all animals were suspected to have sub-acute ruminal acidosis (SARA) prior to treatment, a severe case of transient SARA arose after FR in the T12, T24 and T36 groups. The ruminal pH remained below the threshold adopted for SARA diagnosis ( pH value = 5.6) for more than three consecutive hours (24, 7 and 19 h in the T12, T24 and T36 treatments, respectively). The FW-FR challenge did not induce clinical acute ruminal acidosis even though steers from the T36 treatment presented ruminal pH values that were consistent with this metabolic disorder (pH threshold for acute acidosis = 5.2). Total mixed ration reintroduction after the withdrawal period reactivated ruminal fermentation as reflected by changes in the fermentation end-products. Ruminal lactic acid accumulation in steers from the T24 and T36 treatments probably led to the reduction of pH in these groups. Both the FW and the FR phases may have altered the structure of the ruminal microbiota community. Whereas fibrolytic bacterial groups decreased relative abundance in the restricted animals, both lactic acid producer and utiliser bacterial groups increased ( P < 0.05). The results demonstrated a synchronisation between Streptococcus (lactate producer) and Megasphaera (lactate utiliser), as the relative abundance of both groups increased, suggesting that bacterial resilience may be central for preventing the onset of metabolic disturbances such as ruminal acidosis. A long-FW period (36 h) produced rumen pH reductions well below and lactic acid concentration increased well above the accepted thresholds for acute acidosis without any perceptible clinical signs.

Reducing milking frequency in early lactation improved the energy status but reduced milk yield during the whole lactation of primiparous Holstein cows consuming a total mixed ration and pasture.

To investigate the immediate and long-term performance effects of milking frequency during early lactation of primiparous dairy cows consuming a total mixed ration and pasture, 20 Holstein cows were assigned in a randomized block design to either once-daily (1×) or twice-daily (2×) milking during the first 8 wk of lactation (treatment period). After the treatment period, all cows were milked 2× until wk 43 of lactation. Cows were fed a total mixed ration (approximately 15 kg of DM/cow per day) and allowed to graze an oat pasture (Avena sativa). Dry matter intake was 19.1 kg of DM/cow per day on average and was not affected by treatments. Milk yield was 40% lower in cows milked 1× during the treatment period, and a carryover effect existed until wk 21 of lactation, resulting in a final reduction of 15% of milk yield in the whole lactation. Milk lactose concentration decreased, whereas fat and protein concentrations increased for cows milked 1×. Mobilization of energy reserves during the treatment period occurred in both groups, but cows milked 1× showed greater body condition score and greater backfat thickness. In conclusion, milking 1× during the first 8 wk of lactation resulted in immediate and carryover negative effects on milk and milk solid yield without affecting feed intake, resulting in the improved energy status of primiparous dairy cows.

Productive performance and digestive response of dairy cows fed different diets combining a total mixed ration and fresh forage.

The purpose of this experiment was to determine the effects of feeding increasing levels of fresh forage (FF) as a proportion of total dry matter intake (DMI) on nutrient intake, rumen digestion, nutrient utilization, and productive performance of total mixed ration (TMR)-fed cows. Twelve dairy cows (90 ± 22 d in milk, 523 ± 88 kg of body weight, 7,908 ± 719 kg of milk production in the previous lactation) were housed in individual tiestalls and assigned to treatments according to a 3 × 3 Latin square design replicated 4 times. Treatments were 100% TMR (T100), 75% TMR plus 25% FF (T75), and 50% TMR plus 50% FF (T50). The experiment lasted 60 d, divided into 3 periods of 20 d each; the first 12 d of each period were used for diet adaptation and the last 8 d for data collection. The TMR (18.1% crude protein, 24.6% acid detergent fiber) and FF (Lolium multiflorum; 15.1% crude protein, 24.1% acid detergent fiber) were prepared and cut daily and offered to each cow individually. The highest DMI was reached in T100 and T75, which was reflected in greater intake of the different nutrients than T50. No differences were detected in the apparent total digestibility of the nutrients, mean ruminal pH, and total volatile fatty acid concentrations among treatments. Cows in T50 resulted in the lowest ruminal N-NH3 concentration and the lowest microbial N flow to the duodenum. Milk yield was 8.5% higher from cows in T100 and T75 compared with T50, but we observed no differences for milk fat or milk protein yield among treatments. Milk fat of cows fed T50 had 8% more unsaturated fatty acids (FA) than that of cows fed T100, mostly because of a higher content of monounsaturated FA. Additionally, cows in T50 had a higher concentration of linoleic acid, vaccenic acid, and rumenic acid than T100. Meanwhile, the concentration of linoleic acid and vaccenic acid in cows fed T75 was higher than T100. The milk fat of the cows fed T50 and T75 had a lower n-6:n-3 ratio than T100. We concluded that including up to 29% of FF in the total DMI in combination with a TMR did not affect the intake or digestion of nutrients or the productive response in dairy cows and resulted in a higher concentration of desirable FA from a consumer's perspective.

Supplementing high-quality fresh forage to growing lambs fed a total mixed ration diet led to higher intake without altering nutrient utilization.

The effect of supplementing high-quality fresh forage, mainly based on alfalfa, to growing lambs fed with decreasing levels of total mixed ration (TMR) was studied on intake, digestion and ruminal environment. In total, 24 catheterized lambs (25.2±3.67 kg) housed in individual metabolism cages were assigned to one of four treatment diets: 'TMR100': TMR offered ad libitum; 'TMR75' and 'TMR50': TMR at a level of 0.75 and 0.50 of potential intake, respectively, complemented with fresh forage without restriction; 'TMR0': only fresh forage ad libitum. The feeding behavior, nutrient intake and digestibility, kinetics of passage and rumen environment were evaluated. As the level of TMR in the diet decreased, lambs increased the forage intake and spent more time eating and ruminating, less time resting and demonstrated a higher rate of intake. Those changes resulted in a higher nutrient intake of dry matter, organic matter, nitrogen, NDF and ADF, but a slightly lower organic matter digestibility, while no differences were detected in the output rate of particles. As a consequence, with the decrease of TMR and increase of forage intake, the ingested energy increased. Higher ruminal pH and NH3-N concentrations were observed for lower levels of TMR in the diet. The total volatile fatty acids, acetate and propionate concentrations presented a quadratic response. Total volatile fatty acids and acetate concentrations were higher and propionate concentration was lower in lambs consuming mixed diets (TMR50 and TMR75). We concluded that the inclusion of high-quality fresh forage in a combined diet with TMR in lambs had positive effects on nutrient intake without negative consequences on digestion and rumen environment.

Combination of legume-based herbage and total mixed ration (TMR) maintains intake and nutrient utilization of TMR and improves nitrogen utilization of herbage in heifers.

Diets combining herbage and total mixed rations (TMR) are increasingly used in temperate regions for feeding ruminants, but little information is available regarding the effects on nutrient intake and digestion of this feeding management in beef cattle. The aim of this study was to determine the effects of combining TMR (10% CP and 13% ADF), and legume-based herbage (14% CP and 27% ADF) on intake, nutrient digestion, ruminal fermentation, microbial N flow and glucose and nitrogen metabolism in heifers. The experiment was a 3×3 Latin square design replicated three times; each period lasted 18 days (10 adaptation days and 8 measurement days). Nine cross-bred (Aberdeen Angus×Hereford) heifers (214±18 kg) fitted with permanent rumen catheters and housed in individual metabolic cages were assigned to one of three treatments: 24 h access to TMR ( T ), 24 h access to herbage ( H ) or combined diets with 18 h access to TMR and 6 h access to herbage ( T+H ). Data were evaluated using a mixed model. Animals fed T+H (TMR 71% and herbage 29%) diets tended to have a higher dry matter intake as a proportion of their BW than animals fed T. The T+H diet did not change ruminal fermentation (pH, N-NH3 and volatile fatty acids) or the N metabolism relative to the T diet, but increased the glucagon concentration and altered glucose metabolism. Conversely, animals fed T+H had increased purine derivatives excretion, increased N use efficiency for microbial protein synthesis and decreased plasma urea and urinary N excretion relative to animals fed H diet. The use of combined diets led to consumption of nutrients similar to a TMR diet, without reducing nutrient use and could improve N utilization compared with the herbage-only diet.

Digestive response of dairy cows fed diets combining fresh forage with a total mixed ration.

The objective of this experiment was to quantify the response of dairy cows fed a total mixed ration (TMR) to increasing access to high-quality temperate fresh forage with respect to energy intake, rumen fermentation, microbial protein flow, passage rate, nutrient digestion and utilization, and metabolic and endocrine profiles. Nine Holstein cows fed a TMR were assigned to the following treatments according to a 3×3 Latin square replicated 3 times with 20-d periods and sampling on the last 10 d of each period: 0 (T0), 4 (T4), or 8 (T8) h of daily access to fresh forage. The forage (Lolium multiflorum; 17.1% crude protein, 26.5% acid detergent fiber) was cut daily and offered ad libitum beginning at 0800h, and a TMR (16.1% crude protein, 22.9% acid detergent fiber) was offered ad libitum during the remaining time. Energy intake and balance were higher in T0 than in T8, which was reflected in higher blood glucose and insulin concentrations in T0. Total volatile fatty acid concentrations in the rumen were higher in T0 and T4 than in T8, pH was lower in T4 than in T8, and ammonia-N was higher in T0 than in T8. No differences among treatments were detected in microbial protein flow to the duodenum, digestibility of nutrients, apparent efficiency of energy, or N utilization for milk production, but the total mean retention time of feed in the digestive tract was higher in T8 than in T0. It is concluded that more than 4h of daily access to high-quality fresh forage in the diet of dairy cows fed a TMR reduced energy intake and balance but had no effects on nutrient digestion or utilization.

Short communication: Intake, milk production, and milk fatty acid profile of dairy cows fed diets combining fresh forage with a total mixed ration.

To establish the effects of access time to high-quality temperate fresh forage (FF) on intake and performance of cows fed a total mixed ration (TMR), 9 Holstein cows were assigned to three 3 × 3 Latin squares with 20-d periods with sampling in the last 10 d. The following treatments were evaluated: 0 (T0), 4 (T4), and 8 (T8) h of daily access to FF. Forage (Lolium multiflorum) was cut daily and offered ad libitum beginning at 0800 h in individual stalls, and a TMR was offered ad libitum during the period when cows had no access to FF. Fresh forage dry matter intake (DMI) increased and TMR DMI decreased with the access time to FF, whereas total DMI was 3.0 kg higher for T4 than for T8; no differences were detected with T0. Compared with T0, 8h/d of access to FF decreased milk, protein, and casein yields, and tended to decrease fat and lactose yields, but treatment had no effect on milk composition or feed efficiency. The milk fatty acid profile in T8 had a higher content of rumenic, vaccenic, and linolenic acids compared with T0. We concluded that cows with 4h of access to high-quality FF had similar DMI and milk yields as cows fed only the TMR, but more than 4h of access reduced DMI and performance without changes in feed efficiency, although milk fat had enhanced levels of beneficial fatty acids.

Effect of sorghum grain supplementation on glucose metabolism in cattle and sheep fed temperate pasture.

The aim of this work was to evaluate the effect of sorghum grain supplementation on plasma glucose, insulin and glucagon concentrations, and hepatic mRNA concentrations of insulin receptor (INSR), pyruvate carboxylase (PC), and phosphoenolpyruvate carboxykinase (PCK1) mRNA and their association with nutrient intake, digestion and rumen volatile fatty acids (VFA) in cattle and sheep fed a fresh temperate pasture. Twelve Hereford × Aberdeen Angus heifers and 12 Corriedale × Milchschaf wethers in positive energy balance were assigned within each species to one of two treatments (n = 6 per treatment within specie): non-supplemented or supplemented with sorghum grain at 15 g/kg of their body weight (BW). Supplemented cattle had greater plasma glucose concentrations, decreased plasma glucagon concentrations and tended to have greater plasma insulin and insulin-to-glucagon ratio than non-supplemented ones. Hepatic expression of INSR and PC mRNA did not differ between treatments but PCK1 mRNA was less in supplemented than non-supplemented cattle. Supplemented sheep tended to have greater plasma glucagon concentrations than non-supplemented ones. Plasma glucose, insulin, insulin-to-glucagon ratio, and hepatic expression of INSR and PC mRNA did not differ between treatments, but PCK1 mRNA was less in supplemented than non-supplemented sheep. The inclusion of sorghum grain in the diet decreased PCK1 mRNA but did not affect PC mRNA in both species; these effects were associated with changes in glucose and endocrine profiles in cattle but not in sheep. Results would suggest that sorghum grain supplementation of animals in positive energy balance (cattle and sheep) fed a fresh temperate pasture would modify hepatic metabolism to prioritize the use of propionate as a gluconeogenic precursor.

Use of NaHCO3 and MgO as additives for sheep fed only pasture for a restricted period of time per day: effects on intake, digestion and the rumen environment.

Effects of NaHCO3 and MgO buffer addition on intake and digestive utilization of a pasture were studied in wethers allowed a restricted time of access to forage. Twelve wethers housed in metabolic cages and fed fresh forage (predominantly Lotus corniculatus) ad libitum for 6 h/d were randomly assigned to one of the following treatments: a control forage without buffer (C) or a forage plus buffer composed of a mixture of 750 g/kg NaHCO3 and 250 g/kg MgO at 20 g/kg dry matter intake (B). Feeding behaviour, feed and water intake and digestibility, urine output, Na urine elimination, kinetics of passage, ruminal pH and ammonia concentration, N balance and ruminal microbial N synthesis were determined in vivo, and the ruminal liquor activity was evaluated in vitro by fermentation of wheat straw. Addition of buffer increased total water intake (p = 0.05), Na urinary output (p = 0.01), purine derivative excretion in urine (p = 0.05) and tended to decrease mean total retention time in the digestive tract (p = 0.09). However, buffer addition increased ruminal pH (p < 0.001) and tended to decrease the ammonia concentration (p = 0.05). That use of buffer decreased ruminal activity was evidenced by a lower volume of gas produced in vitro (p = 0.01) possibly due to a lower microbial concentration in rumen liquor. The higher rumen dilution rate, likely due to a higher water intake, seems to have been the key driver of the actions of buffer supplementation on the rumen environment. Moreover, addition of NaHCO3 led to an increased urinary Na excretion, which should be considered due to its likely negative environmental impacts.

Suitability of live yeast addition to alleviate the adverse effects due to the restriction of the time of access to feed in sheep fed only pasture.

The effect of yeast addition on intake and digestive utilization of pasture was studied in ovines under restricted time of access to forage. Eighteen wethers housed in metabolic cages and fed fresh forage (predominantly Lotus corniculatus) were randomly assigned to three treatments: forage available all day (AD); forage available only 6 h/day (R) and forage available only 6 h/day plus live Saccharomyces cerevisiae yeast (RY). Feed intake and digestibility, feeding behaviour, kinetics of passage, ruminal pH and ammonia concentration, nitrogen balance and microbial nitrogen synthesis (MNS) were determined in vivo, and ruminal liquor activity of animals was evaluated in vitro. Restricted animals consumed less than those fed all day but achieved more than 75% of the intake and spent less time ruminating (p = 0.014). Although animals without restriction consumed more feed, they had a lower rate of passage (p = 0.030). The addition of yeast did affect neither intake nor feeding behaviour, but increased digestibility. Organic matter digestibility tended to increase 11% by yeast addition (p = 0.051), mainly by a rise in NDF (27%, p = 0.032) and ADF digestibility (37%, p = 0.051). Ingested and retained N was lower in restricted animals, as MNS (p ≤ 0.045). The use of yeasts did not significantly change the N balance or MNS, but retained N tended to be higher in supplemented animals (p = 0.090). Neither ruminal pH nor ammonia concentrations were affected by the restriction, but restricted animals had a lower ruminal activity evidenced by a lower volume of gas (p = 0.020). The addition of yeast overcame this limitation, noted by a higher volume of gas of inocula from supplemented animals (p = 0.015). Yeast addition emerged as a useful tool to improve digestibility of forage cell walls in ovines under restricted time of access to forage.

Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values.

To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The efficiency of microbial protein synthesis was greater in P2 (P = 0.007). In conclusion, addition of feeds containing NFCs to fresh temperate forage reduced the digestibility of cell walls and did not improve microbial protein synthesis or its efficiency. An increase in these parameters was associated to the early phenological stage of the forage.

Short-term nutritional supplementation of ewes in low body condition affects follicle development due to an increase in glucose and metabolic hormones.

This study tested whether the effects of a short period of nutritional supplementation given to ewes during the luteal phase on follicle development and ovulation rate is associated with an increase in circulating concentrations of FSH, glucose or metabolic hormones. Oestrus was synchronised with two prostaglandin injections given 9 days apart and the supplement consisted of corn grain and soybean meal. Corriedale ewes with low body condition were randomly assigned to 2 groups: the control group (C; n = 10) received a maintenance diet while the short-term supplemented group (STS; n = 10) received double the maintenance diet over days 9 to 14 of the oestrous cycle (day 0 = ovulation). Ovaries were examined daily by ultrasound and blood was sampled three times a day during the inter-ovulatory interval for measuring reproductive and metabolic hormones. On days 9, 11 and 14 of the oestrous cycle, half of the ewes from each group (n = 5) were bled intensively to determine the concentrations of glucose, insulin, IGF-I and leptin. Plasma FSH, progesterone, oestradiol and androstenedione concentrations were similar among groups. Dietary supplementation increased plasma insulin concentrations from the first to the sixth day of supplementation and increased glucose concentrations on the third day, compared with control ewes. Plasma leptin concentrations were higher in STS ewes from the second to the fifth day of supplementation. The pattern of IGF-I concentrations was similar among groups. In STS ewes, the nutritional treatment prolonged the lifespan of the last non-ovulatory follicle, so fewer follicular waves developed during the cycle. In STS ewes, increased concentrations of glucose, insulin and leptin one day before ovulatory wave emergence were associated with increased numbers of follicles growing from 2 to 3 mm and with stimulation of the dominant follicle to grow for a longer period. We suggest that the mechanism by which short-term nutritional supplementation affects follicle development does not involve an increase in FSH concentrations, but may involve responses to increased concentrations of glucose, insulin and leptin, acting directly at the ovarian level. This effect is acute, since concentrations of all three substances decrease after reaching peak values on the third day of supplementation. The status of follicle development at the time of maximum concentrations of glucose and metabolic hormones may be one of the factors that determines whether ovulation rate increases or not.

Effects of feed intake on composition of sheep rumen contents and their microbial population size.

The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed.