A high protein meal affects plasma insulin concentrations and amino acid metabolism in horses with equine metabolic syndrome.

Equine metabolic syndrome (EMS) is characterized by an abnormal insulin response to a glycemic challenge but despite the known insulinotropic effects of certain amino acids, there is a paucity of data evaluating the impact of dietary protein on insulin dynamics in these horses. The objective was therefore to assess insulin and amino acid responses following intake of a high protein meal in healthy horses and those with EMS. Six mature horses diagnosed with EMS and six age-matched control horses without EMS were used. Horses were fed 2g/kg body mass (BM) of a high protein pellet (31% crude protein) at time 0 and 30min, for a total of 4g/kg BM, following an overnight fast. Blood samples collected during a 4h period were analysed for plasma glucose, insulin, amino acids and urea concentrations. Glucose concentrations were not different between groups (P=0.2). Horses with EMS had a 9-fold greater insulinemic response to the consumption of a high protein meal compared with controls (P=0.046). Post-prandial levels of histidine, citrulline, tyrosine, valine, methionine, isoleucine, leucine and ornithine were higher in horses with EMS (P<0.05). Baseline urea nitrogen concentrations were not significantly different between groups (P=0.1). Knowing that certain amino acids are insulin secretagogues, these results illustrate that consumption of a high protein meal caused a hyperinsulinemic response and affected amino acid dynamics in horses with EMS. These findings suggest that dietary protein content should be taken into consideration in the management of horses with insulin dysregulation.

A novel objective chute score interacts with monensin to affect growth of receiving cattle.

Growth in cattle may be related to animal temperament via alterations in intake or feed conversion. However, temperament is ill-defined, and different temperament measures may relate differently to production traits or interact with dietary factors in their effects. To examine relationships between diet, temperament, growth, and health, 160 crossbred steers (262 ± 22 kg) were used in a 56-d RCBD experiment with a 2 × 2 × 2 factorial treatment structure with 5 pens/treatment. Steers were pen fed a corn silage-based diet with or without monensin (41.9 g/t DM), ad libitum. Temperament treatments (assigned on d -7) were exit velocity (EV; slow vs. fast) and objective chute score (OCS; low vs. high), a novel temperament measure, representing the CV of weights collected at 5 measures/s for 10 s while an animal's head was restrained in a chute. Both were measured on d -7, 0, 14, 28, 55, and 56. Subjective chute scores (SCS; visual estimates of animal activity obtained simultaneously with OCS measures) were measured on d -7 and d 56. Jugular blood samples from d 28 were analyzed for antibody response to leptospirosis vaccine and NEFA concentrations. No monensin × OCS × EV interactions were detected ( ≥ 0.11). There was a positive correlation between SCS and OCS ( < 0.01; = 0.57). Changes in OCS and EV across the duration of the study differed among treatments (treatment × day, < 0.10) and indicated that initial measures may be better proxies of growth than average measures. There were no interactions between EV and OCS ( ≥ 0.15) for any response variable and no interactions among treatments ( ≥ 0.31), nor main effects of temperament factors ( ≥ 0.12) for DMI (%BW). Monensin decreased DMI ( < 0.01) similarly across all levels of EV and OCS. Gains and G:F responses to monensin depended on OCS ( < 0.10) but not EV ( ≥ 0.80). Gain was reduced ( < 0.10) by monensin with low, but not high, OCS, and G:F was increased ( < 0.10) by monensin on high, but not low, OCS. Gain during the second 4 wk was lesser ( = 0.04) in fast, compared with slow, EV animals. Results provide novel indications that certain temperament measures can interact with dietary manipulation to influence animal performance.

Relationships of a novel objective chute score and exit velocity with growth performance of receiving cattle.

Animals with excitable temperaments often have decreased gains that have been associated with decreased intake and efficiency. Different temperament measures probably measure different specific underlying traits. Commonly used temperament measures include both objective and subjective measures. Subjective measures present potential difficulties for making across-study comparisons and thus for generalizing quantitative relationships. One objective of this experiment was to evaluate 2 related, but different, measures associated with temperament, where 1 measure is a new, objective measurement based on the common subjective chute score measures. Also, there is reason to believe that RDP requirements of animals may vary with temperament. To examine the relationships between temperament measures and nutrient use, 192 crossbred steers were used in a 58-d randomized complete block design experiment. Temperament treatments (assigned prior to d 1) were chute exit velocity (EV; slow vs. fast) and objective chute score (WSD; low vs. high), a novel temperament measure that was the SD of weights collected at 5 Hz for 10 s while an animal was restrained in a chute with its head caught. Both were measured on d -8, 1, 2, 16, 30, 56, and 58, where d 1 was the day that animals were allotted to treatment groups and began receiving experimental diets. Steers were fed a diet with 1 of 3 RDP levels (75%, 105%, and 120% of RDP requirements). There were no main effects or interactions with RDP ( ≥ 0.12); thus, it was removed from the statistical model for subsequent analyses. There were no interactions between EV and WSD ( ≥ 0.11). Slow EV animals had greater ADG ( = 0.02) and DMI ( ≤ 0.09) than fast EV animals, but there was no effect of EV on G:F ( > 0.14). For d 0 to 58, high WSD animals had greater DMI ( ≤ 0.09) than low WSD animals but no difference in ADG ( = 0.23), whereas low WSD animals tended to have increased G:F ( = 0.11). Results of this study give additional confirmation that EV is associated with DMI and growth and provide evidence that a novel measure of behavior, WSD, is also related to growth, independently of EV. Because WSD and EV appear to measure different underlying behavioral traits, use of both measures may improve our ability to discriminate among temperament categories for growing cattle.

Direct-fed microbials containing lactate-producing bacteria influence ruminal fermentation but not lactate utilization in steers fed a high-concentrate diet.

Direct-fed microbials (DFM) have been shown to improve gain and growth efficiency and also modulate ruminal fermentation. In Exp. 1,72 beef steers were used to compare a lactate-producing bacterial (LAB) DFM consisting primarily of Lactobacillus acidophilus and Enterococcus faecium,and a lactate-producing and lactate-utilizing (LAB/LU) DFM consisting primarily of L. acidophilus and Propionibacterium both fed at 10(9) cfu/d. Steers were fed a corn-based finishing diet for 153 d and then slaughtered for collection of carcass characteristics. In Exp. 2, 12 ruminally cannulated steers were fed acorn-based finishing diet and treated with 10(9) cfu/d of LAB DFM. Rumen fluid was sampled on d 14 and 28 over a 12-h period. Steers were ruminally dosed with a 2-L solution of neutralized DL-lactate (0.56 M)and Cr-EDTA (13.22 M) 3 h postfeeding on d 15 and 29. Ruminal samples were collected at 10- and 20-minintervals for the first and second hour postdosing. No differences (P ≥ 0.14) between control (CON) and LAB for DMI, ADG, growth efficiency, or carcass characteristics were observed. Dry matter intake was greater (P = 0.04) for LAB/LU than LAB from d 0 to 28 but did not differ (P ≥ 0.29) thereafter. Average daily gain was greater (P = 0.04) and efficiency tended(P = 0.06) to be greater for LAB than LAB/LU over the entire 153 d. In Exp. 2, total VFA concentration and molar proportions of butyrate were unaffected(P ≥ 0.24). Molar proportions of acetate exhibited a DFM by hour interaction (P = 0.04); however, on average, molar proportion of acetate was 4.4% greater for DFM. Conversely, DFM did not affect the molar proportion of propionate (P = 0.39). On average,molar proportions of propionate tended to increase(P = 0.07), and acetate tended to decrease (P = 0.07)across days. Mean daily ruminal pH was similar for CON on d 14 and 28, whereas mean pH increased from d 14 to 28 for DFM (DFM × day; P = 0.08).Minimum pH remained unchanged for CON over time but increased from d 14 to 2 for DFM (DFM × day;P = 0.10). Maximum pH decreased from d 14 to 28 in CON but increased over time with DFM (DFM × day;P = 0.05). DL- and L-lactate utilization were unaffected by DFM (P ≥ 0.33) or day (P ≥ 0.50). Although the LAB DFM did not impact growth performance, itd id modulate ruminal fermentation, as evidenced by shifts in ruminal VFA profile and pH; however, DFM did not appear to influence ruminal lactate utilization.

Comparison of in vitro digestibility estimates using the DaisyII incubator with in vivo digestibility estimates in horses.

The objective of this study was to determine if in vitro methodologies developed for the Ankom Daisy(II) incubator could produce accurate estimates of in vivo equine DM digestibility (DMD) and NDF digestibility (NDFD) when equine feces were used as the inoculum source. Four mature geldings were utilized in a 4 × 4 Latin square design experiment with a 2 × 2 factorial arrangement of dietary treatments (timothy hay, alfalfa hay, timothy hay plus oats, and alfalfa hay plus oats), in which the geldings were individually housed and fed. During each 5-d total fecal collection period, feces were collected and composited daily and used to calculate in vivo digestibility. Digestion of the 4 treatment diets was evaluated in vitro using the Daisy(II) incubator. Each incubation vessel of the Daisy(II) was assigned to 1 of the horses and contained 18 filter bags (6 containing the assigned treatment hay, 6 containing hay-oat mix, and 6 containing oats). Three incubation periods were evaluated: 30, 48, and 72 h. Although the 30- and 48-h in vitro estimates were consistently less than the in vivo estimates, they ranked diets in the same order as the in vivo method. For the alfalfa oat diet, timothy diet, and the timothy oat diet, the mean 72-h in vitro DMD and in vivo DMD were not different (P = 0.1444). However, for the alfalfa diet, the DMD estimate from 72-h in vitro incubation was less than the in vivo estimate (P < 0.010). For NDFD, the timothy diet was the only diet, in which the mean 72-h in vitro NDFD estimate was not different than the in vivo estimate. However, the in vitro method correctly ranked the alfalfa-based diets as having greater NDFD estimates than the timothy-based diets. Of the 3 incubation periods, the 72-h period provided digestibility estimates most similar to the in vivo data. Using the methodologies described in this research, the Daisy(II) incubator and equine feces can be used to estimate in vivo DMD of horse feeds.

The small intestinal epithelia of beef steers differentially express sugar transporter messenger ribonucleic acid in response to abomasal versus ruminal infusion of starch hydrolysate.

In mammals, the absorption of monosaccharides from small intestinal lumen involves at least 3 sugar transporters (SugT): sodium-dependent glucose transporter 1 (SGLT1; gene SLC5A1) transports glucose and galactose, whereas glucose transporter (GLUT) 5 (GLUT5; gene SLC2A5) transports fructose, across the apical membrane of enterocytes. In contrast, GLUT2 (gene SLC2A2) transports all of these sugars across basolateral and apical membranes. To compare the distribution patterns and sensitivity with nutritional regulation of these 3 SugT mRNA in beef cattle small intestinal tissue, 18 ruminally and abomasally catheterized Angus steers (BW approximately 260 kg) were assigned to water (control), ruminal cornstarch (partially hydrolyzed by alpha-amylase; SH), or abomasal SH infusion treatments (n = 6) and fed an alfalfa-cube-based diet at 1.3 x NE(m) requirement. The SH infusions amounted to 20% of ME intake. After 14- or 16-d of infusion, steers were killed; duodenal, jejunal, and ileal epithelia harvested; and total RNA extracted. The relative amount of SugT mRNA in epithelia was determined using real-time reverse transcription-PCR quantification methods. Basal expression of GLUT2 and SGLT1 mRNA was greater (P < 0.09) by jejunal than by duodenal or ileal epithelia, whereas basal content of GLUT5 mRNA was greater (P < or = 0.02) by jejunal and duodenal than by ileal epithelia. The content of GLUT5 mRNA in small intestinal epithelia was not affected (P > or = 0.16) by either SH infusion treatment. In contrast, GLUT2 and SGLT1 mRNA content in the ileal epithelium was increased (P < or = 0.05) by 6.5- and 1.3-fold, respectively, after abomasal SH infusion. Duodenal SGLT1 mRNA content also was increased (P = 0.07) by 64% after ruminal SH infusion. These results demonstrate that the ileum of beef cattle small intestine adapts to an increased luminal supply of glucose by increasing SGLT1 and GLUT2 mRNA content, whereas increased ruminal SH supply results in duodenal upregulation of SGLT1 mRNA content. These adaptive responses of GLUT2 and SGLT1 mRNA to abomasal or ruminal SH infusion suggest that beef cattle can adapt to increase their carbohydrate assimilation through small intestinal epithelia, assuming that altered SugT mRNA contents reflect the altered transport functional capacities.

Ruminal and abomasal starch hydrolysate infusions selectively decrease the expression of cationic amino acid transporter mRNA by small intestinal epithelia of forage-fed beef steers.

Although cationic amino acids (CAA) are considered essential to maximize optimal growth of cattle, transporters responsible for CAA absorption by bovine small intestinal epithelia have not been described. This study was conducted to test 2 hypotheses: 1) the duodenal, jejunal, and ileal epithelia of beef cattle differentially express 7 mRNA associated with 4 mammalian amino acid (AA) transport activities: y(+) (CAT1), B(0,+) (ATB(0,+)), b(0,+) (b(0,+)AT and rBAT), and y(+)L (y(+)LAT1, y(+)LAT2, and 4F2hc), and 2) the expression of these mRNA is responsive to small intestinal luminal supply of AA substrates (derived from ruminal microbes) or glucose-derived energy (from starch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (body weight = 260 +/- 17 kg) fed an alfalfa cube-based diet at 1.33 x net energy for maintenance requirement were assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control); ruminal SH and abomasal water infusion; and ruminal water and abomasal SH infusion. The dosage of SH infusion amounted to 20% of metabolizable energy intake. After 14 or 16 d of infusion, steers were slaughtered, duodenal, jejunal, and ileal epithelia were harvested, and total RNA was extracted. The relative amounts of mRNA expressed by epithelia were quantified using real-time reverse transcription-PCR. All 7 mRNA species were expressed by the epithelium from each region, but their abundance differed among the regions. Specifically, duodenal expression of CAT1 and ATB(0,+) mRNA was greater than jejunal or ileal expression; ileal expression of b(0,+)AT, rBAT, and y(+)LAT1 mRNA was greater than jejunal or duodenal expression, whereas the expression of y(+)LAT2 and 4F2hc mRNA did not differ among the 3 epithelia. With regard to SH infusion effect, ruminal infusion down-regulated or tended to down-regulate the jejunal expression of CAT1, rBAT, y(+)LAT2, and 4F2hc mRNA. Abomasal infusion down-regulated the jejunal expression of y(+)LAT2 mRNA and tended to down-regulate the jejunal expression of 4F2hc mRNA. This study characterized the pattern of CAA transporter mRNA expressed by growing beef cattle fed an alfalfa-based diet. Moreover, this study demonstrated that increasing the luminal supply of microbe-derived AA (by ruminal supplementation of SH) results in a reduced capacity of apical and basolateral membrane to transport of CAA, whereas increasing luminal glucose supply (by abomasal supplementation of SH) reduces only the basolateral transport capacity, assuming that CAA transporter mRNA content represents functional capacity.

Influence of slow-release urea on nitrogen balance and portal-drained visceral nutrient flux in beef steers.

Two experiments were conducted to evaluate the effects of slow-release urea (SRU) versus feed-grade urea on portal-drained visceral (PDV) nutrient flux, nutrient digestibility, and total N balance in beef steers. Multi-catheterized steers were used to determine effects of intraruminal dosing (Exp. 1; n = 4; 319 +/- 5 kg of BW) or feeding (Exp. 2; n = 10; 4 Holstein steers 236 +/- 43 kg of BW and 6 Angus steers 367 +/- 46 kg of BW) SRU or urea on PDV nutrient flux and blood variables for 10 h after dosing. Intraruminal dosing of SRU (Exp. 1) prevented the rapid increase in ruminal ammonia concentrations that occurred with urea dosing (treatment x time P = 0.001). Although apparent total tract digestibilities of DM, OM, NDF, and ADF were not affected by treatment (P > 0.53, Exp. 2), SRU increased fecal N excretion (49.6 vs. 45.6 g/d; P = 0.04) and reduced apparent total tract N digestibility (61.7 vs. 66.0%; P = 0.003). Transfer of urea from the blood to the gastrointestinal tract occurred for both treatments in Exp. 1 and 2 at all time points with the exception for 0.5 h after dosing of urea in Exp. 1, when urea was actually transferred from the gastrointestinal tract to the blood. In both Exp. 1 and 2, both urea and SRU treatments increased arterial urea concentrations from 0.5 to 6 h after feeding, but arterial urea concentrations were consistently less with SRU (treatment x time P < 0.001, Exp. 1; P = 0.007, Exp. 2). Net portal ammonia release remained relatively consistent across the entire sampling period with SRU treatment, whereas urea treatment increased portal ammonia release in Exp. 1 and tended to have a similar effect in Exp. 2 (treatment x time P = 0.003 and P = 0.11, respectively). Urea treatment also increased hepatic ammonia uptake within 0.5 h (treatment x time P = 0.02, Exp. 1); however, increased total splanchnic release of ammonia for the 2 h after urea treatment dosing suggests that PDV ammonia flux may have exceeded hepatic capacity for removal. Slow-release urea reduces the rapidity of ammonia-N release and may reduce shifts in N metabolism associated with disposal of ammonia. However, SRU increased fecal N excretion and increased urea transfer to the gastrointestinal tract, possibly by reduced SRU hydrolysis or effects on digestion patterns. Despite this, the ability of SRU to protect against the negative effects of urea feeding may be efficacious in some feeding applications.

Effects of slow-release urea on ruminal digesta characteristics and growth performance in beef steers.

Two experiments were conducted to evaluate the effects of slow-release urea (SRU) versus feed-grade urea on ruminal metabolite characteristics in steers and DMI, gain, and G:F in growing beef steers. Experiment 1 used 12 ruminally cannulated steers (529 +/- 16 kg of BW) to monitor the behavior of SRU in the ruminal environment. Compared with feed-grade urea, SRU decreased ruminal ammonia concentration (P = 0.02) and tended to increase ruminal urease activity (P = 0.06) without affecting ruminal VFA molar proportions or total concentrations (P > 0.20). After 35 d of feeding, the in situ degradation rate of SRU was not different between animals fed urea or SRU (P = 0.48). Experiment 2 used 180 Angus-cross steers (330 +/- 2.3 kg) fed corn silage-based diets supplemented with urea or SRU for 56 d to evaluate the effects on feed intake, gain, and G:F. The design was a randomized complete block with a 2 x 4 + 1 factorial arrangement of treatments. Treatments included no supplemental urea (control) or urea or SRU at 0.4, 0.8, 1.2, or 1.6% of diet DM. Over the entire 56 d experiment, there were interactions of urea source x concentration for gain (P = 0.04) and G:F (P = 0.01) because SRU reduced ADG and G:F at the 0.4 and 1.6% supplementation concentrations but was equivalent to urea at the 0.8 and 1.2% supplementation concentrations; these effects were due to urea source x concentration interactions for gain (P = 0.06) and G:F (P = 0.05) during d 29 to 56 of the experiment. The SRU reduced DMI during d 29 to 56 (P = 0.01) but not during d 0 to 28, so that over the entire experiment there was no difference in DMI for urea source (P = 0.19). These collective results demonstrate that SRU releases N slowly in the rumen with no apparent adaptation within 35 d. Supplementation of SRU may limit N availability at low (0.4%) concentrations but is equivalent to urea at 0.8 and 1.2% concentrations.

Nutrient net absorption across the portal-drained viscera of forage-fed beef steers: Quantitative assessment and application to a nutritional prediction model.

This study aimed to establish the relationship between ME intake and energy and nutrient absorption across the portal-drained viscera (PDV) of forage-fed beef steers. Eight Angus (328 +/- 40 kg of BW) steers were surgically fitted with portal, mesenteric arterial, and mesenteric venous catheters, and were fed alfalfa cubes in a replicated 4 x 4 Latin square design with 4 levels of energy intake between 1 and 2 times maintenance energy requirements. On d 28 of each experimental period, p-aminohippuric acid was infused to measure blood and plasma flow across the PDV, and blood samples (1 every hour, for 6 h) were collected simultaneously from arterial and venous catheters for net absorption measurements. Oxygen utilization, and therefore energy utilization, increased (P < 0.05) linearly in relation to ME intake. Glucose net uptake was unaffected, but lactate net release increased linearly in response to ME intake (P < 0.05). Net absorption of all AA except tryptophan, glutamate, and glutamine increased linearly with ME intake (P < 0.05). The constant net absorption of glutamate and glutamine indicated increased net utilization of these AA when dietary supply was increased. These data provide quantitative measures of the PDV effects on energy and AA availability for productive tissues, and suggest that the greater net utilization of some AA when ME intake is increased could relate to their catabolism for energy production. Prediction estimates of small intestinal AA absorption, based on the Cornell Net Carbohydrate and Protein System (CNCPS), exceeded observed net AA PDV absorption. Mean bias represented the greatest proportion (87 to 96%) of the deviation between individual AA absorption and observed net AA PDV absorption, suggesting that the CNCPS model may be used to predict AA net absorption when factors describing AA utilization by the PDV are applied to model predictions.

Basal expression of nucleoside transporter mRNA differs among small intestinal epithelia of beef steers and is differentially altered by ruminal or abomasal infusion of starch hydrolysate.

In ruminants, microbial-derived nucleic acids are a major source of N and are absorbed as nucleosides by small intestinal epithelia. Although the biochemical activities of 2 nucleoside transport systems have been described for cattle, little is known regarding the regulation of their gene expression. This study was conducted to test 2 hypotheses: (1) the small intestinal epithelia of beef cattle differentially express mRNA for 3 concentrative (CNT1, 2, 3) and 2 equilibrative (ENT1, 2) nucleoside transporters (NT), and (2) expression of these NT is responsive to small intestine luminal supply of rumen-derived microbes (hence, nucleosides), energy (cornstarch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (260 +/- 17 kg of BW) were fed an alfalfa cube-based diet at 1.33x NE(m) requirement. Six steers in each of 3 periods were blocked by BW (heavy vs. light). Within each block, 3 steers were randomly assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control), ruminal SH infusion/abomasal water infusion, or ruminal water infusion/abomasal SH infusion. The dosage of SH infusion amounted to 20% of ME intake. After a 14-or 16-d infusion period, steers were slaughtered, and duodenal, jejunal, and ileal epithelia were harvested for total RNA extraction and the relative amounts of mRNA expressed were determined using real-time RT-PCR quantification methodologies. All 5 NT mRNA were found expressed by each epithelium, but their abundance differed among epithelia. Specifically, jejunal expression of all 5 NT mRNA was higher than that by the ileum, whereas jejunal expression of CNT1, CNT3, and ENT1 mRNA was higher, or tended to be higher, than duodenal expression. Duodenal expression of CNT2, CNT3, and ENT2 mRNA was higher than ileal expression. With regard to SH infusion treatments, ruminal infusion increased duodenal expression of CNT3 (67%), ENT1 (51%), and ENT2 (39%) mRNA and ileal expression of CNT3 (210%) and ENT2 (65%) mRNA. Abomasal infusion increased (54%) ileal expression of ENT2 mRNA and tended to increase (50%) jejunal ENT2 mRNA expression. This study has uniquely characterized the pattern of NT mRNA expression by growing beef cattle and found that the mRNA abundance for CNT3, ENT1, and ENT2 in small intestinal epithelia can be increased by increasing the luminal supply of nucleotides (CNT3, ENT1, ENT2) or glucose (ENT2).

Identification and expression pattern of cationic amino acid transporter-1 mRNA in small intestinal epithelia of Angus steers at four production stages.

Although dietary supplementation of cationic AA (CAA), especially l-Lys, is known to be essential for optimal growth of beef cattle, the proteins responsible for absorption of CAA by bovine intestinal epithelia have not been described. Cationic AA transporter-1 (CAT-1) is a major intestinal CAA transporter, demonstrating a high-affinity (muM) transport activity for l-Lys in other mammals, and is widely expressed by small intestinal epithelia of nonruminants, but neither sequence nor expression pattern data exist for CAT-1 in cattle. Therefore, the goal of this research was to compare the relative expression (putative) of CAT-1 mRNA by duodenal, jejunal, or ileal small intestinal epithelia across and within commercially relevant beef cattle production and development stages. Twenty-four Angus steers were assigned randomly (n = 6) to 1 of 4 treatments (suckling, weanling, growing, and finishing) after all steers were born. Duodenal, jejunal, and ileal epithelia were scraped, and total RNA was extracted after the steers were killed at 32, 184, 248, or 423 d of age. Average daily gains of the steers did not differ (1.09 +/- 0.05 kg/d) among stages, whereas the small intestinal length relative to BW decreased (P < 0.01) with steer development. Using standard reverse transcription-PCR cloning techniques, we generated a partial-length bovine CAT-1 complementary DNA (695 bp; GenBank accession no. DQ399522) from jejunal mRNA samples, which possessed 89 and 87% identities to pig and human CAT-1 orthologs, respectively. On the basis of this bovine-specific genetic data, a real-time PCR-based assay of reverse-transcribed mRNA was developed and used to measure relative changes in bovine CAT-1 mRNA abundance in intestinal epithelia as steers developed. The CAT-1 mRNA was expressed by the duodenum, jejunum, and ileum of all 4 production stages. In contrast to expression by duodenal or ileal epithelium, jejunal expression of CAT-1 mRNA by growing steers was greater (P = 0.005) than that by suckling, weanling, or finishing steers. In terms of the expression of CAT-1 mRNA within production stage, jejunal expression was greater (P = 0.002) than that by duodenum or ileum for growing steers. In contrast, no intestinal site difference was found for suckling, weanling, or finishing steers. These data indicate that previously reported Na(+)-independent uptake of Lys by jejunal and ileal epithelia likely occurred by CAT-1, and that the potential capacity for CAT-1-mediated uptake of CAA for beef steers may be greatest for the "growing" phenotype.

Evaluation of perennial ryegrass straw as a forage source for ruminants.

Two experiments were conducted to evaluate perennial ryegrass straw as a forage source for ruminants. Experiment 1 evaluated digestion and physiological variables in steers offered perennial ryegrass straw containing increasing levels of ergot alkaloid, lolitrem B. Sixteen ruminally cannulated Angus x Hereford steers (231+/-2 kg BW) were blocked by weight and assigned randomly to one of four treatments. Steers were provided perennial ryegrass straw at 120% of the previous 5-d average intake. Before straw feeding, soybean meal was provided (0.1% BW; CP basis) to meet the estimated requirement for degradable intake protein. Low (L) and high (H) lolitrem B straws (<100 and 1,550 ppb, respectively; DM basis) were used to formulate treatment diets: 100% L; 67% L:33% H; 33% L:67% H; 100% H (DM basis). Intake and digestibility of DM and OM, and ruminal pH, total VFA, and NH3-N were not affected by increasing lolitrem B concentration. Ruminal indigestible ADF (IADF) fill increased linearly (P = 0.01) and IADF passage rate decreased linearly (P = 0.04) as lolitrem B increased. Experiment 2 evaluated performance and production by 72 Angus x Hereford cows (539+/-5 kg BW) consuming perennial ryegrass straw containing increasing lolitrem B during the last third of gestation. Cows were blocked by body condition score and randomly assigned to one of three treatments. Cows were provided perennial ryegrass straw ad libitum and supplemented with soybean meal (0.1% BW; CP basis) to meet the estimated requirement for degradable intake protein. Mixtures of a L and H lolitrem B straw (467 and 2,017 ppb, respectively) were used to formulate treatment diets: 100% L, 50% L:50% H, 100% H (DM basis). Thirteen of 24 cows on the 100% H treatment exhibited signs of ryegrass staggers and were removed from the study; nevertheless, lolitrem B concentration did not influence pre- or postcalving weight or body condition score change. These data suggest that feeding perennial ryegrass straw containing up to 1,550 ppb lolitrem B (DM basis) did not adversely affect nutrient digestion or physiological response variables in steers. However, providing straw with a lolitrem B concentration of approximately 2,000 ppb (DM basis) resulted in 54% of cows exhibiting signs of ryegrass staggers. These data suggest that blending straws with a high (>2,000 ppb) and low (<500 ppb) concentration of lolitrem B can be a successful management practice.

Interactions between supplement energy source and tall fescue hay maturity on forage utilization by beef steers.

This experiment was conducted to determine the effects of tall fescue hay maturity on intake, digestion, and ruminal fermentation responses to different supplemental energy sources fed to beef steers. Twelve ruminally cannulated, crossbred steers (initial BW = 228 +/- 21 kg) were used in a split-plot experiment with a 3 x 4 factorial treatment arrangement. Steers were assigned randomly to three supplement treatments: 1) no supplement, 2) pelleted soybean hulls, or 3) coarse cracked corn. The second treatment factor was fescue hay maturity: 1) vegetative (VEG), 2) boot-stage (BOOT), 3) heading-stage (HEAD), and 4) mature (MAT). Supplements were fed once daily at 0.67% of BW (OM basis) and tall fescue hay was offered once daily at 150% of average intake. Supplement type x forage maturity interactions were not detected (P > or = 0.25) for forage, total, or digestible OM intake, which generally decreased (P < 0.01) with advancing forage maturity. Supplementation decreased (P < 0.01) forage and increased (P < 0.01) total OM intake. Supplement type had no effect (P = 0.56) on substitution ratio (unit change in forage intake per unit of supplement intake). Digestible OM intake was increased (P < 0.01) by supplementation and was greater (P = 0.05) with soybean hulls than with corn. Supplement type x forage maturity interactions (P < or = 0.10) were observed for OM and NDF digestibilities and N retention. Increases in digestibility with soybean hulls relative to corn were greater and supplementation elicited greater increases in N retention with more mature forages. Compared with soybean hulls, corn supplementation resulted in greater (P < 0.01) negative associative effects on OM digestibility. Supplementation did not affect (P > or = 0.10) ruminal pH, total VFA concentrations, or acetate:propionate ratio. Corn supplementation decreased (P < or = 0.07) ruminal NH3-N concentrations compared with control and soybean hulls; however, decreases in ruminal NH3-N concentrations were not consistent with the presence of negative associative effects. Thus, mechanisms not involving ruminal pH or NH3-N concentration seem responsible for negative associative effects observed with corn supplementation. Within the range of forage quality in this study, increases in digestible OM intake from starch- or fiber-based supplements were independent of forage maturity. When fed at similar levels of OM, soybean hull supplementation provided an average of 6% greater digestible OM intake than corn supplementation.

Effect of supplemental nutrient source on heifer growth and reproductive performance, and on utilization of corn silage-based diets by beef steers.

Two experiments were conducted to determine effects of oilseeds or soybean hulls on growth and reproductive performance of heifers and utilization of corn silage diets by growing beef cattle. In Exp. 1, 96 beef heifers (249 kg of BW) were used in a randomized complete block design. Treatments were as follows: 1) corn and soybean meal (CON) at 56% of the DMI; 2) whole linted cottonseed at 15% of the DMI (COT); 3) whole raw soybeans at 15% of the DMI (SB); or 4) pelleted soyhulls at 30% of the DMI (SH). Diets were formulated to be isonitrogenous (13.8% CP) and fed to achieve target weights equal to 65% of expected mature BW at the time of AI. Estrus was synchronized and heifers were inseminated by AI in response to detected estrus. Because the energy value for SH was underestimated, cumulative ADG for SH (1.03 kg/d) was greater (P < or = 0.03) than for CON (0.89 kg/d), COT (0.87 kg/d), or SB (0.86 kg/d). Treatment did not affect (P > 0.10) the proportion of pubertal heifers at the beginning of the breeding season: CON (60%), COT (53%), SB (69%), SH (71%), or first-service conception rates: CON (37%); COT (38%); SB (57%); SH (42%). In Exp. 2, crossbred steers (387 kg) were used in a 6 x 6 Latin square design to evaluate the effects of supplemental nutrient source on utilization of corn silage diets. Treatments included diets used in Exp. 1, plus a negative control (soybean meal at 10% of the DMI; SIL) and whole raw soybeans at 25% of the DMI (SB25). Diets were formulated to be isonitrogenous (13.8% CP) except SB25 (17% CP), and were fed twice daily at 1.8 x NEm. Oilseed inclusion decreased (P < 0.10) acetate:propionate ratios and (P < 0.10) apparent ruminal OM and ruminal and total tract NDF digestibilities. The CON and SH diets had the greatest (P < 0.10) total-tract OM digestibilities. Microbial efficiencies were greatest (P < 0.10), and long chain fatty acid flow to the duodenum increased (P < 0.10) with oilseeds. Biohydrogenation averaged 90.4% and increased slightly (P < 0.10) when oilseeds were added to the diet. Adding oilseeds or soybean hulls to corn silage-based diets did not affect reproductive performance of heifers. Although oilseed additions increased total fatty acid flow to the duodenum, a high degree of biohydrogenation occurred, greatly increasing C18:0, with only marginal increases in unsaturated fatty acid flow. Depending on diet and feeding conditions, inclusion of whole oilseeds may not be an effective means of increasing linoleic acid supply for ruminant animals.

Characterization of season and sampling method effects on measurement of forage quality in fescue-based pastures.

Information on seasonal changes and effects of sampling methods on the measurement of forage quality is limited for fescue-based pastures. Eight continuously grazed, 0.76-ha, fescue-based pastures were used to compare forage type, method of collection, and seasonal effects on forage quality in a repeated-measures, split-plot design. Four pastures were interseeded with red clover in March 2000. Masticate (M; from four ruminally cannulated steers) and hand--clipped (C) samples were collected every 28 d from April to October 2000. Interseeding red clover did not affect (P > 0.10) OM, CP, NDF, and ADF concentrations or CP degradability. Sampling method and season interacted (P < 0.03) for OM, CP, NDF, and ADF concentrations. Concentrations of OM averaged 5 percentage units more (P < 0.01) in C than in M in all months and were more variable with M than with C. Samples clipped between April and September averaged 5.5 percentage units greater NDF (P < 0.01), 3.0 percentage units greater ADF (P < 0.01), and 4.5 percentage units less CP (P < 0.01) than masticate samples obtained during the same time period. Fiber and CP concentrations did not differ (P > 0.10) between C and M samples obtained in October. Differences in CP degradability estimates (using Streptomyces griseus protease) between the two sample types were greater in late-season samples than in samples obtained from April to June. When S. griseus protein degradability estimates were compared with in situ estimates for masticate samples, no differences (P > 0.10) were detected early in the season (April to June). However, the S. griseus procedure overestimated in situ values (P < 0.01) by an average of 3 percentage units in samples obtained between July and October. Differences in composition of C and M samples were substantial until late season, when opportunities for selective grazing were minimal. Small differences between S. griseus and in situ estimates of CP degradability indicate that the S. griseus procedure can yield useful CP degradability estimates for fescue-based pasture samples. Although it might be possible to apply correction values to clipped samples to estimate CP and fiber concentrations of diets selected by grazing cattle, inconsistent relationships preclude this approach for estimates of CP degradability.

Effect of increasing proportion of supplemental N from urea in prepartum supplements on range beef cow performance and on forage intake and digestibility by steers fed low-quality forage.

Four experiments were conducted to evaluate the influence of changing the proportion of supplemental degradable intake protein (DIP) from urea on forage intake, digestion, and performance by beef cattle consuming either low-quality, tallgrass prairie forage (Exp. 1, 2, and 4) or forage sorghum hay (Exp. 3). Experiments 1, 2, and 3 were intended to have four levels of supplemental DIP from urea: 0, 20, 40, and 60%. However, refusal to consume the 60% supplement by cows grazing tallgrass prairie resulted in elimination of this treatment from Exp. 1 and 2. Levels of supplemental DIP from urea in Exp. 4 were 0, 15, 30, and 45%. Supplements contained approximately 30% CP, provided sufficient DIP to maximize digestible OM intake (DOMI) of low-quality forage diets, and were fed to cows during the prepartum period. In Exp. 1, 12 Angus x Hereford steers (average initial BW = 379) were assigned to the 0, 20, and 40% treatments. Forage OM intake, DOMI, OM, and NDF digestion were not affected by urea level. In Exp. 2, 90 pregnant, Angus x Hereford cows (average initial BW = 504 kg and body condition [BC] = 5.0) were assigned to the 0, 20, and 40% treatments. Treatment had little effect on cow BW and BC changes and calf birth weight, ADG, or weaning weight. However, pregnancy rate tended to be lowest (P = 0.13) for the greatest level of urea. In Exp. 3, 120 pregnant, crossbred beef cows (average initial BW = 498 kg and BC = 4.6) were assigned to the 0, 20, 40, and 60% treatments. Prepartum BC change tended (P = 0.08) to be quadratic (least increase for 60% treatment), although BW change was not statistically significant. Treatment effect on calf birth weight was inconsistent (cubic; P = 0.03), but calf ADG and weaning weight were not affected by treatment. Pregnancy rate was not affected by prepartum treatment. In Exp. 4, 132 pregnant, Angus x Hereford cows (average initial BW = 533 and BC = 5.3) were assigned to the 0, 15, 30, and 45% treatments. Prepartum BC loss was greatest (quadratic; P = 0.04) for the high-urea (45%) treatment, although BW loss during this period declined linearly (P < 0.01). Prepartum treatment did not affect pregnancy rate, calf birth weight, or ADG. In conclusion, when sufficient DIP was offered to prepartum cows to maximize low-quality forage DOMI, urea could replace between 20 and 40% of the DIP in a high-protein (30%) supplement without significantly altering supplement palatability or cow and calf performance.

Effects of supplemental degradable intake protein on utilization of medium- to low-quality forages.

Three independent experiments were conducted each using 16 ruminally fistulated beef steers fed bermudagrass (8.2% CP, 71% NDF; Exp. 1), bromegrass (5.9% CP, 65% NDF; Exp. 2), or forage sorghum (4.3% CP, 60% NDF; Exp. 3) hays to evaluate the effects of increasing level of supplemental degradable intake protein (DIP) on forage utilization. In each experiment, steers were blocked by weight and assigned to one of four treatments, and hay was offered to each steer at 130% of average voluntary intake for the preceding 5-d period. Supplemental DIP (sodium caseinate) was placed in the rumen at 0700, immediately before feeding forage. Levels of DIP supplementation were .041, .082, and .124% BW; the control received no supplemental DIP. Following a 10-d adaptation, intake and total fecal output were measured for 7 d. In Exp. 1, neither forage OM intake (FOMI) nor fiber (NDF) digestion were influenced (P > or = .20) by increasing level of DIP supplementation. The DIP supplied by the bermudagrass hay was estimated to be 8.2% of the total digestible OM intake (TDOMI) for control steers. In Exp. 2, increasing level of supplemental DIP did not affect (P > or = .26) FOMI but tended to increase total OM intake linearly (TOMI; P = .10). The tendency for a rise in TOMI coupled with a slight numeric increase in digestion resulted in an increase (linear; P = .06) in TDOMI. In the treatment group in which the maximum TDOMI was observed (supplemental DIP treatment of .082% BW), total DIP intake constituted approximately 9.8% of the TDOMI. In Exp. 3, FOMI, TOMI, organic matter digestion (OMD), and TDOMI were improved (P < .01) by increasing amounts of supplemental DIP. Although there was some evidence of a tendency for a decrease in the magnitude of change in TDOMI in response to increasing DIP supplementation, a clear plateau was not achieved with the levels of supplement provided. When the highest level of supplemental DIP was fed, DIP constituted approximately 12.8% of the TDOMI. In conclusion, significant variation was observed among forage in the amount of DIP needed to maximize intake and digestion when expressed in relationship to the digestible OM.

Effects of level and source of carbohydrate and level of degradable intake protein on intake and digestion of low-quality tallgrass-prairie hay by beef steers.

Ruminally fistulated steers (n = 13; 263 kg) were used in an incomplete Latin square with 13 treatments and four periods to evaluate the effects of level and source of supplemental carbohydrate (CHO) and level of degradable intake protein (DIP) on the utilization of low-quality, tallgrass-prairie hay. Steers were given ad libitum access to forage (5.7% CP, 2.6% DIP, and 74.9% NDF). The supplementation treatments were fashioned as a 2x3x2 factorial arrangement plus a negative control (NC; no supplement). The factors included two DIP levels (.031 and .122% BW) and three CHO sources (starch, glucose, and fiber) fed at two levels (.15 and .30% BW) within each level of DIP supplementation. The effect of supplementation on forage OM intake (FOMI) was dependent (P<.01) on level and source of CHO and level of DIP fed. When DIP was low, forage, total, and digestible OM intakes were generally greater for the starch treatment than for the nonstarch treatments. However, when the DIP level was high, intakes were greater for the nonstarch (i.e., fiber and glucose) treatments. Generally, FOMI decreased (P<.01) when more supplemental CHO was provided. Supplementation typically increased fiber digestion, but the response was dependent (P<.01) on level and source of CHO and level of DIP. Generally, supplements with low levels of CHO improved NDF digestion (NDFD). However, supplements with the high level of CHO decreased NDFD, except for fiber at the high level of DIP. Organic matter digestion was increased by supplementation, but the impact of increasing CHO was dependent (P<.01) on source of CHO and level of DIP. Supplementation treatments had significant impact on ruminal pH, NH3 N, and the total concentration of organic acids as well as their relative proportions. In conclusion, supplemental DIP enhanced the use of low-quality forage; however, the impact of supplemental CHO on low-quality forage use was dependent on source and level of CHO offered, as well as the level of DIP provided.

Effects of ruminal administration of supplemental degradable intake protein and starch on utilization of low-quality warm-season grass hay by beef steers.

Hereford x Angus steers were used in a 13-treatment, four-period, incomplete Latin square design to examine the effects of starch and degradable intake protein (DIP) supplements on forage utilization and ruminal function. Steers were given ad libitum access to low-quality hay (4.9% CP) and were not supplemented (NS) or received different amounts of starch (cornstarch grits; 0, .15, and .3% of initial BW) and DIP (Na-caseinate; .03, .06, .09, and .12% of initial BW) administered via ruminal fistulae in a 3 x 4 factorial arrangement of treatments. Supplemented steers consumed more (P < .01) forage OM, total OM, NDF, and digestible OM (DOM) than NS steers. Forage OM, total OM, NDF, and DOM intakes increased linearly (P < .01) as the amount of supplemental DIP increased. The addition of starch to supplements linearly decreased ( P < .01) the intake of forage OM, NDF, and DOM. The digestion of DM, OM, and NDF increased linearly (P < .01) with supplemental DIP and decreased linearly (P < or = .06) with supplemental starch. Particulate and liquid passages generally increased with DIP; however, starch level influenced the nature of the response (P = .03 and .06, respectively). Similarly, ruminal acid detergent-insoluble ash content generally decreased as starch increased, but the effect was dependent on DIP level (P < .01). Supplementation increased (P < .01) ruminal NH3 and total VFA and decreased (P < .01) ruminal pH relative to NS. All treatments supported average pH values in a range (6.3 to 6.7) unlikely to inhibit fibrolytic bacteria. Ruminal NH3 concentration increased quadratically (P = .03) with DIP and decreased linearly (P = .02) with starch. As DIP increased, total VFA concentration increased linearly (P = .02). Providing supplemental DIP to steers fed low-quality forage increased OM intake and digestion, whereas addition of starch to supplements decreased forage intake and digestion.