Search

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 (µM) 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 i 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 Natindependent 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. [ABSTRACT FROM AUTHOR]

Aged beef cows (= 8 yr of age) produce calves with lower birth and weaning weights. In mammals, aging is associated with reduced hepatic expression of glutamine synthetase (GS) and alanine transaminase (ALT), thus impaired hepatic Gln-Glu cycle function. To determine if the relative protein content of GS, ALT, aspartate transaminase (AST), glutamate transporters (EAAC1, GLT-1), and their regulating protein (GTRAP3-18) differed in biopsied liver tissue of (a) aged vs. young (3 to 4 yr old) nonlactating, nongestating Angus cows (Exp. 1 and 2) and (b) aged mixed-breed cows with and without COMPUDOSE (17β-estradiol) ear implants (Exp. 3), Western blot analyses were performed. In Exp. 1, 12 young (3.62 ± 0.01 yr) and 13 aged (10.08 ± 0.42 yr) cows grazed the same mixed forage for 42 d (August-October). In Exp. 2, 12 young (3.36 ± 0.01 yr) and 12 aged (10.38 ± 0.47 yr) cows were individually fed (1.03% of BW) a corn-silage-based diet to maintain BW for 20 d. For both Exp. 1 and 2, the effect of cow age was assessed by ANOVA using the MIXED procedure of SAS. Cow BW did not change (P = 0.17). Hepatic ALT (78% and 61%) and GS (52% and 71%) protein content (Exp. 1 and 2, respectively) was decreased (P = 0.01), whereas GTRAP3-18 (an inhibitor of EAAC1 activity) increased (P = 0.01; 170% and 136%) and AST, GLT-1, and EAAC1 contents did not differ (P = 0.17) in aged vs. young cows. In Exp. 2, free concentrations (nmol/g) of Glu, Ala, Gln, Arg, and Orn in liver homogenates were determined. Aged cows tended to have less (P = 0.10) free Gln (15.0%) than young cows, whereas other AA concentrations did not differ (P = 0.26). In Exp. 3, 14 aged (> 10 yr) cows were randomly allotted (n = 7) to sham or COMPUDOSE (25.7 mg of 17β-estradiol) implant treatment (TRT), and had ad libitum access to alfalfa hay for 28 d. Blood and liver biopsies were collected 14 and 28 d after implant treatment. Treatment, time after implant (DAY), and TRT × DAY effects were assessed by ANOVA using the MIXED procedure of SAS. Cow BW was not affected (P = 0.96). Implant increased (P = 0.02) total plasma estradiol by 220% (5.07 vs. 1.58 pg/mL) and GS protein by 300%, whereas the relative content of other proteins was not altered (P = 0.16). We conclude that hepatic expression of ALT and GS are reduced in aged vs. young cows, and administration of 17β-estradiol to aged cows increases plasma estradiol and hepatic GS, but not that of other proteins that support hepatic Glu metabolism. [ABSTRACT FROM AUTHOR]

As part of a 2-yr study documenting the physiologic impact of grazing endophyte-infected tall fescue on growing cattle, 2 experiments were conducted to characterize and evaluate effects of grazing 2 levels of toxic endophyte-infected tall fescue pastures on vascular contractility and serotonin receptors. Experiment 1 examined vasoconstrictive activities of 5-hydroxytryptamine (5HT), α-methylserotonin (ME5HT; a 5HT2 receptor agonist), D-lysergic acid (LSA), and ergovaline (ERV) on lateral saphenous veins collected from steers immediately removed from a high-endophyte-infected tall fescue pasture (HE) or a low-endophyte-infected mixed-grass (LE) pasture. Using the same pastures, Exp. 2 evaluated effects of grazing 2 levels of toxic endophyte-infected tall fescue on vasoconstrictive activities of (±)-1-(2 ,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI), BW 723C86 (BW7), CGS-12066A (CGS), and 5-carboxamidotryptamine hemiethanolate maleate (5CT), agonists for 5HT2A, 2B, 5HT1B, and 5HT7 receptors, respectively. One-half of the steers in Exp. 2 were slaughtered immediately after removal from pasture, and the other one-half were fed finishing diets for >91 d before slaughter. For Exp. 1, maximal contractile intensities were greater (P < 0.05) for steers grazing LE pastures than HE pastures for 5HT (73.3 vs. 48.9 ± 2.1%), ME5HT (52.7 vs. 24.9 ± 1.5%), and ERV (65.7 vs. 49.1 ± 2.6%). Onset of contractile response did not differ for 5HT (P = 0.26) and ERV (P = 0.93), but onset of ME5HT contraction was not initiated (P < 0.05) in HE steers until 10-4 compared with 10-5 M in LE-grazing steers. For Exp. 2, maximal contractile intensities achieved with DOI were 35% less (P < 0.05), whereas those achieved with 5CT were 37% greater (P < 0.05), in steers grazing HE pastures. Contractile response to CGS did not differ between pasture groups, and there was an absence of contractile response to BW7 in both groups. There were no differences between endophyte content in contractile responses after animals were finished for >91 d. Experiment 1 demonstrated that grazing of HE pastures for 89 to 105 d induces functional alterations in blood vessels, as evidenced by reduced contractile capacity and altered serotonergic receptor activity. Experiment 2 demonstrated that grazing HE pastures alters vascular responses, which may be mediated through altered serotonin receptor activities, and these alterations may be ameliorated by the removal of ergot alkaloid exposure as demonstrated by the absence of differences in finished steers. [ABSTRACT FROM AUTHOR]

L-Glutamate (Glu) is a major excitatory neurotransmitter responsible for neurotransmission in the vertebrate central nervous system. Vesicular Glu transporters VGLUT1 and VGLUT2 concentrate (50 mM) Glu [Michaelis constant (measuring affinity), or Km, = 1 to 4 mM] into synaptic vesicles (SV) for subsequent release into the synaptic cleft of glutamatergic neurons. Vesicular Glu transporter activity is dependent on vacuolar H+ -ATPase function. Previous research has shown that ergopeptines contained in endophyte-infected tall fescue interact with dopaminergic and serotoninergic receptors, thereby affecting physiology regulated by these neuron types. To test the hypothesis that ergopeptine alkaloids inhibit VGLUT activity of bovine cerebral SV, SV were isolated from cerebral tissue of Angus-cross steers that were naive to ergot alkaloids. Immunoblot analysis validated the enrichment of VGLUT1, VGLUT2, synaptophysin 1, and vacuolar H+ -ATPase in purified SV. Glutamate uptake assays demonstrated the dependence of SV VGLUT-like activity on the presence of ATP, H+ -gradients, and H+ -ATPase function. The effect of ergopeptines on VGLUT activity was evaluated by ANOVA. Inhibitory competition (IC50) experiments revealed that VGLUT-mediated Glu uptake (n = 9) was inhibited by ergopeptine alkaloids: bromocriptine (2.83 ± 0.59 µM) < ergotamine (20.5 ± 2.77 µM) < ergocornine (114 ± 23.1 µM) < ergovaline (137 ± 6.55 µM). Subsequent ergovaline kinetic inhibition analysis (n = 9; Glu = 0.05, 0.10, 0.50, 1, 2, 4, 5 mM) demonstrated no change in apparent Km. However, the maximum reaction rate (Vmax) of Glu uptake was decreased when evaluated in the presence of 50, 100, and 200 µM ergovaline, suggesting that ergovaline inhibited SV VGLUT activity through a noncompetitive mechanism. The findings of this study suggest cattle with fescue toxicosis may have a decreased glutamatergic neurotransmission capacity due to consumption of ergopeptine alkaloids. [ABSTRACT FROM AUTHOR]

To determine the effect of metabolic acidosis on expression of L-Gln, b-Gill, and L-Asp metabolizing enzymes and transporters, the relative content of mRNA, protein, or mRNA and protein, of 6 enzymes and 5 transporters was determined by real-time reverse transcription-PCR and immunobiot analyses in homogenates of kidney, skeletal muscle, and liver of growing lambs fed a common diet supplemented with canola meal (control; n = 5) or HC1-treated canola meal (acidosis; n = 5). Acidotic sheep had a 790% greater (P = 0.050) expression of renal Na+-coupled neutral AA transporter 3 mRNA and a decreased expression of renal glutamine synthetase mRNA (47% reduction, P = 0.037) and protein (57% reduction, P = 0.015) than control sheep. No change in renal cytosolic phosphoenolpyruvate carboxykinase (protein and mRNA), glutaminase (mRNA), or L-Glu dehydrogenase (protein) was found. In skeletal muscle, acidotic sheep had 101% more (P = 0.026) aspartate transaminase protein than did control sheep, whereas no change in the content of 3 Na+-coupled neutral AA transporters (mRNA) or 2 high-affinity L-Glu transporter proteins was found. In liver, no change in the content of any assessed enzyme or transporter was found. Collectively, these findings suggest that tissue-level responses of sheep to metabolic acidosis are different than for nonruminants. More specifically, these results indicate the potential capacity for metabolism of L-Asp and L-Glu by skeletal muscle, and L-Gln absorption by kidneys, but no change in hepatic expression of L-Gln metabolism, elaborates previous metabolic studies by revealing molecular-level responses to metabolic acidosis in sheep. The reader is cautioned that the metabolic acidosis model employed in this study differs from the increased plasma lactateinduced metabolic acidosis commonly observed in ruminants fed a highly fermentable grain diet. [ABSTRACT FROM AUTHOR]

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 ≈ 260 kg) were assigned to water (control), ruminal cornstarch (partially hydrolyzed by α-amylase; SH), or abomasal Sil infusion treatments (n = 6) and fed an alfalfa-cube-based diet at 1.3 x NEm 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 re- verse 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 < 0.02) by jejunal and duodenal than by ileal epithelia. The content of GLUT5 mRNA in small intestinal epithelia was not affected (P > 0.16) by either SH infusion treatment. In contrast, GLUT2 and SGLT1 mRNA content in the ileal epithelium was increased (P < 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 GLTUT2 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 epitheha, assuming that altered SugT mRNA contents reflect the altered transport functional capacities. [ABSTRACT FROM AUTHOR]

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 (ATB0,+>), b0,+> (b0,+> 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) glucose-derived energy (from starch hydrolysate, SH), 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 ATB0,+> mRNA greater than jejunal or ileal expression; ileal expression of b0,+>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. [ABSTRACT FROM AUTHOR]

It is well established that grazing Neotyphodium coenophialum-infected forages results in reduced BW gain and serum prolactin concentrations of cattle. The objective of this study was to determine the potential effects of toxic endophyte-infected tall fescue consumption on blood metabolites, carcass characteristics, and content of proteins critical for AA metabolism in the liver, kidney, and LM tissue of growing steers. Steers grazed a low toxic endophyte (LE; 0.023 µg/g ergot alkaloids) tall fescue-mixed grass pasture (n = 9; BW = 266 ± 10.9 kg; 5.7 ha) or a high toxic endophyte (HE; 0.746 µg/g of ergot alkaloids) tall fescue pasture (n = 10; BW = 267 ± 14.5 kg; 5.7 ha) from June 14 through at least September 11 (≥89 d). No difference was observed for BW (P < 0.10) for the overall 85-d growth period. Also, no differences were observed for ribeye area/100 kg of HCW (P> 0.91), backfat (P> 0.95), or backfat/100 kg of HCW (P> 0.67). However, ADG (P < 0.01), final BW (P < 0.05), HCW (P < 0.01), dressing percentage (P < 0.01), ribeye area (P < 0.01), whole liver wet weight (P < 0.01), and whole liver wet weight/100kg of end BW (P < 0.01) were greater for LE steers than HE steers. After 85 d of grazing, serum concentrations of alkaline phosphatase (P < 0.05), alanine aminotransferase (P < 0.01), aspartate arninotransferase (P < 0.03), cholesterol (P < 0.01), lactate dehydrogenase (P < 0.01), and prolactin (P < 0.01) were less for HE than LE steers. At slaughter, hepatic content of cytosolic phosphoenolpyruvate carboxykinase (P < 0.01) was greater in HE steers than LE steers. Hepatic content of aspartate aminotransferase (P < 0.01) also was greater, whereas renal and LM content were not (P ≥ 0.42). No differences (P ≥ 0.15) were observed for hepatic, renal, and LM content of alanine aminotransferase, glutamate dehydrogenase, glutamine synthetase, and 3 glutamate transport proteins. These data indicate that the HE steers displayed classic endophyte toxicity symptoms for growth arid blood variables, classic symptoms that were concomitant with novelly identified altered glucogenic capacity of the liver and decreases in carcass characteristics. [ABSTRACT FROM AUTHOR]

In ruminants microbial-derived nucleic acids are a major source of X 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 alfafa cube-based die at 1.33 x NEm 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, or ruminal water infusion/abomasal water 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). [ABSTRACT FROM AUTHOR]

Grazing -infected forages results in a variety of reduced animal performance parameters, collectively known as "fescue toxicosis." The initial, limited evaluations of hepatic mechanisms affected by fescue toxicosis have used transcriptomic expression profiling of experimental phenotypes developed by short-term feeding of concentrated ergot alkaloids or fescue seeds to rodents and steers. To assess the effects of fescue toxicosis in growing cattle using a commercially relevant phenotype, we induced fescue toxicosis in beef steers by summer-long grazing (89 to 105 d) of a single high toxic endophyte-infected tall fescue pasture (HE; 0.746 μg/g ergot alkaloids; 5.7 ha; = 10; BW = 267 ± 14.5 kg) vs. a low toxic endophyte tall fescue-mixed pasture (LE; 0.023 μg/g ergot alkaloids; 5.7 ha; = 9; BW = 266 ± 10.9 kg). High toxic endophyte tall fescue-mixed pasture steers had decreased BW (313 vs. 338 kg) and an increased potential for hepatic gluconeogenesis from AA-derived carbons. To gain a greater perspective into fescue toxicosis-induced hepatic metabolism and identify candidate regulatory mechanisms, the goal of the current research was to examine liver samples for changes in gene (mRNA) expression profiles using a Bovine Affymetrix microarray and selected reverse-transcription PCR and immunoblot analyses. The expression (false discovery rate < 10%; < 0.01) of 147 genes was increased (7 to 268%) and that of 227 was decreased (4 to 87%) in livers of HE vs. LE steers. The top (1) functional gene category was cell-mediated immune response (33 genes; ≤ 0.012), (2) canonical cell signaling pathway was primary immunodeficiency signaling (8 genes; ≤ 0.0003), and (3) canonical metabolic pathways were oxidative phosphorylation (5 genes; ≤ 0.016) and purine metabolism (8 genes; ≤ 0.029). High toxic endophyte tall fescue-mixed pasture steers had increased ( ≤ 0.022) expression of genes critical for increased (1) Pro () and Ser () synthesis, (2) shunting of AA carbons into pyruvate () and ATP synthesis (, , , COX4, , and ), and (3) mitochondrial mass (COX4). Targeted reverse-transcribed PCR or immunoblot assays corroborated ( ≤ 0.035) these latter microarray findings for , , , , and COX4. Moreover, network analysis identified glucocorticoid receptor-mediated signaling as the most probable mechanism to coordinate the above findings. These results greatly extend our knowledge of the consequences of summer-long grazing of endophyte-infected tall fescue to the hepatic metabolism of growing steers.

Selenium (Se) is an important trace mineral that, due to deficiencies in the soil in many parts of the USA, must be supplemented directly to the diet of foraging cattle. Both organic and inorganic forms of dietary Se supplements are available and commonly used, and it is known that Se form affects tissue assimilation, bioavailability, and physiological responses. However, little is known about the effects of form of dietary Se supplements on gene expression profiles, which ostensibly account for Se form-dependent physiological processes. To determine if hepatic transcriptomes of growing beef (Angus-cross) heifers (0.5 kg gain/day) was altered by form of dietary supplemental Se, none (Control), or 3 mg Se/day as inorganic Se (ISe, sodium selenite), organic (OSe, Sel-Plex®), or a blend of ISe and OSe (1.5 mg:1.5 mg, Mix) Se was fed for 168 days, and the RNA expression profiles from biopsied liver tissues was compared by microarray analysis. The relative abundance of 139 RNA transcripts was affected by Se treatment, with 86 of these with complete gene annotations. Statistical and bioinformatic analysis of the annotated RNA transcripts revealed clear differences among the four Se treatment groups in their hepatic expression profiles, including (1) solely and commonly affected transcripts; (2) Control and OSe profiles being more similar than Mix and ISe treatments; (3) distinct OSe-, Mix-, and ISe-Se treatment-induced "phenotypes" that possessed both common and unique predicted physiological capacities; and (4) expression of three microRNAs were uniquely sensitive to OSe, ISe, or Mix treatments, including increased capacity for redox potential induced by OSe and Mix Se treatments resulting from decreased expression of MiR2300b messenger RNA. These findings indicate that the form of supplemental dietary Se consumed by cattle will affect the composition of liver transcriptomes resulting, presumably, in different physiological capacities.

Forty-eight Angus x Hereford steers (initial BW = 336 +/- 8.3 kg) were used in a 56-d study to evaluate growth and endocrine responses to continuous or discontinuous grazing of high-endophyte-infected Kentucky-31 (K; > 57% infestation rate) or low-endophyte-infected Johnstone tall fescue (J; < 1% infestation rate) and implantation with 0 or 24 mg/steer of estradiol-17 beta (E2; Compudose). Steers were allotted by weight to eight 3-ha paddocks (four paddocks of each fescue variety) with six steers per paddock. Two paddocks of each variety were grazed continuously (KK and JJ), whereas steers on the remaining four paddocks were rotated every 14 d from K to J (KJ) or from J to K (JK). Three steers in each paddock were implanted with E2 on d 0. The study extended from May 25, 1988 to July 20, 1988 with steers exposed to potential heat-stress conditions for 52 d of the 56-d study. Body weights were obtained on d 0, 28, and 56, and blood samples were taken on d 28 and 56. Overall ADG, serum prolactin, and serum alkaline phosphatase activity were greater (P < .05) in JJ than in KK steers. Rotation from K to J did not increase overall ADG, prolactin, insulin-like growth factor I (IGF-I), or alkaline phosphatase activity compared with the continuously grazed KK, whereas JK steers had lower (P < .10) ADG, prolactin, and alkaline phosphatase activity than JJ steers. Estradiol-17 beta increased (P < .10) IGF-I in JJ, KJ, and JK steers but not in KK steers.(ABSTRACT TRUNCATED AT 250 WORDS)

We tested the hypothesis that the increase in LH secretion associated with elevated feed intake in nutritionally growth-restricted lambs is due to increased availability of tyrosine (TYR). Ovariectomized ewe lambs with abomasal cannulae were fed a complete diet in amounts (0.6 Mcal of net energy/day) sufficient to maintain a body weight of 21.7 +/- 1.0 kg between 10 and 32 wk of age. At 32 wk of age, lambs were assigned at random to one of three treatments: 1) maintenance feeding + abomasal infusion of water (M; n = 6); 2) maintenance feeding + abomasal infusion of TYR (M + TYR; n = 7); or 3) twice maintenance feeding + abomasal infusion of water (TM; n = 6). TYR (1.25 g/100 ml water) or water (100 ml) was delivered as a bolus injection at 0600, 1200, 1800, and 2400 h daily for 23 days. Plasma concentrations of TYR 10 and 22 days after initiation of treatments were higher (p less than 0.001) in the TM and M + TYR groups compared to the M group and were elevated (p less than 0.001) in the M + TYR group compared to the TM group. Concentrations of TYR in hypothalami were higher (p less than 0.01) in the M + TYR and TM groups than in the M group, and greater (p less than 0.005) in the M + TYR group compared to the TM group. Overall, there was a linear (p less than 0.001) correlation between plasma and hypothalamic concentrations of TYR.(ABSTRACT TRUNCATED AT 250 WORDS)

Ten Hampshire x Western wether lambs (means weight = 30.1 kg) equipped with ruminal and abomasal cannulas were fed either low-Acremonium coenophialum (AC) Kentucky-31 (less than 1% infected) or high-AC G1-307 (greater than 95% infected) varieties of tall-fescue (TF) hay of similar nutrient composition in a completely randomized design. Lambs were housed in metabolism crates at 21 +/- 1 degrees C and fed 552 g DM/d of ground hay at 0800 and 2000. A 10-d adaptation preceded 7 d of sample collection. Levels of water and DM voluntarily consumed by the low-AC group during the adjustment period were held constant for both treatment groups throughout the collection period by intraruminal insertion of unconsumed DM and water. Fixed water intake markedly reduced voluntary water intake but it alleviated previous depressions in voluntary DM intake in lambs fed high-AC. Mean daily respiration and heart rates, rectal temperature and hematocrit were not affected (P greater than .10) by treatment. Compared with high-AC, lambs fed low-AC retained a greater (P less than .10) amount of N (1.8 vs 1.1 g/d) and a greater (P less than .10) percentage of their N intake (16.4 vs 9.9%). Abomasal total and bacterial N flow and ruminal digestion of cell wall components were not affected (P greater than .10) by treatment. Total tract digestion of DM, NDF and ADF was lower (P less than .01) for high- than for low-AC. Serum prolactin concentration was higher (P less than .10) for lambs fed low- than for those fed high-AC TF, but serum cortisol and thyroxine concentrations were not affected (P greater than .10) by treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

A 100-d growth study was conducted to evaluate performance and plasma amino acid (AA) responses of 96 crossbred beef calves (220 kg) with ad libitum access to corn silage and supplemented with ground soybeans (GSB) with or without added fishmeal (FM) and (or) rumen-protected lysine (Lys). Calves were allotted by breed, sex and weight to four treatments with three replicate pens of eight calves per pen. The treatments were: GSB, GSB + Lys, GSB + FM and GSB + FM + Lys. The isonitrogenous supplements were top-dressed on corn silage once daily at a level of 2.27 kg/hd, with FM providing one-half of the supplemental N in FM-containing supplements. The Lys-containing supplements provided a daily intake of 6.0 g/hd of rumen-protected Lys. Dry matter intake was similar (P less than .10) for all treatments. Overall ADG and feed efficiency of GSB calves averaged .83 kg/d and 7.39 kg feed/kg gain, respectively, and were 14% lower than the mean of calves fed supplements containing FM and(or) Lys. Lysine was not the principal factor limiting growth because the inclusion of Lys alone in the GSB-containing supplements did not improve (P greater than .10) ADG, feed efficiency or plasma AA concentrations. In contrast, FM supplementation increased (P less than .05) ADG, feed efficiency and plasma concentrations of total AA, total essential AA and total nonessential AA. The inclusion of Lys in the GSB + FM-containing supplement resulted in no further improvement (P greater than .10) in performance.(ABSTRACT TRUNCATED AT 250 WORDS)

Two experiments were conducted to compare Mg bioavailability from Mg oxide (MgO) vs Mg hydroxide (Mg(OH)2) fed in either a completely mixed diet or a mineral supplement. In Exp. 1, these Mg sources were incorporated into completely mixed diets and offered to 15 steers (282 kg) allotted to three treatments: control diet containing .19% Mg, control plus .2% added Mg as MgO, or control plus .2% added Mg as Mg(OH)2. Each calf was fed 5 kg/d of the respective diet during 10-d adjustment and 7-d collection periods. Blood samples were collected on d 1, 3 and 7. Mg supplementation increased (P less than .01) fecal and urinary Mg excretions, whereas apparent Mg absorption (%) and retention were similar (P greater than .10) for all treatments. Plasma Mg concentrations were similar (P less than .10) for calves supplemented with MgO and Mg(OH)2 but were higher (P less than .05) for Mg supplemented than for control calves on d 7. In Exp. 2, these Mg sources were incorporated into mineral supplements and offered free choice to 30 spring-calving beef cows gazing tetany-inducing pastures from March 6 to May 1. Each of three groups of 10 cows was assigned to a 5.7-ha tall fescue pasture and offered either a control supplement or a supplement containing 40% MgO or Mg(OH)2. Blood samplers were collected on d 0, 7, 14, 28, 42 and 56. Plasma Mg concentrations were not different (P greater than .10) for cows offered MgO and Mg(OH)2 but were higher (P less than .01) for Mg-supplemented than for control cows on d 28, 42 and 56.(ABSTRACT TRUNCATED AT 250 WORDS)

Experiments were conducted with rats and mice to evaluate the effect of the consumption of endophyte (Acremonium coenophialum) and associated toxin(s) infected tall fescue on humoral and cellular aspects of immune function. Treatment diets were: (1) rodent chow (RC) or (2) rodent chow mixed 1:1 (w/w) with endophyte infected (E+) or (3) non-infected (E-) tall fescue seed. Rats fed the E+ diet in experiment 1 (43 days) exhibited a lower (P less than 0.05) serum titer to sheep red blood cell (SRBC) immunization than those fed the E- diet (38.4 vs 131.3). The E+ rats also had lower (P less than 0.01) white cell counts than either RC or E- groups (5225 vs 8959 and 7491/mm3). Spleen cells from mice fed the E+ diet for 37 days exhibited a reduced (P less than 0.05) response to the mitogens Concanavalin A and lipopolysaccharide. Flow cytometric analysis revealed a significant (P less than 0.01) 42% increase in T suppressor cell numbers in spleens of mice fed the E+ vs RC diets.

Effects of two microbial feed supplements on microbial activities in rumen-stimulating cultures and the rumens of steers fed a fescue hay-based roughage diet were evaluated. The yeast culture supplement contained Saccharomyces cerevisiae (1.4 to 4.2 x 10(9) colony-forming units [cfu]/g), whereas the mixed microbial supplement contained yeast, lactobacilli and enterococci (1.4 to 2.7 x 10(9) cfu/g, 1.2 to 2.3 x 10(9) cfu/g, and 1.5 to 2.6 x 10(10) cfu/g, respectively). Concentrations of viable yeast cells were increased consistently in continuous cultures and rumens of steers receiving either supplement (1 g/kg of feed). However, neither supplement consistently altered the relative concentrations of volatile fatty acids or ammonia in continuous cultures and rumens of steers. The pH tended to be greater (P = .13) in continuous cultures receiving yeast culture supplement than in cultures receiving the unsupplemented diet (6.50 vs 6.36), but pH in the rumens of steers was not affected by the supplements. Concentrations of cellulolytic microorganisms in cultures and the rumens of steers receiving supplements containing only yeast were from 5 to 40 times greater than those observed in cultures or steers receiving the unsupplemented diet. Supplements that had been treated with heat (121 degrees C for 15 min) to inactive yeast cells did not alter the concentrations of cellulolytic bacteria in rumen-stimulating cultures. These results suggest that live yeast culture supplements stimulate growth of cellulolytic microorganisms in the rumen.

Twelve Angus x Hereford heifer calves (233 kg) were fitted with abomasal infusion cannulas and used to study N and endocrine responses to abomasally infused arginine (Arg). Heifers were allotted randomly to three treatment groups and received continuous abomasal infusions (2 liters/d) of water (CON) or Arg solutions providing .33 g Arg.HCl/kg BW (LOW) or .50 g Arg.HCl/kg BW (HIGH) each day. A 12-d dietary adjustment period preceded a 7-d infusion and collection period. Each calf received 4,544 g DM/d of a basal diet in equal portions at 0600, 1200, 1800 and 2400. Calves were housed in individual metabolism crates and fitted with urinary bladder catheters for total excreta collection. On d 1 and 5, blood samples were collected at 15-min intervals for 8 h between 1200 and 2000. Single samples were obtained at 1400 on remaining days. The infusion of Arg increased the quantity of N retained by heifers (P less than .01) and the percentage of total N retained (P less than .10); however, no differences were observed between LOW and HIGH heifers. Increased (P less than .01) urinary N excretion by Arg heifers was associated with greater (P less than .05) quantities of urinary urea N and ammonia N. Blood urea N and serum Arg concentrations were highest (P less than .05) in Arg heifers, whereas total serum AA concentrations were lower (P less than .05) in Arg heifers than in CON heifers. Serum glucose and insulin concentrations were not affected (P greater than .10) by treatment. Characterization of somatotropin (STH) profiles revealed that amplitude and frequency of STH pulses were not affected (P greater than .10) by treatment, whereas mean (P less than .10) and basal (P less than .05) STH concentrations were elevated in HIGH compared to LOW heifers on d 1 and 5. The similar N retention responses of LOW and HIGH heifers and similar STH profiles of CON and LOW heifers suggest that the stimulatory effect of the HIGH dose on STH secretion occurred only after tissue N requirements had been satisfied.

The hypothesis that prepubertal ewe lambs are metabolically different from postpubertal ewes was tested. Ovariectomized ewes (4 years of age; n = 4) and lambs (6 months of age; n = 4) were fasted for 72 hr. Serum concentrations of insulin, glucose, urea nitrogen, and free fatty acids (FFA) were measured in blood samples taken at 6-hr intervals between 30 hr before and 72 hr after feed removal. Serum concentrations of urea nitrogen and glucose were not different (P greater than 0.20) between age groups before fasting. Serum concentrations of insulin in ewes increased toward the end of the prefast period whereas those in lambs did not (age x time, P less than 0.01). Serum concentrations of FFA in ewes tended to be lower (P less than 0.07) than those in lambs prior to fasting. During fasting, concentrations of insulin decreased (P less than 0.02) over time in ewes and lambs and did so in a similar manner (age x time, P greater than 0.70). Urea nitrogen increased (P less than 0.0001) in both fasted ewes and fasted lambs in a comparable manner (age x time, P greater than 0.20). Concentrations of glucose during fasting were not significantly affected (P greater than 0.90) by age. There was a tendency (P = 0.08) for concentrations of glucose to change over time but the pattern did not appear to be related to fasting. During fasting, concentrations of FFA tended to be higher (P less than 0.07) in lambs than in ewes and increased (P less than 0.0001) in both groups in a similar fashion (age x time, P greater than 0.10). The findings herein suggest that turnover of FFA in lambs may be slightly greater than that in ewes during the fed and fasted states.

The effects of FFA on circulating LH and GH concentrations in ovariectomized ewe lambs were investigated. Lambs (n = 14) were weaned at 2.5 months, ovariectomized at 6.5 months, and used at 8.5 months of age. From weaning until day 0 of the experiment, lambs were fed to maintain body weights (23 kg). On day 0, serum FFA concentrations and mean serum LH concentrations and number and amplitude of LH pulses, as assessed in blood samples collected every 12 min for 4 h, were 6.4 +/- 0.6 mg/100 ml, 0.57 +/- 0.08 ng/ml, 0.45 +/- 0.09 pulses/h, and 0.73 +/- 0.11 ng/ml, respectively. Double the maintenance feeding, beginning day 1, increased (P less than 0.01) body weights by 16% and LH pulse frequency by 82%, but had no effect (P greater than 0.1) on FFA concentrations, mean LH concentrations, or LH pulse amplitude by day 14. On day 14, lambs were infused with lipid (n = 9; 95.8 mg/min) or 0.9% saline solution (n = 5) for 8 h. Blood samples were collected at 12-min intervals for 12 h, beginning 4 h before infusions. FFA levels increased (P less than 0.01) in lipid-infused animals to 27.6 +/- 2.9 mg/100 ml by 4 h of infusion. Mean LH concentrations and LH pulse frequency and amplitude were unaffected (P greater than 0.1) by treatment. In contrast, mean GH concentrations and GH pulse frequency, which were similar (P greater than 0.1) between groups before infusion (14.0 +/- 0.8 ng/ml and 0.36 +/- 0.07 pulses/h, respectively) were decreased by FFA treatment by 51% (P less than 0.01) and 81% (P less than 0.006), respectively. GH pulse amplitude was highly variable and unaffected (P greater than 0.1) by treatment. In summary, elevated FFA levels appear to inhibit the release of GH, but not LH, in the ovariectomized ewe lamb.

Twelve growing Angus heifers averaging 282 kg in weight were fed a tall fescue hay-based diet in a completely randomized design experiment to determine sulfur (S) and nitrogen (N) metabolism and bioavailability of different forms of supplemental S:elemental S (E), sodium sulfate (SU) and DL-methionine (M). The study included a 72-h adaptation phase that was followed immediately by a 5-d retention phase. Control (C) heifers were fed a ground fescue hay diet twice daily throughout the study. Supplementation with .15% S from E, SU or M to the basal diet began at h 0. Differences in total blood S, plasma S and ruminal NH3 N were dependent on sampling time in the 72-h adaptation phase. Urinary S excretion during 0 to 72 h was greater for S-supplemented calves than for C. Mean molar proportions of acetate, butyrate, isobutyrate and valerate were affected by form of S supplementation. No treatment differences (P greater than .10) were noted in total VFA concentrations. Sulfur supplementation increased daily urinary sulfate (P less than .01), total urinary S (P less than .01) and fecal S (P less than .01) excretion during the retention phase. Methionine and SU heifers had the greatest urinary S losses, and fecal S was highest for E (P less than .01). No differences (P greater than .10) in DM digestion occurred among treatments. N retention (g/d) and N retention as a percentage of intake averaged 5.9, 8.5; 13.2, 18.7; 13.8, 20.5; and 11.3, 19.2, respectively, for C, E, SU and M. Sulfur retention (g/d) was increased (P less than .10) by S supplementation. Sulfur supplementation of a basal fescue hay diet increased S retention transitorily, although much of the added S was lost through excretory routes. Supplementing S as E, SU or M appeared to be equally beneficial, despite differences in route and extent of S excretion among the three S sources.

Sixteen wether lambs (25 kg) were fitted with abomasal infusion cannulas and used to study N and endocrine responses to abomasal infusions of arginine (ARG) or ornithine (ORN). Lambs were randomly allotted to four treatment groups and abomasally infused with solutions of water (CON), ARG, ORN or UREA. The ARG solution provided .50 g ARG.HCl/kg BW and was equimolar with ORN.HCl (.40 g/kg). UREA (.28 g/kg) was isonitrogenous with ARG and served as a positive N control. Lambs were housed in metabolism crates for excreta collection and received 729 g DM/d of a 13.7% CP diet in equal portions four times daily. Following a 7-d dietary adjustment period, lambs were infused continuously (2 liters/d) with water for a 5-d preliminary collection period (Period 1), which immediately preceded a 7-d infusion and collection period (Period 2). Sequential blood samples were taken at 15-min intervals for 8 h between 1200 and 2000 on d 4 of both periods. Single samples were obtained at 1500 on remaining days. Nonrepeated measurements were analyzed as a completely randomized design, whereas repeated measurements were analyzed as a split-plot over time. Period 2 measurements were adjusted using covariance techniques if differences among treatment groups were observed for Period 1. Contrasts used in determining treatment effects were: CON vs UREA, CON vs ARG + ORN, and ARG vs ORN. Nitrogen retention was similar for all treatment, suggesting that dietary N was not limiting. Arginine and ORN increased serum ornithine (P less than .05), blood urea N (BUN; P less than .10) and urinary urea N excretion (P less than .01), whereas ARG increased (P less than .05) serum arginine and UREA increased (P less than .01) BUN and urinary urea N. Serum insulin and glucose were not affected by treatment. Compared with CON, ARG and ORN increased (P less than .05) mean somatotropin (STH) concentration (13.8 vs 16.9 and 18.4 ng/ml) and amplitude of STH pulses (9.8 vs 15.1 and 17.8 ng/ml), whereas CON and UREA were similar. Abomasal infusions of ARG and ORN were equally efficacious in stimulating ovine STH secretion when dietary N intake was not limiting.

Postruminal utilization of methionine by growing steer calves fed ground shelled corn, urea, and cottonseed hulls adequate in sulfur was studied by abomasal infusion of graded quantities of methionine with lysine adequate. A 6 X 6 Latin square of treatments was replicated in which infusions provided lysine and methionine in grams per day: A) 0,0; B) 24,0; C) 24,4; D) 24,8; E) 24,12; and F) 24,12 plus 140 g sodium caseinate. Treatments were brought to volume in 2 liters of water and infused continuously over 24 h. Responses measured were nitrogen retention, urea nitrogen in plasma, and concentrations of free amino acids. Retention of nitrogen was increased in steers abomasally infused with lysine or lysine in combination with graded quantities of methionine. Infusion of 4, 8, or 12 g/day of methionine with constant lysine (24 g/day) did not alter nitrogen retained from lysine infused alone. Infusion of a lysine-methionine-sodium caseinate positive control raised nitrogen retention compared with no caseinate. Lysine of plasma was higher in treatments in which lysine was infused compared with the negative control. Methionine increased linearly with infusion of incremental quantities. Methionine was not limiting when infused prostruminally with adequate lysine to growing steers fed the urea-supplemented diet with sulfur adequate.

Eight Angus heifer calves (234 kg) were assigned to either a high (HP; 126 g N/d) or low (LP; 66.5 g N/d) protein intake. Calves received 4.8 kg DM/d consisting of 30% cottonseed hulls and 70% corn-soybean meal in equal portions at 4-h intervals. Single doses of 14C- and 15N-urea and 15N-ammonium sulfate were injected into the blood urea-N (BUN) and ruminal NH3-N (RAN) pools, respectively, to measure rate of flux through, and transfer of N between, these and bacterial N. Nitrogen balance was greater (P less than .05) for HP than for LP (56.9 vs 25.1 g N/d), but abomasal N flow as a percentage of N intake was greater (P less than .05) for LP than for HP (124 vs 71.1%). Pool size and net synthesis rate for both RAN and BUN pools were greater (P less than .05) for HP than for LP. Calves fed HP degraded more (P less than .05) BUN in the gastrointestinal tract than calves fed LP (37.4 vs 14.0 g N/d). Quantities of RAN absorbed from the rumen also were greater (P less than .05) for HP than for LP (14.2 vs 2.8 g N/d). The proportion of total gastrointestinal BUN degradation occurring in the rumen averaged 53 and 26% for LP and HP. Data are interpreted to suggest that net incorporation of BUN into bacterial protein (urea recycling) is inversely related to level of protein intake.

Two completely randomized design experiments were conducted, using either 10, 38-kg (Exp. 1) or 10, 26-kg (Exp. 2) Hampshire x Western wether lambs, to study the effects of alpha-ketoglutarate (AKG) administration on N metabolism. Lambs were fed 890 and 885 g DM/d in Exp. 1 and 2, respectively, of corn-cottonseed hull basal diets with urea added to attain CP levels of 10.6 and 10.5% in Exp. 1 and 2, respectively. Experiments consisted of 10 d of adaptation followed by 7 d of infusion and excreta collection. Lambs were infused continuously either i.v. (Exp. 1) or abomasally (Exp. 2) with control solutions (CON) or solutions containing 41.3 g AKG/d (AKG). In Exp. 1, fecal and urinary N excretion and N retention were not affected (P greater than .10) by treatment. Compared to CON in Exp. 2, AKG infusion increased (P less than .10) fecal N output (6.6 vs 5.9 g N/d) but did not affect (P greater than .10) the amount of N retained (4.4 vs 3.5 g N/d). Compared to CON, AKG increased (P less than .10) urinary NH3 N excretion in Exp. 1. Serum urea N was lower (P greater than .10) for AKG than for CON in Exp. 1 but was not affected (P greater than .10) by treatment in Exp. 2. In Exp. 1, AKG appeared to reduce activities of several serum enzymes that function in amino N metabolism. In Exp. 1, compared to CON, AKG decreased (P less than .10) aspartate but increased (P less than .10) asparagine in serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Thirty pregnant beef cows were utilized to determine the effects of winter Mg supplementation regimen on blood minerals after cows were turned to a spring tetanigenic tall fescue pasture. The winter Mg treatments were (1) tall fescue hay with free access to a Mg-deficient salt-mineral mix, (2) 6.4 kg of corn silage dry matter plus 114 g MgO/d and (3) tall fescue hay with free access to a salt-mineral mix containing 40% MgO from January 1 to February 15. All cows were then placed on the same tetanigenic pasture from February 15 to April 17 with free access to a Mg-deficient salt-mineral supplement. Forage Mg remained below .2% during the experiment. Forage Ca, P and Al changed throughout the spring, attaining maximum values of .35%, .46% and 415 ppm, respectively. Forage N and K also increased throughout the spring, reaching values of 3.5 and 3.8%, respectively, at the April 3 sampling. The forage K:(Ca + Mg) ratio approached 2.2 by March 26, which coincided closely with the average tetany date (March 29). Serum Mg averaged 1.97, 3.58 and 2.06 mg/dl for treatments 1, 2 and 3, respectively, on February 15 before turning cows to pasture. There were no treatment differences for serum Ca, P and K during the experiment. Eight cows exhibited symptoms of grass tetany (collapse) on an average date of March 29. Winter Mg supplementation provided little long-term protection against hypomagnesemia after turning cows to tetanigenic pasture, indicating that cows must have a supplemental source of Mg during this critical period.