Bovine neuronal vesicular glutamate transporter activity is inhibited by ergovaline and other ergopeptines.

l-Glutamate (Glu) is a major excitatory neurotransmitter responsible for neurotransmission in the vertebrate central nervous system. Vesicular Glu transporters VGLUT1 and VGLUT2 concentrate (50mM) Glu [Michaelis constant (measuring affinity), or K(m),=1 to 4mM] 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 (IC(50)) experiments revealed that VGLUT-mediated Glu uptake (n=9) was inhibited by ergopeptine alkaloids: bromocriptine (2.83±0.59µM)

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.

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).

Gene expression profiling indicates an increased capacity for proline, serine, and ATP synthesis and mitochondrial mass by the liver of steers grazing high vs. low endophyte-infected tall fescue.

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.

Glutamine synthetase and alanine transaminase expression are decreased in livers of aged vs. young beef cows and GS can be upregulated by 17ß-estradiol implants.

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 (17ß-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 ( ≥ 0.17). Hepatic ALT (78% and 61%) and GS (52% and 71%) protein content (Exp. 1 and 2, respectively) was decreased ( ≤ 0.01), whereas GTRAP3-18 (an inhibitor of EAAC1 activity) increased ( ≤ 0.01; 170% and 136%) and AST, GLT-1, and EAAC1 contents did not differ ( ≥ 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 ( = 0.10) free Gln (15.0%) than young cows, whereas other AA concentrations did not differ ( 0.26). In Exp. 3, 14 aged (> 10 yr) cows were randomly allotted ( = 7) to sham or COMPUDOSE (25.7 mg of 17ß-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 ( ≥ 0.96). Implant increased ( ≤ 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 ( ≥ 0.16). We conclude that hepatic expression of ALT and GS are reduced in aged vs. young cows, and administration of 17ß-estradiol to aged cows increases plasma estradiol and hepatic GS, but not that of other proteins that support hepatic Glu metabolism.

Free fatty acids suppress growth hormone, but not luteinizing hormone, secretion in sheep.

An experiment was conducted to determine the effects of exogenously administered FFA on GH and LH secretion in sheep. Ovariectomized ewes received iv infusions of a mixture of FFA (166 mg/min; n = 5) or 0.9% saline (n = 4) for 10 h. Jugular blood was sampled every 15 min for 14 h, beginning 4 h before initiation of infusion. After 8 h of FFA or saline treatment, each ewe received a pituitary challenge of 10 micrograms GRF and 1 microgram GnRH, administered together as an iv bolus. Lipid infusion increased (P less than 0.01) serum FFA concentrations to levels characteristic of those in fasted sheep [23.0 +/- 0.8 mg/100 ml (mean +/- SE)]. Frequency of GH pulses (P less than 0.01) and the GH response to GRF (P less than 0.0001) were suppressed by FFA treatment. Mean serum GH concentrations increased gradually (P less than 0.01) during the 10-h infusion period in saline-treated but not lipid-treated, ewes. This finding may reflect diurnal changes in somatotrope secretory activity that are blocked by FFA. Mean serum LH concentrations, LH pulse frequency and amplitude, and the LH secretory response to GnRH were unaffected by FFA or saline infusion. In agreement with previous work in sheep and other species, these results provide evidence for an inhibitory effect of FFA on GH release. The exact mechanism responsible for this action, however, remains to be elucidated. Finally, acutely elevated FFA levels do not appear to influence LH secretion in the ovariectomized ewe.

Effects of free fatty acids on luteinizing hormone and growth hormone secretion in ovariectomized lambs.

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.

N-methyl-d, l-aspartate stimulates growth hormone but not luteinizing hormone secretion in the sheep.

The objective of this experiment was to determine the effects of N-methyl-d, l-aspartate (NMA) on luteinizing hormone (LH) and growth hormone (GH) secretion in castrated male sheep. Blood was sampled from Hampshire wethers every 15 min for 8 hr on day 1. At 4 and 6 hr after the initiation of the experiment, wethers were treated i.v. with NMA at a dose of 12 mg/kg body weight (n = 5) or .9% saline (n = 5). The dosage of NMA was within the range of doses that was previously demonstrated to stimulate LH secretion in monkeys. Blood samples were also collected every 15 min for 1 hr on day 2, beginning 24 hr after the first injection of NMA or saline. Treatment with NMA had no effect on mean LH concentrations, LH pulse frequency or LH pulse amplitude during the 4 hr period following the first injection on day 1. On day 2, however, mean LH concentrations were lower (p less than .01) in NMA versus saline-treated wethers. Conversely, administration of NMA evoked a dramatic increase (p less than .02) in mean GH concentrations on day 1. The mechanisms responsible for the effects of NMA described herein and whether or not these effects are relevant to the physiological control of LH and GH release in the sheep warrants further scrutiny.

The effect of the endophyte (Acremonium coenophialum) and associated toxin(s) of tall fescue on serum titer response to immunization and spleen cell flow cytometry analysis and response to mitogens.

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.

Nutrient utilization in lambs fed diets high in sodium or potassium.

Twenty-four wether lambs averaging 47 kg were used to study the effects of dietary K and Na additions on metabolism of lambs. Lambs were randomly allotted to four treatments and fed 900 g/d of the following: control diet consisting of 50% ground tall fescue hay and 50% concentrate; K diet calculated to contain 4% K with K added as KCl; Na diet calculated to contain 4% Na with Na added as NaCl, and K-Na diet containing 2% K and 2% Na with K and Na added from the same sources as in the K and Na diets. Water intake and urine excretion were highest for lambs fed the Na diet. Dry matter (DM) and crude protein (CP) digestibilities were similar among treatments. The K-Na diet resulted in decreased (P less than .05) acid detergent fiber (ADF) digestibility compared with lambs fed the Na diet. Nitrogen (N) retention was highest (P less than .05) in lambs fed the K diet and lowest in those fed the Na diet. Rumen NH3-N was lower (P less than .05) in lambs fed the Na diet compared with controls. Rumen K at 2 and 6 h postfeeding was highest (P less than .05) and rumen Na at 6 h lowest (P less than .05) for lambs fed the K diet. Plasma K values at both sampling times were highest (P less than .05) in lambs fed the K diet. At 6 h postfeeding, plasma Mg was higher in control lambs compared with those fed the K-Na diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Short-term metabolic response in wethers to high dietary protein intake prior to fasting.

Twenty crossbred wether lambs (average body weight 38.9 kg) were examined for selected metabolic responses during fasting as affected by dietary protein level prior to fasting. Animals were fed diet analyzed at 10.4 and 20.3% crude protein (diets A and B, respectively) for 3 weeks and then fasted for 7 days. jugular blood samples were collected 4, 28, 52, 76, 100, 124 and 148 hr after the last feeding. Plasma glucose declined (P less than .05) between 4 and 52 hr, after which no further differences in plasma glucose occurred within or between treatment groups. Plasma urea N (PUN) was higher (P less than .05) in lambs fed diet B at 4, 28, and 52 hr postfeeding. In wethers fed diet A, PUN increased (P less than .05) from 28 to 76 hr postfeeding and then declined to prefasting levels. In wethers fed diet B, PUN did not begin to decrease until 76 hr postfeeding and then gradually declined (P less than .05) until the end of the fast. Plasma ammonia N increased (P less than .05) and plasma albumin decreased (P less than .05) during fasting in both treatment groups, but no differences were observed between the two groups in either parameter. The data support the existence of a "glucose-alanine" cycle in that alanine decreased (P less than .05) in both groups whereas glucose remained constant and PUN increased. Together, these factors suggest enhanced gluconeogenesis from amino acids. Increased branch-chain amino acid oxidation may be responsible for the decrease (P less than .05) in total branched-chain amino acids observed during fasting. The higher dietary protein intake of lambs fed diet B apparently afforded some nutritional advantage, since most of the changes observed occurred approximately 24 hr later in these lambs than in those fed the lower protein diet A before fasting.

Lipid coating as a mode of protecting free methionine from ruminal degradation.

A method of protecting free methionine from partial ruminal degradation utilizing a lipid-protein matrix was developed. Eight wether lambs were fitted with abomasal cannulae and utilized in a 4 X 4 Latin square design experiment to determine the effectiveness of the protection matrix. The squares were blocked by animal and time, with each animal receiving each of the four diets. The diets were: (1) a negative control tall fescue and corn-based diet containing no added methionine; (2) a positive control diet that contained 3 g of methionine with ground corn, zein, coconut oil and methionine each added individually; (3) the control diet 1 supplemented with a methionine, ground corn, zein and coconut oil matrix that provided 3 g methionine/d; and (4) the control diet 1 containing the methionine matrix to provide 6 g methionine/d. Digestibility and balance data were obtained by collecting feces and urine over a 7-d period, followed by a day of blood sampling at 2, 4 and 6 h postfeeding. Abomasal samples were then subsequently collected over 3 d on a time schedule that represented every 2 h. Feeding protected methionine decreased (P less than .08) urinary N by .69 g/d and increased (P less than .08) N retention by 1.07 g/d. Plasma urea N was decreased (P less than .003) by 2.06 mg/100 ml and plasma free methionine increased (P less than .001) by 1.94 mumol/100 ml in lambs fed the protected methionine matrix. These data indicate that coating free methionine with the preparation described herein was partially effective in delivering methionine to the absorptive sites and subsequently to the tissues of the ruminant animal.

Performance and plasma amino acids of growing calves fed corn silage supplemented with ground soybeans, fishmeal and rumen-protected lysine.

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)

Bioavailability of magnesium in beef cattle fed magnesium oxide or magnesium hydroxide.

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)

Labile magnesium reserves in beef cows subjected to different prepasture supplementation regimens.

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.

Nitrogen metabolism and somatotropin secretion in beef heifers receiving abomasal arginine infusions.

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.

Effects of microbial supplements containing yeast and lactobacilli on roughage-fed ruminal microbial activities.

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.

Effect of water restriction and dietary potassium on nutrient metabolism in lambs.

Twenty-four Hampshire X Suffolk wether lambs (40 kg) were used in a 2 X 2 factorial arrangement of treatments to study the influence of water restriction and high levels of dietary K on nutrient utilization. Lambs were offered either a normal K diet (1.86% K) or a high K diet (3.66% K). Additionally, the lambs were offered either a normal quantity of water (previously determined voluntary intake of lambs fed normal K levels) or were restricted to a water intake 80% of normal. Each lamb received 800 g/d of a fescue hay-corn diet (77.5% fescue hay) with an additional 20 g of KCl offered to lambs on high K treatments. The experiment consisted of 14 d of adjustment and 7 d of total collection with blood and ruminal parameters measured on the last day of collection. Both acid detergent fiber digestibility and N balance were reduced (P less than .05) by high K intake. Urinary Mg excretion tended (P greater than .05) to be decreased by water restriction. Plasma Mg concentrations at 2 h post-feeding (PF) tended (P greater than .05) to be reduced by a high K intake at a normal level of water consumption but increased at high K intake when water intake was restricted. A similar pattern was observed for plasma K concentrations. Although high K intake resulted in elevated (P less than .05) ruminal K concentrations both at 2 and 6 h PF, ruminal concentrations of ammonia-N and volatile fatty acids were not affected (P greater than .05) by treatment. These data suggest that nutrient utilization in both the digestive tract and tissues is altered by level of K and modest water restriction.

Growth and metabolism of growing beef calves fed tall fescue haylage supplemented with protein and(or) energy.

Endophyte (Acremonium coenophialum)-infected Kentucky 31 tall fescue was fertilized in mid-August, stockpiled, harvested November 4 to 6 and stored in a concrete stave silo. Ninety-six growing calves (189 kg) were assigned to the following treatments (24 calves/treatment): 1) corn silage (CS) plus .4 kg/d of soybean meal (SBM; 2) fescue haylage plus .4 kg/d of SBM; 3) fescue haylage plus 1.6 kg/d of corn and 4) fescue haylage plus 1.6 kg/d of corn and .4 kg/d of SBM. Daily gains and dry matter (DM) intakes during the 91-d trial were .52, 4.58; .51, 5.22; .59, 6.06; and .63, 6.18 kg for treatments 1 through 4, respectively. Daily gains of calves fed corn silage plus SBM and fescue haylage plus SBM were not different (P greater than .05). However, a difference (P less than .05) existed between treatments 1 and 2 vs 3 and 4. Feed conversion was improved (P less than .05) in calves fed corn silage. Calves in a metabolism trial were fed either 1) 6.2 kg November-ensiled fescue haylage or 2) 6.2 kg November-ensiled fescue haylage plus 1.6 kg/d of corn. Digestibility of DM, N-free extract (NFE) and TDN did not differ (P greater than .05) between treatments. Ether extract digestibility was greater (P less than .05) for the added corn diet, while that of CP was greater (P less than .05) for the fescue haylage diet. Nitrogen retained was higher (P less than .05) for calves fed added corn. A follow-up trial with 96 growing calves (190 kg) compared September- and November-harvested fescue haylages supplemented with either 1.3 or 2.6 kg corn/d.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of endophyte-infected fescue on concentrations of prolactin in blood sera and the anterior pituitary and concentrations of dopamine and dopamine metabolites in brains of steers.

An experiment was conducted to determine if the decrease in circulating concentrations of prolactin in cattle consuming endophyte (Acremonium coenophialum) -infected tall fescue (Festuca arundinacea) was associated with changes in prolactin concentrations in the anterior pituitary and concentrations of dopamine (DA) and its metabolites 3,4-dihydroxyphenyl-acetic acid (DOPAC) and homovanillic acid (HVA) in the stalk median eminence (SME), preoptic area (POA) and hypothalamus (HP). Six crossbred steers that grazed high-endophyte (greater than 90% infected) fescue and four steers that grazed low-endophyte (less than 1% infected) fescue from April to September were slaughtered. Brains and pituitaries were removed and dissected. Extracts from neural tissue were analyzed for DA, DOPAC and HVA using high performance liquid chromatography/electrochemical detection. Pituitary extracts and sera from blood samples taken 5 d prior to slaughter were subjected to prolactin radioimmunoassay. Consumption of high-endophyte fescue was associated with decreased concentrations of prolactin in serum (P less than .01) and in the anterior pituitary (P = .08), decreased (P less than .05) concentrations of DA in the SME and decreased (P less than .01) concentrations of HVA in the POA and HP, but it did not influence levels of DOPAC. These results suggest that endophyte toxins may reduce prolactin synthesis and release and may alter activity of dopaminergic neurons.