Identification of methionine and lysine transport systems in cattle small intestine

Incubations of bovine ileal mucosa were used to identify the amino acid transport systems responsible for methionine and lysine uptake in cattle small intestine. Uptake was expressed as micromole per milliliter of cell water. Lysine uptake was shown to be sodium (Na) independent. Specific transport components of lysine uptake were estimated to be: 63% occurring through the cationic (Y+) system, 17% via the neutral alanine (A) and alanine, serine, cysteine (ASC) amino acid transport systems, and 20% through nonsaturable processes. Physiological concentrations of competitive inhibitors altered (P less than .05) lysine uptake. Methionine uptake was shown to occur by both Na-independent and Na-dependent transport systems. The specific transport components of methionine uptake were estimated as 54% occurring through the neutral branched chain leucine (L) systems, 28% through the ASC system, 15% through the A system and 25% through nonsaturable processes. Physiological concentrations of competitive inhibitors did not inhibit (P greater than .05) methionine uptake.     

Body and intestinal growth of broiler chicks on a commercial starter diet. 3. Development and characteristics of tryptophan transport

1. A study was conducted to characterise the development of amino acid transport in broiler chicks, using L-tryptophan as a model. The chicks were maintained on a broiler starter diet between hatch and 21 d of age. 2. There was a significant reduction in the rate of uptake of 0.04 mM L-tryptophan with age in both the jejunum and ileum. Uptake was enhanced in the presence of 50 mM sodium chloride to different degrees depending on age and intestinal site. At both intestinal sites, uptake capacity increased with age while there was a reduction in uptake efficiency with age. 3. At a concentration of 25 mM, both sodium chloride and potasium chloride increased uptake by ileal brush-border membrane vesicles (BBMV) of 7-d-old chicks but uptake was reduced when potassium chloride was included at a concentration of 50 mM. In the presence of valinomycin, uptake by jejunal BBMV was stimulated by 25 mM sodium chloride. In the presence of both sodium chloride and potassium chloride and in the absence of valinomycin, uptake was increased by 42.6% but this was reduced to 23.4% when the ionophore was included in the buffer. 4. The Na+-independent uptake of L-tryptophan into jejunal vesicles of 21-d-old chicks was lower in the presence of D-tryptophan than in the presence of 2-aminobicyclo-[2,2,1]-heptane-2-carboxylic acid (BCH). The inclusion of BCH in the incubation medium at low concentrations significantly enhanced the uptake of 0.04 mM L-tryptophan by jejunal BBMV. 5. At similar concentration (0.04 mM) to L-tryptophan, lysine, methionine and alanine in the presence of Na+ also stimulated L-tryptophan uptake. The uptake of L-tryptophan was reduced at a higher concentration, 25 mM, of these amino acids. 6. The study revealed a decline in rate of amino acid uptake and an increase in total uptake capacity with age. Tryptophan uptake was both Na+-independent and dependent, and occurred more in the ileum than in the jejunum. The uptake of L-tryptophan depended on the concentration of other amino acids and other factors in the diet and digesta.     

Localization and activities of lysosomal enzymes in jejunal and ileal epithelial cells of the young pig

The localization and the specific activities of lysosomal enzymes in jejunal and ileal epithelium of suckling pigs were determined. Acid phosphatase was found in pinocytotic vesicles and vacoles at both intestinal sites in pigs examined during (less than or equal to 1 day old) or shortly after (2 days old) closure. These pinocytolically active vesicles and vacuoles were determined to be phagolysosomes. In the ileum, phagolysosomes persisted well after closure, at an age (10 days) when ileal epithelium retains the capacity for pinocytotic uptake of immunoglobulin, but does not transport it to blood. Neither jejunum nor ileum contained phagolysosomes at an age (30 days) when the intestine had lost both uptake and transport capacities. The specific activities of acid phosphatase and cathepsin B-1 in jejunum were similar for all age groups. However, the activities of these enzymes in ileum of 10-day-old pigs were three and five times greater than at any other age or site. The results are consistent with phagolysosomes in the ileum of 10-day-old pigs functioning as a "barrier" which prevents transport of the macromolecules which are taken up pinocytotically by this epithelium. The results do not permit conclusions whether (or not) the phagolysosomes which appear in jejunum and ileum of pigs less than or equal to 1 day old contribute to closure, because the transcellular route for immunoglobulin absorption in pigs has not been precisely defined.     

Effects of chloride conductance inhibitors on fluid secretion into ligated ileal and jejunal loops in pigs

Compounds that prevent chloride transport in membrane vesicles have been tested for in vivo activity against the effects of intestinal secretory agents. Chloride channel blockers including diphenylamine-2-carboxylate, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonate, 5-nitro-2-(2-phenylethylamino)benzoic acid, and alpha-phenylcinnamic acid were tested for effects on jejunal or ileal secretion in weanling pigs. Secretion was studied in ligated intestinal loops in a control state, during exposure to secretory concentrations of theophylline, and after prior treatment with cholera toxin. Increases in net fluid flux induced by either theophylline or cholera toxin were not prevented by adding chloride channel blockers into the intestinal lumen. Channel blocker concentrations that reduced chloride transport by greater than 50% in pig jejunal brush border vesicles did not cause significant changes in unidirectional blood to lumen chloride flux measured in situ. Several routes of administration of the specific chloride channel blocker alpha-phenylcinnamate failed to reduce fluid secretion induced by theophylline. Chloride channel blocker effectiveness appears to be significantly different between in vitro and in vivo experimental models. In contrast to the chloride channel blockers, loperamide significantly reduced net fluid and chloride flux in ileal loops secreting fluid in response to theophylline. Antagonism of the production or actions of second messenger by loperamide was more effective than the chloride channel blockers in reducing conductive chloride transport associated with intestinal secretion.     

Characterization of L-lysine transport across equine and porcine jejunal and colonic brush border membrane

In nonruminant herbivores, microbially derived AA could contribute to whole-body AA homeostasis and thus decrease predicted AA requirements. However, postileal capacity of AA uptake is currently unknown. Therefore, to test the hypothesis that Lys is transported across the large colon mucosal apical membrane with capacity similar to that of the small intestinal mucosa in the pony and pig, we examined Lys transport in vitro using brush border membrane vesicles (BBMV). Mucosa was collected from the distal jejunum (DJ) and proximal large colon (PLC) of growing pigs (n = 3) and ponies (n = 4), flash frozen in liquid nitrogen, and stored at -80°C. Jejunal and colonic BBMV were manufactured by Mg(2+) precipitation and used to determine initial rates and kinetics [the maximal transport rate (V(max)) and the Michaelis constant (K(M))] of l-Lys transport into apical epithelia by rapid filtration technique in Na(+)-gradient incubation buffer. Initial rates of total l-Lys uptake did not differ between the PLC and DJ in either the pig or the pony, or between the pony and the pig, at each l-Lys concentration. In the pig, compared with the DJ, l-Lys transport V(max) in the PLC did not differ (121 ± 26 and 180 ± 26 pmol?mg of protein(-1)?s(-1), respectively; P = 0.14) and l-Lys K(M) in the PLC tended to be greater (0.23 ± 0.22 and 0.89 ± 0.22 mM, respectively; P = 0.09). In the pony, compared with the DJ, l-Lys transport V(max) in the PLC was greater (62 ± 25 and 149 ± 25 pmol?mg of protein(-1)?s(-1), respectively; P = 0.04) and l-Lys K(M) in the PLC was greater (0.08 ± 0.22 and 1.05 ± 0.22 mM, respectively; P = 0.02). l-Lysine diffusion was not different between segments; however, total intestinal diffusion was greater (P = 0.03) in the pony than in the pig (115 ± 10 and 73 ± 10 pmol·mg of protein(-1)?s(-1), respectively). These results demonstrate that the large colon is capable of l-Lys transport across the apical epithelial membrane with greater capacity and less affinity than the jejunum, indicating that the large colon may play a significant role in l-Lys absorption and homeostasis in hindgut fermenters.     

Characterization of transport systems for cysteine, lysine, alanine, and leucine in wool follicles of sheep

Aspects of the uptake of the AA Cys, Leu, Ala, and Lys into wool follicles were investigated using short-term culture of thin strips of sheep skin. Following verification of the reliability of the model system, the sites of uptake of the radiolabeled AA were shown to differ and to be consistent with their different roles in fiber production. Cysteine appeared in the zone of keratinization immediately distal to the follicle bulb. Lysine was incorporated into the germinative cells of the follicle bulb and the cells of the inner root sheath. Leucine and Ala were incorporated into the follicle bulb, inner root sheath, and keratinizing fiber. The incorporation of all AA into the dermal papilla was low. The relative rates of uptake of the AA into the wool follicle were as follows: L-Cys (100), L-Leu (5.5), L-Ala (2.5), and L-Lys (0.8). Uptake of Cys was saturable and followed Michaelis-Menten kinetics, suggesting a carrier-mediated system, with little or no diffusion. The majority (70%) of Cys uptake into follicles was via a Na-independent system that was not inhibited by alpha-(methyl-amino)isobutyric acid or 2-amino-2-norbonanecarboxylic acid and therefore is not via the normal Cys transport systems A, ASC, or L. Uptake of Cys appeared to be via a low-affinity, high-capacity transport system, which may be unique to the fiber-producing follicle. The majority of Ala transport had characteristics consistent with the functioning of system A (Na-dependent, inhibited by alpha-(methylamino)isobutyric acid, and low substrate affinity). Leucine uptake was inhibited by 2-amino-2-norbonanecarboxylic acid but was Na-dependent, suggesting that a variant of system L operates in the follicle to transport Leu. Lysine uptake was consistent with the operation of the usual Lys transporter system y+. Diets designed to maximize wool growth should provide AA profiles reflecting the relative rates of uptake demonstrated in this study. Investigations of possible polymorphisms in genes encoding AA transport proteins in follicles may reveal a source of genetic differences in wool growth potential among genotypes.     

Amino acid-supplemented raw soybean diets for finishing swine

Growth and carcass traits of finishing swine fed amino acid-supplemented raw soybean (RSB) diets were examined. Experiments 1 and 2 were identical and included the following treatments: 1) corn-soybean meal (SBM), 2) corn-RSB (lysine equal to Diet 1), 3) Diet 2 + .15% L-lysine (LYS), 4) Diet 3 + .1% L-threonine (THR), 5) Diet 3 + .05% L-tryptophan (TRP), 6) Diet 3 + .15% DL-methionine (MET) and 7) Diet 3 + THR + TRP + MET equivalent to Diets 4 to 6. The unsupplemented corn-RSB diet reduced (P less than .05) gain, gain:feed, plasma urea N, loin eye area, and percentage of muscling compared with the corn-SBM diet. The addition of LYS or LYS + TRP generally improved these response variables, whereas the addition of LYS + THR or LYS + MET tended to reduce gain and gain:feed compared with the LYS addition alone. The LYS + THR + TRP + MET addition resulted in growth and carcass traits that were intermediate between those of pigs fed the corn-RSB and the corn-SBM diets but not different (P greater than .05) from either. Experiment 3 included Diets 1, 2 and 7 from Exp. 1 and 2, in addition to the following treatment: corn-RSB + LYS + THR + TRP. The LYS + THR + TRP addition improved (P less than .05) growth and carcass traits compared with the LYS + THR + TRP + MET addition.(ABSTRACT TRUNCATED AT 250 WORDS)     

水貂不同赖氨酸、蛋氨酸水平颗粒饲料对营养物质消化代谢的影响

为确定水貂适宜氨基酸组成、降低饲料成本,试验选取成年健康公母水貂各60只,随机分成6组,采用2×3因子试验设计,即赖氨酸2个水平,分别为1.85%和1.7%,蛋氨酸3个水平分别为0.81%、0.99%和1.24%,对采食正常试验组水貂进行了消化代谢试验。结果表明:①日粮赖氨酸水平的变化对公母貂干物质摄入、排出及消化率影响不显著(P>0.05);蛋氨酸水平的变化对母貂干物质摄入、排出及消化率影响不显著(P>0.05),公貂粪干物质排出量蛋氨酸水平为0.99%组显著低于蛋氨酸水平为1.24%组(P<0.05),公貂干物质消化率蛋氨酸水平为0.99%组极显著地高于蛋氨酸水平为0.81%组和1.24%组(P<0.01)。②日粮赖氨酸水平的变化对公母貂有机物摄入、排出及消化率影响不显著(P>0.05);蛋氨酸水平的变化对公母貂有机物摄入、排出及母貂消化率影响不显著(P>0.05),公貂蛋氨酸水平为0.99%组有机物消化率显著高于蛋氨酸水平为0.81%组(P<0.05),与蛋氨酸水平为1.24%组差异不显著(P>0.05)。③公母貂蛋白质消化代谢受赖氨酸水平变化的影响不显著(P>0.05);公母貂蛋白质摄入及粪蛋白的排出受蛋氨酸水平变化影响不显著(P>0.05),蛋氨酸为1.24%时公貂尿蛋白排出显著高于0.99%水平组(P<0.05),与0.81%水平组尿蛋白排出差异不显著(P>0.05);公貂蛋白存留0.99%水平组极显著高于1.24%水平组(P<0.01);公貂蛋白质消化率0.99%水平组显著高于1.24%水平组(P<0.05),与0.81%水平组差异不显著(P>0.05);母貂蛋氨酸水平变化对各消化代谢率指标影响不显著(P>0.05)。④1.85%赖氨酸水平组公貂赖氨酸摄入、尿排出及消化代谢率均显著高于1.7%赖氨酸水平组(P<0.05),母貂间差异不显著(P>0.05),赖氨酸水平对公母貂蛋氨酸摄入、粪尿排出及消化代谢率无显著影响(P>0.05);蛋氨酸水平对公母貂赖氨酸摄入、粪尿排出无显著影响(P>0.05),蛋氨酸0.99%水平公貂赖氨酸消化代谢率均极显著高于蛋氨酸1.24%水平和0.81%水平公貂(P<0.01),蛋氨酸水平对母貂间赖氨酸消化代谢率无显著影响(P>0.05);蛋氨酸水平对公母貂蛋氨酸摄入、粪尿排出及消化代谢率均有显著影响(P<0.05),随着蛋氨酸水平的降低,蛋氨酸的摄入逐渐降低,而尿排出蛋氨酸1.24%组显著高于0.99%和0.81%水平组(P<0.05);蛋氨酸0.99%和1.24%水平组公母貂蛋氨酸的消化代谢率均极显著高于0.81%水平组(P<0.01),蛋氨酸0.99%和1.24%水平组公母貂蛋氨酸的消化代谢率无显著差异(P>0.05)。     

Methionine and lysine uptake by cattle small intestine in vitro

Incubations of bovine intestinal mucosa were characterized for location of maximal uptake of amino acid, steady-state accumulation time of the amino acids, methionine and lysine. Photomicrographs validated that cattle intestinal mucosa was isolated by the technique described. Methionine and lysine uptake was expressed as micromoles of amino acid per milliliter of cell water. The most active site of methionine and lysine uptake was the mid-ileum, while the proximal jejunum was the least active site. Both methionine and lysine uptake showed high correlation relative to intestinal region. Methionine uptake was shown to be saturated with concentrations greater than 1 mM. Lysine transport appeared to be approaching saturation at concentrations between .5 mM to 1 mM, revealing that cattle intestinal mucosa has a lower transport capacity for lysine than methionine. These results indicate the usefulness of a technique through which substrate uptake (amino acid, carbohydrate, etc.) can be characterized in cattle small intestine.     

内流质子梯度对Gly-Pro在仔猪小肠跨膜转运中的驱动作用

为研究内流质子梯度对仔猪小肠中Gly-Pro跨膜转运的影响,采用放射性同位素示踪及体外孵育技术,观察在不同H＋浓度条件下,Gly-Pro在仔猪小肠刷状缘膜囊（Brush border membrane vesicles,BBMV）的跨膜转运量。结果表明：膜囊内液pH为7.5时,Gly-Pro的转运显著地受到膜囊外液pH值的影响,pH为4.5～5.5时,转运最快;其中pH为5.0时的Gly-Pro跨膜转运速度在1-20 min一直高于pH为7.5时的转运速度;内流质子梯度的存在可以显著促进Gly-Pro的转运,无内流质子梯度时,Gly-Pro的转运不受膜囊外液pH变化的影响。研究结果提示,跨膜内流质子梯度是仔猪小肠中Gly-Pro的跨膜转运的一种驱动力。     

Response of nitrogen metabolism in preparturient dairy cows to methionine supplementation

Three multiparous Holstein cows (607 kg of BW) were surgically prepared with an elevated carotid artery and indwelling catheters in the hepatic, portal, and two mesenteric veins to study the effects of methionine supplementation on amino acid metabolism during the last 2 wk of pregnancy. The study began 15 d before the expected calving date. Dietary treatments were Control (1.53 Mcal NE(l)/kg, 15.6% CP, and 40% ruminally undegradable protein) and Control supplemented with 60 g/d of ruminally protected methionine (MET, supplying 39 g/d of DL-methionine and approximately 18 g/d of methionine available for intestinal absorption). Each cow received both dietary treatments in a crossover design. Cows were fed once daily. After 5 d on treatment, a blood flow marker (para-aminohippurate) was infused into a mesenteric vein, and arterial, portal, and hepatic blood samples were obtained at 0, 2, 6, 12, and 18 h after feeding. Net flux of methionine was calculated as the plasma arteriovenous difference multiplied by plasma flow. Dry matter intake (10.8 kg/d) and portal (824 L/h) and hepatic (995 L/h) plasma flows were not affected (P > .10) by treatment. Arterial plasma concentration of methionine was greater (P = .10) with MET (27.67 microM) than with Control (16.42 microM). Net portal absorption of methionine increased (P = .10) with MET (26.2 g/d) compared with Control (9.5 g/d). The net portal methionine flux was negatively correlated (r = -.59; P < .001) with arterial urea concentrations. Net flux of methionine across splanchnic tissues shifted (P = .06) from a net uptake with Control (4 g/d) to a net output with MET (11 g/d). Therefore, MET increased by 15 g/d the methionine supply to the rest of the body. The net uptake of methionine by splanchnic tissues observed with Control indicated a net mobilization of methionine by peripheral tissues. Results indicate that methionine was the limiting amino acid with Control and that MET was beneficial because it increased methionine supply to peripheral tissues and reduced arterial urea concentrations.     

Effects of nicarbazin on sugar intestinal absorption in rabbits.

Nicarbazin is an anticoccidial drug, used mainly in birds, which can also be used in rabbits. It has been shown to produce several effects, such as inhibition of growth and feed efficiency in poultry. The aim of the present work was to determine whether nicarbazin alters intestinal absorption of sugar. Results obtained show that nicarbazin decreases D-galactose accumulation in the jejunal tissue and increases mucosal to serosal transepithelial fluxes of this sugar, in both cases in a dose-dependent way. Furthermore, nicarbazin seems not to modify the sugar diffusion across the intestinal epithelium. The drug also stimulates the sugar uptake in brush border and basolateral membrane vesicles. The results suggest that in rabbits nicarbazin increases sugar intestinal absorption mediated by carriers.     

L-canavanine inhibits L-arginine uptake by broiler chicken intestinal brush border membrane vesicles

1. Intestinal brush border membrane vesicles (BBMV) were prepared from 3-week-old broiler chickens. 2. Electron microscopy of the BBMV fraction showed single membrane vesicles of different sizes with no electron dense material inside. No other organelles were observed. The sucrase and maltase activities were enriched by factors of 16 and 18, respectively, in the BBMV fraction in comparison with the homogenate. On the other hand, the Na+/K+-ATPase sensitivity to ouabain was increased by a factor of 0.8. 3. The BBMV showed a maximum L-[14C]-arginine uptake (944.9 +/- 22.9 pmoles/mg protein) at 45 s and thereafter it declined slowly. In the presence of 0.5 mM L-canavanine, the L-[14C]-arginine uptake by BBMV was reduced by 43.6% at 45 s. 4. It is concluded that L-canavanine inhibits L-arginine Na+-dependent transport across the enterocyte apical membrane in a highly purified intestinal BBMV from broiler chickens.     

Yeast culture supplement during nursing and transport affects immunity and intestinal microbial ecology of weanling pigs

The objectives of this study were to determine the influence of a Saccharomyces cerevisiae fermentation product on innate immunity and intestinal microbial ecology after weaning and transport stress. In a randomized complete block design, before weaning and in a split-plot analysis of a 2 × 2 factorial arrangement of yeast culture (YY) and transport (TT) after weaning, 3-d-old pigs (n = 108) were randomly assigned within litter (block) to either a control (NY, milk only) or yeast culture diet (YY; delivered in milk to provide 0.1 g of yeast culture product/kg of BW) from d 4 to 21. At weaning (d 21), randomly, one-half of the NY and YY pigs were assigned to a 6-h transport (NY-TT and YY-TT) before being moved to nursery housing, and the other one-half were moved directly to nursery housing (NY-NT and YY-NT, where NT is no transport). The yeast treatment was a 0.2% S. cerevisiae fermentation product and the control treatment was a 0.2% grain blank in feed for 2 wk. On d 1 before transport and on d 1, 4, 7, and 14 after transport, blood was collected for leukocyte assays, and mesenteric lymph node, jejunal, and ileal tissue, and jejunal, ileal, and cecal contents were collected for Toll-like receptor expression (TLR); enumeration of Escherichia coli, total coliforms, and lactobacilli; detection of Salmonella; and microbial analysis. After weaning, a yeast × transport interaction for ADG was seen (P = 0.05). Transport affected (P = 0.09) ADFI after weaning. Yeast treatment decreased hematocrit (P = 0.04). A yeast × transport interaction was found for counts of white blood cells (P = 0.01) and neutrophils (P = 0.02) and for the neutrophil-to-lymphocyte ratio (P = 0.02). Monocyte counts revealed a transport (P = 0.01) effect. Interactions of yeast × transport (P = 0.001) and yeast × transport × day (P = 0.09) for TLR2 and yeast × transport (P = 0.08) for TLR4 expression in the mesenteric lymph node were detected. Day affected lactobacilli, total coliform, and E. coli counts. More pigs were positive for Salmonella on d 7 and 14 than on d 4, and more YY-TT pigs were positive (P = 0.07) on d 4. The number of bands for microbial amplicons in the ileum was greater for pigs in the control treatment than in the yeast treatment on d 0, and this number tended to decrease (P = 0.066) between d 1 and 14 for all pigs. Similarity coefficients for jejunal contents were greater (P = 0.03) for pigs fed NY than for those fed YY, but pigs fed YY had greater similarity coefficients for ileal (P = 0.001) and cecal (P = 0.058) contents. The number of yeast × transport × day interactions demonstrates the complexity of the stress and dietary relationship.     

Na(+)-dependent transport of D-xylose by bovine intestinal brush border membrane vesicles (BBMV) is inhibited by various pentoses and hexoses

To detect whether pentoses and hexoses occurring in rumen bacteria or in hemicellulose ingested with feed and partly released in the small intestine have an affinity for the Na(+)-dependent glucose transporter of the bovine intestinal brush border membrane (BBM), we investigated whether these monosaccharides inhibit Na(+)-dependent transport of 14C-labelled D-xylose across the BBM using brush border membrane vesicles (BBMV) isolated from the mid-jejunum of cows. We used D-xylose as the transport substrate, because it has a low affinity for the Na(+)-dependent glucose transporter and thus its uptake into BBMV is more efficiently competitively inhibited by other sugars than that of D-glucose. D-Ribose, D-mannose and L-rhamnose occurring in rumen bacteria significantly inhibited Na(+)-dependent uptake of D-xylose into BBMV, but their inhibitory effect was less than that of D-glucose, D-xylose and phlorizin. This also applied to L-arabinose (and D-arabinose), which is, like D-xylose and D-galactose, a constituent of hemicellulose, and to 2-deoxy-D-glucose. Of all monosaccharides tested, only D-fructose did not affect Na(+)-dependent D-xylose transport. It is concluded that some pentoses and hexoses occurring in rumen bacteria (D-ribose, D-mannose and L-rhamnose) or hemicellulose (L-arabinose and D-xylose) have a low affinity for the Na(+)-dependent glucose transporter of the bovine BBM and may therefore be absorbed from the jejunum when released in the small intestine.     

Digestion of soybean meal and canola meal protein and amino acids in the digestive tract of young ruminants

Eight male Holstein calves (body weight 68 +/- 5 kg; age 75 +/- 6 d), each with a permanent re-entrant pancreatic cannula and T-type ileal and duodenal cannulas, were used in a crossover design with four animals per group to determine amino acid kinetics and digestibilities in the digestive tract of calves fed soybean meal (SBM) and canola meal (CM) protein. The SBM and CM diets were fed twice daily at a level of 900 g at each feeding time (air-dry basis). With the exception of methionine, crude protein and amino acid flows at the proximal duodenum, expressed as a percentage of intake, were not influenced by dietary protein source. Apparent ileal and total tract digestibilities of CP and amino acids were reduced (P less than .05) by feeding CM compared to SBM, but apparent ileal digestibility of methionine was not affected by dietary protein source. Except for methionine, net disappearance of all amino acids in the small intestine, relative to the amount fed, was higher for the SBM diet than for the CM diet. Net disappearance or synthesis of amino acids in the large intestine were not affected by dietary protein source. Similarly, dietary protein source did not affect (P greater than .05) the secretion of pancreatic juice or concentrations of protein, chymotrypsin and trypsin in pancreatic juice. Soybean meal protein has higher ileal and total gastrointestinal tract digestibility than CM protein for young, growing calves.     

Effect of season and location on semen quality and serum concentrations of luteinizing hormone and testosterone in Brahman and Hereford bulls

Hereford bulls from Montana (MH; n = 15) and Nebraska (NH; n = 15) and Brahman bulls from Texas (BB; n = 18) were relocated to one of three locations (LOC): Montana (MT), Nebraska (NE) or Texas (TX). All bulls were pubertal at the time of relocation in late May 1984. Semen was collected by electroejaculation within 1 wk after relocation and at 90-d intervals beginning in November 1984 through early February 1986. Bulls were given a GnRH challenge (200 micrograms i.m.) during the same week of semen collections. Bulls also were bled for 8 h at 20-min intervals in the fall of 1984 and the spring and fall of 1985 to determine endogenous concentrations of LH and testosterone. Season affected sperm concentration in all breeds (P less than .05) with decreases during the winter in BB and during the summer in NH and MH bulls. Brahman bulls had lower percentage of live cells (LIVE) than NH and MH bulls did (P less than .0001). Brahman bulls decreased in LIVE during the winter (P less than .001). Area under the LH curve after GnRH was lower (P less than .005) in BB than in MH and NH. Brahman bulls in MT had greater (P less than .02) area under the LH curve and lower (P less than .06) area under the testosterone curve than did BB in TX or NE during the winter. There was no seasonal fluctuation in LH or testosterone response to GnRH in NH or MH bulls at any LOC. Area under the endogenous LH curve was lowest (P less than .04) in BB. Basal endogenous testosterone concentration was greater (P less than .03) in NH than in MH or BB. Area under the endogenous testosterone curve was lower (P less than .03) in MH than in NH or BB. These results indicate that BB exhibit seasonal fluctuations in semen quality. This was not so apparent in semen quality traits of Hereford bulls. There also was a seasonal influence in BB on both endogenous testosterone and GnRH-stimulated LH and testosterone concentrations. Compared with Hereford bulls, Brahman bulls had lower endogenous and GnRH-stimulated concentrations of LH.     

Pig blastocyst development in vitro is affected by amino acids

Pig embryos were removed 5 d after onset of estrus and cultured in modified Krebs-Ringer bicarbonate (mKRB) medium containing bovine serum albumin (BSA; 1 mg/ml). Media in Exp. 1 and 2, but not in Exp. 3, contained 10% heat-inactivated lamb serum (LS). In Exp. 1, embryos were cultured in 1) mKRB + LS or mKRB + LS supplemented with 2) glutamine (2 mM); 3) phenylalanine (.1 mM), methionine (.05 mM) and isoleucine (.2 mM); or 4) phenylalanine, methionine, isoleucine and glutamine. Embryos cultured in media with supplemental phenylalanine, methionine and isoleucine attained larger (P less than .05) volumes during culture and initiated hatching at an increased (P less than .05) frequency compared with embryos cultured without additional amino acids. In contrast, adding glutamine depressed (P less than .05) the maximum volumes observed during culture. In Exp. 2 mKRB + LS was compared with mKRB + LS plus phenylalanine (.1 mM), methionine (.05 mM) and isoleucine (.2 mM); methionine and isoleucine; phenylalanine and isoleucine, or phenylalanine and methionine. Embryos cultured in media supplemented with phenylalanine and methionine attained larger (P less than .05) volumes than embryos cultured in either media without added methionine. Fewer (P less than .05) embryos cultured without methionine initiated hatching (56%) compared with embryos provided methionine (89%). In Exp. 3, we evaluated the addition of glutamine (1 mM) with or without phenylalanine (.1 mM), methionine (.05 mM) and isoleucine (.2 mM) to serum-free mKRB. Adding glutamine alone, but not in combination, increased (P less than .05) blastocyst volumes on d 3 and maximum volume attained during culture compared with embryos cultured in mKRB alone.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of dietary amino acid supplementation on measures of whole‐body and muscle protein metabolism in aged horses

The objective of this study was to examine markers of whole‐body and muscle protein metabolism in aged horses fed a diet typical for North American aged horses, supplemented with amino acids. In a replicated Latin square design, six aged horses (20 ± 1.1 years) were studied while receiving each of three isocaloric, isonitrogenous diets, a control treatment concentrate (CON; 100 mg/kg^?1 BW day^?1 lysine, 84 mg kg^?1 day^?1 threonine, 51 mg kg^?1 day^?1 methionine), LYS/THR (134 mg kg^?1 BW day^?1 lysine, 110 mg kg^?1 BW day^?1 threonine, 52 mg kg^?1 BW day^?1 methionine) and LYS/THR/MET (132 mg kg^?1 BW day^?1 lysine, 112 mg kg^?1 BW day^?1 threonine, 62 mg kg^?1 BW day^?1 methionine). In each 15‐days period, urine and faeces were collected for assessment of nitrogen balance. Blood samples were collected before and after feeding for analysis of plasma urea nitrogen (PUN), glucose, insulin and plasma amino acid concentrations. Skeletal muscle samples were collected for measurement of proteins associated with muscle protein synthesis and degradation, and horses underwent stable isotope infusion procedures for comparison of differences in whole‐body rates of protein synthesis and degradation. There was no effect of treatment on relative abundance of proteins involved in protein synthesis, nitrogen retention or phenylalanine kinetics. PUN concentrations tended to be higher for LYS/THR (p = 0.054) and were higher for LYS/THR/MET (p = 0.0056) than for CON. Atrogin‐1 abundance tended to be higher in the post‐absorptive state for the CON treatment (p = 0.07), indicating that amino acid supplementation resulted in less muscle protein degradation when horses were in the post‐absorptive state. However, lack of differences in nitrogen retention and phenylalanine kinetics indicated that whole‐body protein metabolism was not improved, and higher PUN concentrations in the supplemented diets suggest that the supplemented amino acids may have been catabolized. Amino acid availability was not limiting protein synthesis in the sedentary aged horses in this study when fed the CON diet.