Biomedical subjects

G E Lobley

Publications and source records attributed to G E Lobley.

At least 19 recordsLinked to original sources

PubMed2007

Effect of protein supply on hepatic synthesis of plasma and constitutive proteins in lactating dairy cows.

The effects of metabolizable protein (MP) supply on the synthesis of plasma total proteins and albumin, as well as total hepatic protein synthesis, were determined in 6 multicatheterized lactating Holstein cows. Three TMR formulated to supply the same amount of energy but different amounts of MP, 1,922 (low), 2,264 (medium), and 2,517 g of MP/d (high), were fed every 2 h according to a double 3 x 3 Latin square design. For the low and high MP treatments, the cows were continuously infused with [(2)H(5)]Phe (d5-Phe) into a jugular vein for 8 h (1.3 mmol/h) on d 21 of each period. Concentration and isotopic enrichment of d5-Phe were measured for free plasma Phe, plasma total proteins, and albumin on hourly samples collected between 3 and 8 h. Low MP decreased the plasma albumin concentration (32.3 vs. 33.7 +/- 0.11 g/L) but the plasma total protein concentration was unchanged (74.1 vs. 75.6 +/- 1.13 g/L). Incorporation of d5-Phe over time into both plasma total proteins and albumin was linear (R(2) > 0.98). Neither fractional nor absolute synthesis rates of plasma total proteins (6.8 vs. 6.5 +/- 0.65%/d; 168 vs. 154 +/- 19.9 g/d) or albumin (3.4 vs. 3.4 +/- 0.10%/d; 36.3 vs. 36.5 +/- 1.11 g/d) were affected by the MP supply. Net hepatic removal of Phe was lower with the low-MP diet (-12.3 vs. -20.2 +/- 1.98 mmol/h). As a result, net hepatic Phe removal used for total export protein synthesis (17.9 vs. 11.1 +/- 1.83%) and albumin synthesis (4.6 vs. 2.9 +/- 0.54%) tended to be greater at low MP. These results suggest that hepatic synthesis of plasma proteins, including albumin, is maintained in lactating dairy cows even when the protein supply is reduced.

Animal Nutritional Physiological Phenomena↗

PubMed2006

Hepatic metabolism of 2-hydroxy-4-methylthiobutyrate in growing lambs.

This study was undertaken to determine how, and where, 2-hydroxy-4-methylthiobutyrate (HMTBA) can augment Met metabolism in lambs. Four lambs (initial body weight of 50 kg, SE = 2, and 6 mo of age) prepared with catheters in the mesenteric, portal, hepatic, and jugular veins plus the aorta, were fed at 1.5x maintenance on a grass hay, barley, fish meal, molasses/pre-mix (5:3:1:1, as fed) diet, supplied as hourly meals. Lambs were infused for 10 h with [methyl-2H3]Met (0.11 mmol/h) in a jugular vein and p-aminohippurate into the mesenteric vein. From 1 h onwards, successive 3-h infusions of saline (control), 0.55 mg/min (3.67 micromol/min), and 4.44 mg/min (29.6 micromol/min) of HMTBA were also infused into the mesenteric vein. Plasma, sampled continuously, was collected every 20 min during the last 60 min of each infusion. All infused HMTBA was recovered at the portal vein with 25% extracted subsequently by the liver. Portal appearance of total Cys and Met was unaltered by HMTBA infusion, but net splanchnic appearance of Cys increased (0.04, 0.08, 0.23 mmol/h, SEM = 0.05), whereas Met decreased (0.14, -0.01, -0.21 mmol/h, SED = 0.05). Despite this, arterial Met increased (27.0, 30.7, 51.5 microM, SEM = 2.1) as did Met irreversible loss rate (27.6, 28.7, 40.1 micromol/h, SEM = 0.51), equivalent to 40% of the HMTBA reentering the plasma after conversion to Met. These data indicate that, in ruminants, HMTBA is probably converted to Met within peripheral tissues; that is, where the metabolic need for Met exists.

Animals↗

PubMed2006

Absorption of 2-hydroxy-4-methylthiobutyrate and conversion to methionine in lambs.

Absorption and metabolism of the Met hydroxy analog 2-hydroxy-4-methylthiobutyrate (HMTBA) was examined using stable isotopes. In the first trial, Dl[1-13C]HMTBA was infused for 6 h (7.4 micromol/min) into the abomasum, and [2H3]Met was infused into the mesenteric vein, of 4 lambs prepared with vascular catheters across the splanchnic bed. Daily, lambs were offered 35 g of a mixed forage-concentrate feed/kg. Recovery of HMTBA at the portal vein was 87%, and of this, 63% bypassed the liver. In contrast, hepatic extraction of Met equaled or exceeded net absorption. Only small quantities of Met synthesized from HMTBA were exported from either the digestive tract or liver, but there was substantial and significant input from posthepatic tissues. In a second experiment, 3 of the lambs were killed following 4-h infusions of DL[1-13C]HMTBA and [2H3]Met with enrichments monitored in 15 tissues. Only kidney showed [1-13C]Met enrichment higher than plasma, which suggests that it must be a primary source of plasma Met derived from HMTBA. Based on comparison of plasma and intracellular [1-13C]:[2H3]Met enrichments, all tissues synthesized Met from HMTBA but to significantly different extents. The lowest values were for muscle, skin, brain, and lung; intermediate conversions occurred in rumen, omasum, abomasum, duodenum, jejunum, ileum, and cecum; and the greatest synthesis, equivalent to 22 to 24% of Met entry into cells, was observed for liver and kidney. Therefore, although liver and kidney both converted HMTBA to Met, it was retained by the former and exported by the latter. Under these experimental conditions, synthesis of Met from HMTBA completely eliminated use of dietary Met.

Abomasum↗

PubMed2006

What is the true supply of amino acids for a dairy cow?

Improving the prediction of milk protein yield relies on knowledge of both protein supply and requirement. Definition of protein/amino acid supply in ruminants is a challenging task, due to feedstuff variety and variability and to the remodeling of nutrient intake by the rumen microflora. The questions arise, therefore, how and where should we measure the real supply of AA in the dairy cow? This review will follow the downstream flow of AA from duodenum to peripheral tissue delivery, with a glance at the efficiency of transfer into milk protein. Duodenal AA flow comprises rumen undegradable feed, microbial protein, and endogenous secretions. Most attention has been directed toward definition of the first two contributions but the latter fraction can represent as much as 20% of duodenal flow. More information is needed on what factors affect its magnitude and overall impact. Once digested, AA are absorbed into the portal vein. The ratio of portal absorption to small intestinal apparent digestion varies among essential AA, from 0.43 (threonine) to 0.76 (phenylalanine), due to the contributions of preduodenal endogenous secretions to the digestive flow, non-reabsorption of endogenous secretions and gut oxidation of AA. Few data are available on these phenomena in dairy cows but the evidence indicates that they alter the profile of AA available for anabolic purposes. Recent comparisons of estimated duodenal flux and measured portal flux have prompted a revisit of the NRC (2001) approach to estimate AA flows at the duodenum. Changes to the model are proposed that yield predictions that better fit the current knowledge of AA metabolism across the gut. After absorption, AA flow first to the liver where substantial and differential net removal occurs, varying from zero for the branched-chain AA to 50% of portal absorption for phenylalanine. This process alters the pattern of net supply to the mammary gland. Overall, intermediary metabolism of AA between the duodenum and the mammary gland biologically explains the decreased efficiency of the transfer of absorbed AA into milk protein as maximal yield is approached. Therefore, variable, rather than fixed, factors for transfer efficiencies must be incorporated into future predictive models.

Absorption↗

PubMed2006

Absorption and digestive tract metabolism of 2-hydroxy-4-methylthiobutanoic acid in lambs.

Anabolic availability of the hydroxyl methionine analog, 2-hydroxy-4-methylthiobutanoic acid (HMTBA), given as oral doses to lambs, was quantified both directly as appearance in the portal vein and as synthesis to Met by digestive tract tissues. Eight lambs, prepared with vascular catheters in the mesenteric and portal veins plus the aorta, received twice daily for 7 d either 0.46 g or 2 g of HMTBA. On d 7, [1-13C]HMTBA was supplied as 1 oral dose while [methyl-2H3]Met was infused into the jugular vein. Peak absorption as HMTBA occurred 70 to 90 min after the oral dose. All digestive tract tissues converted HMTBA to Met, equivalent to 24% of the Met provided by the diet for the larger HMTBA dose. Overall, total availability of HMBTA averaged 17.9% of the dose (range 10.6 to 27.9%), with 12.5% (range 7 to 22%) as absorbed HMBTA and the remainder as Met synthesized by digestive tract tissues. Release of 13CO2 into the portal vein accounted for another 23% of the dose. In all digestive tract tissues, the d-isomer was present in a smaller proportion than in the dose. In terms of whole-body kinetics, HMTBA loss from the plasma followed first-order kinetics, with a mean biological half-life of 76 min. Using this value, a simple model was devised to estimate HMTBA absorption based on peripheral plasma samples. When compared with direct measures of absorption, the model gave a slope of 0.81 (R2 = 0.68) and offers a practical means to test HMTBA availability to animals.

Absorption↗

PubMed2006

Effect of casein and propionate supply on mammary protein metabolism in lactating dairy cows.

The effects of casein (CN) and propionate (C3) on mammary AA metabolism were determined in 3 multiparous Holstein cows fitted with both duodenal and ruminal cannulas and used in a replicated Youden square with six 14-d periods. Casein (743 g/d in the duodenum) and C3 (1,041 g/d in the rumen) infusions were tested in a factorial arrangement. For each period, L-[1-(13)C]Leu (d 11) and NaH[13C]O3 (d 13) were infused into a jugular vein, and blood samples were taken from the carotid artery and the mammary vein to determine Leu kinetics and net uptake of AA. Both CN and C3 treatments separately increased milk protein concentration and yield. With CN there was a general response in mammary protein metabolism, involving increases in Leu net uptake (30%), the uptake:output ratio (8%), protein synthesis (11%), secretion in milk protein (21%), and oxidation (259%). In contrast, C3 treatments tended to increase only Leu in milk protein (7%) and, when in combination with CN, to reduce Leu used for protein synthesis (5%). Across all treatments, most Leu uptake by the mammary gland was accounted for as Leu in milk or oxidized, and the Leu balance was therefore achieved without involvement of either net peptide use or production. Mammary uptake of group 1 AA increased to match milk output with all infusions. In contrast, mammary uptake of group 2 AA exceeded output to a greater extent with CN than with C3 infusions, whereas the increment in uptake of group 3 AA increased with C3 treatments. Overall, these data suggest that different mechanisms operate to improve milk protein production when either protein or energy is supplied.

Amino Acids↗

PubMed2006

Effect of postruminal glutamine supplementation on immune response and milk production in dairy cows.

Seventeen multiparous Holstein cows were used to examine the effect of an increased duodenal supply of Gln on immune function and production. Cows received continuous abomasal infusions of water (control: n = 8) or 300 g/d of Gln (n = 9) for 21 d starting within 48 h of calving. There were nonsignificant increases in milk and milk protein yields in response to Gln supplementation. Glutamine treatment had no effect on plasma glucose, nonesterified fatty acids (NEFA), or beta-hydroxybutyrate (BHBA) concentrations but did tend to increase plasma urea N concentration. The Gln treatment resulted in an increase of 108 microM in the plasma Gln concentration. Total essential AA concentrations decreased with the Gln treatment, whereas total nonessential AA concentrations were unaffected. T Lymphocyte proliferation did not differ between the control and Gln-treated cows. Treatment had no effect on the relative abundance of CD8 T cells but did increase the abundance of CD4 T cells. Cytokine production, as measured by IFN-gamma concentration determined in vitro in concanavalin-A-stimulated peripheral blood mononuclear cells, was similar between the treatments. Over the first 3 wk following calving, Gln supplementation had limited effects on milk production, metabolic parameters, and immune function.

3-Hydroxybutyric Acid↗

PubMed2005

Effects of dietary supplements of folic acid and rumen-protected methionine on lactational performance and folate metabolism of dairy cows.

The present experiment was undertaken to determine the interactions between dietary supplements of folic acid and rumen-protected methionine on lactational performance and on indicators of folate metabolism during one lactation. Fifty-four multiparous Holstein cows were assigned to 9 blocks of 6 cows each according to their previous milk production. Within each block, 3 cows were fed a diet calculated to supply methionine as 1.75% metabolizable protein, equivalent to 70% of methionine requirement, whereas the 3 other cows were fed the same diet supplemented with 18 g of a rumen-protected methionine supplement. Within each diet, the cows received 0, 3, or 6 mg/d of folic acid per kg of body weight. Rumen-protected methionine increased milk total solid concentration but not yield. Supplementary folic acid increased crude protein and casein concentrations in milk of cows fed no supplementary methionine and the effect increased as lactation progressed; it also decreased milk lactose concentration. Folic acid supplements had the opposite effects on milk crude protein, casein, and lactose concentrations in cows fed rumen-protected methionine. Milk and milk component yields and dry matter intake were unchanged. Folic acid supplementation increased serum folates and this response was greater at 8 wk of lactation. It decreased serum cysteine in cows fed rumen-protected methionine, whereas it had no effect in cows fed no supplementary methionine. The highest serum concentrations of cysteine but the lowest of vitamin B(12) were observed at 8 wk of lactation. Serum clearance of folic acid following an i.v. injection of folic acid was slower at 8 wk of lactation. During this period, the high concentrations of serum folates and cysteine, the low serum concentrations of vitamin B(12) and methionine, and the slow serum clearance of folates strongly suggest that the vitamin B(12) supply was inadequate and interfered with folate use. It could explain the limited lactational response to supplementary folic acid observed in the present experiment.

Animals↗

PubMed2004

Effect of plasma insulin and branched-chain amino acids on skeletal muscle protein synthesis in fasted lambs.

The increase in fractional rate of protein synthesis (Ks) in the skeletal muscle of growing rats during the transition from fasted to fed state has been explained by the synergistic action of a rise in plasma insulin and branched-chain amino acids (BCAA). Since growing lambs also exhibit an increase in Ks with level of feed intake, the objective of the present study was to determine if this synergistic relationship between insulin and BCAA also occurs in ruminant animals. Six 30 kg fasted (72 h) lambs (8 months of age) received each of four treatments, which were based on continuous infusion into the jugular vein for 6 h of: (1) saline (155 mmol NaCl/l); (2) a mixture of BCAA (0.778 micromol leucine, 0.640 micromol isoleucine and 0.693 micromol valine/min.kg); (3) 18.7 micromol glucose/min.kg (to induce endogenous insulin secretion); (4) co-infusion of BCAA and glucose. Within each period all animals received the same isotope of phenylalanine (Phe) as follows: (1) L-[1-13C]Phe; (2) L-phenyl-[ring 2H5]-alanine; (3) L-[15N]Phe; (4) L-[ring 2,6-3H]Phe. Blood was sampled serially during infusions to measure plasma concentrations of insulin, glucose and amino acids, and plasma free Phe isotopic activity; biopsies were taken 6 h after the beginning of infusions to determine Ks in m. longissimus dorsi and vastus muscle. Compared with control (saline-infused) lambs, Ks was increased by an average of 40% at the end of glucose infusion, but this effect was not statistically significant in either of the muscles sampled. BCAA infusion, alone or in combination with glucose, also had no significant effect on Ks compared with control sheep. Ks was approximately 60% greater for vastus muscle than for m. longissimus dorsi (P<0.01), regardless of treatment. It is concluded that there are signals other than insulin and BCAA that are responsible for the feed-induced increase in Ks in muscle of growing ruminant animals.

Amino Acids, Branched-Chain↗

PubMed2004

Effect of level of metabolizable protein on splanchnic flux of amino acids in lactating dairy cows.

The response of splanchnic tissue metabolism to different levels of metabolizable protein (MP) was measured in 6 catheterized multiparous lactating Holstein cows. Three diets, balanced to provide similar energy intakes and increasing amounts of MP (g/d)-1922 (low), 2264 (medium), and 2517 (high)-were fed during 21-d experimental periods according to a replicated Latin square. On d 18, 19, or 20, six hourly blood samples were collected simultaneously from the portal and hepatic veins plus an artery to determine net fluxes of nutrients across the portal-drained viscera and the liver. Yields of milk and protein increased, as did urinary N excretion with increasing MP. Portal absorption of essential amino acids (EAA) increased linearly with increasing MP supply, as did liver removal of His, Met, and Phe. In contrast, liver removal of the branched-chain AA (BCAA) and lysine was unaffected by diets. With increasing MP, the ratio of milk output to postliver supply of BCAA, Thr, and Lys decreased linearly, indicating oxidation of these AA in the peripheral tissues. Concomitant to a decreased catabolism of EAA in the liver (His, Met, Phe, and Thr) and/or in peripheral tissues (BCAA, Lys, and Thr), the efficiency of transfer of absorbed EAA into milk protein decreases markedly as protein supply increases. The efficiency of transfer of absorbed AA into milk also varies greatly between AA. These 2 important factors should be taken into account when building predictive schemes for milk protein output.

Amino Acids↗

PubMed2003

Whole-body protein turnover of a carnivore, Felis silvestris catus.

The cat (Felis silvestris catus) has a higher dietary protein requirement than omnivores and herbivores, thought to be due to metabolic inflexibility. An aspect of metabolic flexibility was examined with studies of whole-body protein turnover at two levels of dietary protein energy, moderate protein (MP; 20 %) and high protein (HP; 70 %), in five adult cats in a crossover design. Following a 14 d pre-feed period, a single intravenous dose of [15N]glycine was administered and cumulative excretion of the isotope in urine and faeces determined over 48 h. N flux increased (P<0.005) with dietary protein, being 56 (se 5) mmol N/kg body weight (BW) per d for cats fed the MP diet and 146 (se 8) mmol N/kg BW per d for cats fed the HP diet. Protein synthesis was higher (P<0.05) on the HP diet (75 (se 10) mmol N/kg BW per d; 6.6 (se 1) g protein/kg BW per d) than the MP diet (38 (se 5) mmol N/kg BW per d; 3.4 (se 0.4) g protein/kg BW per d). Protein breakdown was higher (P<0.05) on the HP diet (72 (se 8) mmol N/kg BW per d; 6.3 (se 0.7) g protein/kg BW per d) than the MP diet (44 (se 3) mmol N/kg BW per d; 3.9 (se 0.3) g protein/kg BW per d). Compared with other species the rate of whole-body protein synthesis in the well-nourished cat (9.7 (se 1.3) g protein/kg BW0.75 per d) is at the lower end of the range. These results show that feline protein turnover adapts to dietary protein as has been shown in other species and demonstrates metabolic flexibility. Further work is required to determine exactly why cats have such a high protein requirement.

Adaptation, Physiological↗

PubMed2003

Effect of feed intake on amino acid transfers across the ovine hindquarters.

Responses in variables of amino acid (AA) metabolism across peripheral tissues to feed intake were studied in six sheep (mean live weight 32 kg) prepared with arterio-venous catheters across the hindquarters. Four intakes (0.5, 1.0, 1.5 and 2.5 x maintenance energy) were offered over 2-week periods to each sheep in a Latin square design with two animals replicated. Animals were infused intravenously with a mixture of U-13C-labelled AA for 10 h and integrated blood samples withdrawn from the aorta and vena cava hourly between 5 and 9 h of infusion. Biopsy samples were also taken from skin and m. vastus lateralis. Data from both essential (histidine, isoleucine, leucine, lysine, phenylalanine, threonine) and nonessential (glycine, proline, serine, tyrosine) AA were modelled to give rates of inward and outward transport, protein synthesis and degradation, plus the fraction of total vascular inflow that exchanged with the hindquarter tissues. Rates of inward transport varied more than 10-fold between AA. For all essential AA (plus serine), inward transport increased with food intake (P<0.04). There were corresponding increases in AA efflux (P<0.05) from the tissues for threonine and the branched-chain AA. Protein synthesis rates estimated from the kinetics of these AA also increased with intake (P<0.02). Rates of inward transport greatly exceeded the amount of AA necessary to support protein retention, but were more similar to rates of protein synthesis. Nutritional or other strategies to enhance AA transport into peripheral tissues are unlikely to increase anabolic responses.

Amino Acids↗

PubMed2002

Glutamine metabolism in ovine splanchnic tissues: effects of infusion of ammonium bicarbonate or amino acids into the abomasum.

This study investigates the effects of increased NH3 or amino acid supply on glutamine utilisation and production by the splanchnic tissues of fed sheep. Six sheep, prepared with vascular catheters in the aorta, mesenteric, portal and hepatic veins, were fed grass pellets to 1.1 x energy maintenance requirements. Each treatment involved a 4 d abomasal infusion, of either ammonium bicarbonate (AMM; 234 micromol/kg(0.75 per min), water (CONT), or a mixture of amino acids that excluded glutamine and glutamate (AA; 46.8 micromol amino acid-N/kg(0.75) per min). The treatments simulated nutritional extremes in terms of the balance of absorbed N. Kinetics across the whole gut and the liver were monitored during an intra-jugular infusion of [5-(15N)]glutamine. Blood flow across the whole gut or liver were unaffected by treatment. Both AMM and AA infusions doubled the hepatic release of urea-N compared with CONT (P<0.02). AA infusion decreased arterial glutamine concentration by 26% (P<0.01) and 23 % (P<0.05) compared with AMM and CONT respectively. Despite this, whole-body glutamine flux was not affected by treatment. In contrast, AMM infusion increased hepatic glutamine production by 40% compared with CONT (P<0.02). This provided a mechanism to ensure NH3 supply to the periphery was maintained within the normal low physiological levels. Hepatic glutamine utilisation tended to increase during AA infusion, probably to ensure equal inflows of N to the ornithine cycle. Between 6 and 10% of NH3 absorbed across the digestive tract was derived from the amido-N of glutamine. Overall, splanchnic glutamine utilisation accounted for 45-70% of whole-body glutamine flux.

Abomasum↗

PubMed2002

Effect of supply of metabolizable protein on splanchnic fluxes of nutrients and hormones in lactating dairy cows.

The effect of the supply of metabolizable protein on splanchnic fluxes of nutrients and hormones was measured in six catheterized late-lactation Holstein cows in a crossover design. Two isonitrogenous diets (16.3% CP), but differing in rumen protein degradability and estimated metabolizable protein (MP) supply (1654 g/ d, Lo-MP; 1930 g/d, Hi-MP) were fed, each over a 35-d experimental period. On d 34 or 35, net fluxes of nutrients and hormones across the portal-drained viscera, the liver, and total splanchnic tissues were determined. Portal absorption of total, essential, nonessential, and branched-chain amino acids (AA) increased with the Hi-MP diet. Approximately 76% of the additional metabolizable protein supply was recovered as extra AA-N absorption in the portal vein. Liver removal of AA was not different between diets, and this resulted in a greater net release across the splanchnic tissues for the Hi-MP diet. This extra AA supply provided substrates for the observed increased milk protein yield for the Hi-MP diet. Fractional efficiencies of conversion of absorbed individual essential AA into milk protein ranged from 0.42 to 0.68. The corresponding efficiencies for utilization of postsplanchnic AA supply were 0.42 to 1.80. Provision of methionine, phenylalanine, and histidine beyond the liver were similar to outputs in milk protein but the other essential AA were supplied to peripheral tissues in excess of milk output, indicative of oxidative mechanisms in nonhepatic tissues. Net fluxes of glucose, NH3-N, and urea were not affected by the diets. Neither arterial concentrations of insulin, somatotropin, or IGF-1, nor net transfers across the portal-drained viscera or liver of insulin, were affected by the diets. Although portal release of glucagon was not different between the diets, a smaller proportion was removed by the liver on the Hi-MP diet. Metabolism of AA across the splanchnic tissue bed is a major determinant of the quantity and the profile of AA delivered to peripheral tissues.

Amino Acids↗

PubMed2002

Effect of supply of metabolizable protein on whole body and splanchnic leucine metabolism in lactating dairy cows.

The effect of the supply of metabolizable protein (MP) on protein metabolism across the splanchnic tissues was determined in six catheterized lactating Holstein cows. In a crossover design, two isonitrogenous (16.3% CP) diets balanced to provide a low (Lo-MP) or high (Hi-MP) supply of MP were fed over 35-d periods. After 24 d of feeding, N balance was determined over a 6-d period. On d 33, [13C] sodium bicarbonate was infused into one jugular vein for 6 h, and hourly breath samples were collected. On d 34 or 35, L[1-(13)C] leucine was infused into one jugular vein, and between 2 to 6 h of infusion, breath and blood samples were taken hourly from the portal and hepatic veins and an artery. Isotopic enrichments of plasma leucine, 4-methyl-2-oxopentanoate, and expired CO2 were determined for calculation of leucine kinetics. Net leucine absorption was greater, either on a direct basis (leucine transfer only) or corrected for portal-drained viscera metabolism to 4-methyl-2-oxopentanoate and CO2 for the Hi-MP diet. There were no effects of diet on hepatic net flux of leucine across the liver, and, thus, more leucine was available to peripheral tissues with the Hi-MP diet. Combined with an increment in portal absorption of most of essential AA, this led to increased milk protein output, although it only represented 16% of the additional available leucine. Whole body leucine oxidation was also greater for the Hi-MP diet, as was leucine used for protein synthesis. Despite these changes, MP supply did not affect irreversible loss rate of leucine by portal-drained viscera and the liver; these averaged 35 and 20% of whole body irreversible loss rate, respectively. These ratios confirm the high metabolic activity of splanchnic tissues in lactating dairy cows, which are even greater than previously reported in growing ruminants.

Animal Nutritional Physiological Phenomena↗

PubMed2002

Effect of dietary fiber on endogenous nitrogen flows in lactating dairy cows.

The effect of dietary fiber on endogenous N secretion was studied using a 15N isotope dilution technique in four fistulated Holstein cows. Two isonitrogenous diets differing only in fiber (NDF and ADF) content were used in a crossover design. One diet (HF) contained 37.4% NDF, while the other (LF) contained 23.3%. A new model was developed to estimate endogenous N secretions and losses for the preintestinal, intestinal, and the total sections of the gastrointestinal tract. Three precursor pools: TCA-soluble fraction of plasma, intestinal mucosa, and milk were compared. Although endogenous losses estimated with the model were numerically different for each precursor pool selected (TCA-soluble fraction > mucosa > milk), treatment effects were similar. As intestinal mucosa is probably closest to the precursor pool, these data are discussed. Non-urea N endogenous secretions contributed 13% of the duodenal N flow but were not affected by the fiber content of the diet. The nonurea N endogenous flow at the duodenum was comprised of approximately equal inputs from endogenous N direct, and that incorporated into the microbial biomass. Total endogenous N flows at the duodenum exceeded, by nearly twofold, estimated inputs of urea-N to microbial biomass. Metabolic fecal output averaged 17% of fecal N and was not affected by level of dietary fiber, but net losses from secretions occurring in the small intestine were higher with the low fiber diet. Overall, endogenous N secretions represented 30% of total digestive tract protein synthesis.

Animals↗

PubMed2001

Lysine metabolism across the hindquarters of sheep; effect of intake on transfers from plasma and red blood cells.

Both plasma and red blood cells contain amino acids (AA), but the relative amount of AA transferred from each vascular compartment to the tissues remains unclear. For splanchnic tissues, the relative transfers between the plasma, the red blood cells and the tissues may vary with nutritional state, but whether the same situation pertains for other tissues is not known. The current study focused on the transfer of lysine from plasma and red blood cells across the hindquarters of sheep offered four levels of intakes (0.5, 1.0, 1.5 and 2.5 x maintenance energy). This design, coupled with use of [U-13C]lysine as tracer, also allowed the effect of intake on protein kinetics to be examined. At all intakes, the concentration of lysine in the sheep' red blood cells exceeded that in plasma by 50% (P<0.001), while the distribution of labelled lysine between the plasma and the red blood cells was 0.71:0.29. Net lysine uptake by the hindquarters increased in a linear manner (P<0.001) with intake, with more than 90% extracted from the plasma. Free lysine enrichments in plasma from the posterior vena cava were less than that from the artery (P<0.001), but those in red blood cells were not different between the artery and vein. The red blood cells thus play a minor role in the transfers to and from the hindquarter tissues, regardless of intake. Based on plasma transfers and the enrichment of lysine in arterial plasma, hindquarter protein synthesis increased linearly with intake (P<0.001). In contrast, protein breakdown was unaffected by intake. The contribution of hindquarter protein synthesis to whole-body lysine flux remained unchanged with intake (18-20%).

Analysis of Variance↗

PubMed2001

Does glutamine act as a substrate for transamination reactions in the liver of fed and fasted sheep?

The present study investigated the relative importance of glutamine as a transamination source in the ovine liver by examination of the labelling of amino acids (AA) in the hepatic free pool, mixed liver and plasma proteins of fed and fasted sheep, following infusion of isotopically-labelled glutamine. In a cross-over design four sheep were either fasted for 3 d or fed to maintenance and finally euthanased. At each intake, the sheep were infused for 6 h with [2-15N]glutamine (150 micromol/h) and samples of total plasma protein isolated. Following the terminal infusion, liver tissue total proteins were prepared and hydrolysed and 15N-enrichments in seventeen AA were determined by GC-combustion-isotope-ratio mass spectrometry. All AA were enriched (relative to natural abundance) except lysine and threonine, with the lowest enrichments in phenylalanine and histidine. There was no effect of the fed v. fasted state, except for leucine and isoleucine in liver protein Enrichments in liver protein were greater than in plasma protein except proline) and probably reflect the faster turnover rate of hepatic constitutive proteins compared with export proteins. Amination to methionine was greater than that to phenylalanine suggesting a mechanism for preferentially protecting the former. This factor could be important for ruminant production, as methionine is often considered to be the first limiting AA for animals offered certain silages and conserved forages. Enrichments in all AA (except for glutamine, alanine and aspartate) were less than that for glutamate and thus transaminations may have occurred with glutamine directly or via glutamate, following the action of hepatic glutaminase.

Analysis of Variance↗