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L P Milligan

Publications and source records attributed to L P Milligan.

At least 19 recordsLinked to original sources
PubMed1995

In vitro ouabain-sensitive respiration and protein synthesis in rumen epithelial papillae of Hereford steers fed either timothy hay or timothy hay supplemented with cracked corn once daily.

Rumen epithelial papillae samples, acquired from the Central sac of six adult Hereford steers (585 +/- 17 kg) fed either timothy hay plus soybean meal (T) or timothy hay plus cracked corn and soybean meal (TC) once daily (0900), were used in a crossover design to study the daily pattern of O2 consumption and protein synthesis. Tissue samples were acquired at 0900, 1200, 1800, and 2400 over a 10-d sampling period. Additionally, ruminal fermentation characteristics (pH, ammonia, VFA, and osmolality) were measured at 0430, 0900, 1030, 1200, 1500, 1800, 2100 and 2400. Total, ouabain-insensitive (OIO2), and cycloheximide-insensitive (CIO2) O2 consumption were greatest at 2400 (P < .01, P < .06, and P < .02 respectively). Additionally, steer fed TC had a greater CIO2 at 2400. In conjunction with a temporal effect on ruminal fermentation patterns, after an initial decline, there was an increase (P < .05) in total O2, ouabain-sensitive (OSO2), OIO2, and CIO2 consumption throughout the day. Fractional rates of protein synthesis were not different at any time point between diets. Rumen epithelial metabolism exhibits a temporal pattern in cattle fed once daily.

Ammonia↗
PubMed1994

Production of 2-aminobutyrate by Megasphaera elsdenii.

Production of the amino acid 2-aminobutyrate was studied in four strains of Megasphaera elsdenii grown on a lactate-based growth medium containing Bacto-casamino acids and yeast extract. Supplementation with threonine increased the production of 2-aminobutyrate in three of the four strains, but no substantial increase in production was noted with serine, methionine, or aspartate, all of which are potential sources for the precursor of 2-aminobutyrate, 2-oxobutyrate. L-Cycloserine, an inhibitor of alanine transaminases, decreased both alanine and 2-aminobutyrate production, suggesting that 2-aminobutyrate synthesis may share the same metabolic pathway as alanine synthesis or that 2-oxobutyrate can act as a substrate for alanine transaminases. Decreases in the production of 2-aminobutyrate were associated with a reduction in the catabolism of branched-chain amino acids in two of the four strains.

Amino Acids↗
PubMed1993

In vitro ouabain-sensitive respiration and protein synthesis in ruminal epithelial papillae of Hereford steers fed either alfalfa or bromegrass hay once daily.

Seven Hereford steers (457 +/- 8.5 kg BW) with ruminal cannulas were fed either long or short chopped bromegrass (B) or alfalfa (A) hay at 90% of a 3-h ad libitum intake once daily between 0900 and 1200 in a 4 x 4 complete and 3 x 4 incomplete Latin square design. After evacuation of ruminal contents, papillae from the ventral sac of the rumen were excised at 0900, 1200, and 2100 and placed in oxygenated media. Oxygen consumption was determined polarographically, after which either 10(-4) M ouabain or 10(-4) M cycloheximide was added for determination of ouabain-sensitive (OSO2) or cycloheximide-sensitive (CSO2) O2 consumption, respectively. Additionally, protein synthesis was measured by uptake of [3H]phenylalanine. Twenty-four-hour patterns of ruminal fluid pH, osmolality, ammonia N, and VFA were also determined. Steers fed A exhibited a rapid rise in total O2, OSO2, and CSO2 immediately after consumption of a meal; the respiration patterns of ruminal epithelial papillae from animals fed B lagged behind those observed for animals fed A. Patterns of O2 consumption for both diets paralleled those observed for ruminal concentration of products of fermentation; those fed A had a larger magnitude of change in both O2 consumption and fermentation products. Ruminal epithelial O2 consumption seems to be determined by substrate availability and products of fermentation, and Na+,K+ATPase and protein synthetic activity each account for one-fifth of ruminal papillae O2 consumption. Fractional rates of protein synthesis were unaffected by type of forage consumed.

Ammonia↗
PubMed1993

In vivo nuclear magnetic resonance spectroscopy of chicken embryos from two broiler strains of varying fat content.

In vivo nuclear magnetic resonance (NMR) imaging and spectroscopy techniques were used to monitor changes in P- and H-containing molecules in embryos of two broiler strains (30 and 31) differing genetically in fat content and ranging in age from 0 to 20 days of incubation. Chemical analysis showed that Strain 30 has more carcass fat than Strain 31 at market age (7 wk). Proton (1H) and 31P spectra were obtained on four eggs per strain at Days 0, 2, 4, 6, 8, 11, 12, 14, 16, 17, 19, and 20 of incubation. Fat:water, phosphomonoester (PME):phosphodiester (PDE), and adenosine triphosphate (ATP):PDE ratios were calculated. Chicks were hatched, grown to market weight (2,000 g for females and 2,300 g for males at 7 wk), and the whole intact carcasses were analyzed for crude fat. Hydrogen-1 NMR spectroscopy studies of incubated eggs indicated no significant difference (P > .05) in the fat:water ratio between the two strains. The difference in the PME:PDE ratios between the two strains as obtained by 31P-NMR spectroscopy over all days of incubation analyzed was not significant (P > .05); however, there was a significant difference in this ratio between the two strains at Days 4, 6, and 11. Up to Day 16, Strain 30 had a slightly, but not significantly (P > .05), higher ATP:PDE ratio as shown on 31P-NMR spectra, whereas after Day 17 the ATP:PDE ratio was significantly higher (P < .01) for Strain 31. Strain 31 birds had a significantly lower (P < .05) crude fat content. There was a significant difference (P < .05) in 7-wk carcass fat content between sexes, males having significantly (P < .01) less fat than females, which was correlated with a significantly higher (P < .01) ATP:PDE ratio in male embryos. It might be possible to use ATP:PDE ratios obtained during embryonic development by 31P-NMR to select strains of birds for low fat content at market weight and to distinguish between sexes during late embryonic development.

Adipose Tissue↗
PubMed1991

Cellular energy metabolism and regulation.

Consistent with the increased demand for nutrients imposed by lactation and growth, those tissues directly involved in the digestion, absorption, and processing of the required additional nutrients show response to these states. During lactation, the rumen, upper intestine, and liver increase in size, and more energy is spent on Na+,K+ transport and on protein turnover. The massive endocrine influences during lactation suggest that the metabolism of other tissues besides these and mammary tissue would be influenced, but evidence is rather sparse. Ion transport and protein metabolism in some muscles may indeed be increased. Although substrate cycles characteristically account for a substantially smaller portion of the energy expenditure in the intact animal than do ion transport and protein turnover, stage of lactation influences some of these cycles, particularly the triacylglycerol fatty acid cycle. The needs for additional quantitative in vivo measurements of metabolic conversions and for mechanistic model description of metabolic events in nonmammary tissues are discussed.

Absorption↗
PubMed1989

A quantitative model of reticulo-rumen particle degradation and passage.

Labelled particles were prepared by mordanting low concentrations (0.1 or 5 g/kg dry matter) of chromium to neutral-detergent-extracted stems (1-2 mm or 10 mm in length) of bromegrass (Bromus inermis). These were used in the study of reticulo-rumen particle kinetics of four steers given bromegrass hay and from the results a quantitative model of particle digestion and passage was developed. At the 0.1 g Cr/kg concentration there was minimal interference with digestibility of the feedstuff. The ratio, dry weight of the reticulo-rumen large-particle pool (greater than 3.35 mm): small-particle pool (less than 3.35 mm) was 2:1. It was derived from the model that volatile fatty acids (VFA) and carbon dioxide in the rumen were produced mainly from large particles, and that between 500 and 700 g/kg hay dry matter was digested in the reticulo-rumen. It was also derived from the model that a major portion, 200 (SE 110) g/kg, of the hay dry matter was rapidly solubilized and that the material leaving the reticulo-rumen was composed of small particles (500-840 g/kg), large particles (100-160 g/kg) and an unknown portion of soluble dry matter of hay (0-400 g/kg). Disappearance from the large-particle pool in the model involving the lowest Cr level was directed to formation of VFA and CO2 (0.68 (SE 0.04) of total flow) to the small-particle pool (0.25 (SE 0.06) of total flow) and direct passage from the reticulo-rumen (0.07 (SE 0.002) of total flow). The disappearance from the small-particle pool was to VFA and CO2 production and to the omasum accounting for 0.14 (SE 0.18) and 0.86 (SE 0.24) respectively, of the total flow. It was concluded that the low-level-mordanting technique in combination with appropriate sampling yielded a realistic quantitative description of forage breakdown and movement processes in the digestive tract of cattle.

Animal Feed↗
PubMed1989

Mathematical integration of protein metabolism in growing lambs.

A mathematical integration of whole-body protein synthesis and degradation based on protein metabolism in 10 individual tissues in growing lambs is described. The tissues represented are adipose, central nervous system (CNS), gastrointestinal tract (GIT), heart, kidney, liver, muscle, pancreatic and salivary glands (PSG), reticuloendothelial system (RES) and skin, together with a blood pool of amino acids. The fluxes represented in the equations are generally assumed to follow simple or modified mass-action kinetics. The fractional rates for protein synthesis in each tissue were calculated from published values for lambs, where these were available, or alternatively derived using assumptions based on data obtained with rats. The initial protein content of each tissue and the fractional degradation rate were assigned values calculated using data from a slaughter experiment. The model was used to examine whole-body protein synthesis at different rates of growth and to compare the relative contributions of each tissue. The GIT (25-26%) and skin (23-26%) had the highest contributions to total protein synthesis, followed by muscle (21-26%), liver (13-14%), RES (6-7%) and PSG (3-6%), while adipose, CNS, heart and kidney together contributed less than 5%. These values agree reasonably well with experimental values, and thus the model can be used to examine the effect of different growth rates on protein metabolism and its associated energy costs.

Amino Acids↗
PubMed1989

Simulation of the energy costs associated with protein turnover and Na+,K+-transport in growing lambs.

A mathematical representation of the energy-requiring processes of protein turnover and Na+,K+-transport in the tissues of growing lambs is described. This model was then used to examine the relative contributions of these processes to ATP expenditure at two different growth rates (90-230 g/d). Protein turnover accounted for 19% of whole-body ATP expenditure at both growth rates examined, with the gastrointestinal tract (GIT), accounting for 25-27%, muscle for 21-26%, skin for 23-26% and liver for 13% of total protein turnover energy costs. The contribution of Na+,K+-transport increased from 18 to 23% of whole-body heat production as growth rate increased, with the GIT accounting for 39 and 50%, muscle for 17 and 10% and liver for 18 and 23% of total Na+,K+-transport costs at low and high nutrient inputs, respectively. Thus, protein turnover accounted for 19% of the increment in ATP expenditure due to the increased nutrient input at the higher rate of growth, while Na+,K+-transport accounted for 39%, and fat turnover and accretion accounted for 25%, leaving 17% of the ATP increment unaccounted for.

Adenosine Triphosphate↗
PubMed1985

Magnitude of ouabain-sensitive respiration of lamb hepatocytes (Ovis aries).

In lamb hepatocyte preparations with viabilities greater than 90%, ouabain-sensitive respiration accounted for approximately 50% of the total cellular O2 consumption. Lamb hepatocyte preparations with viability of less than 50% exhibited lower (P less than 0.05) total and ouabain-sensitive respiration. The decrease in ouabain-sensitive respiration in these preparations entirely accounted for the drop in total respiration.

Age Factors↗
PubMed1985

The effect of lumen conditions on oxygen uptake in perfused omasal laminae.

The vascular anatomy of the bovine omasal lamina permitted perfusion of a discrete area of the tissue. As occurs in vivo, oxygen was provided through the vascular system, while the luminal sides of the tissue could be kept in an anaerobic environment, thus allowing study of foregut tissue metabolism under physiologically realistic conditions. O2 consumption of perfused leaves in the presence of anaerobic buffer was 64.9 and 73.5 nmol O2/mg dry weight per h in Expts 1 and 2 respectively, and was elevated (P less than 0.05) when the lumen side of the tissue was exposed to an atmosphere of nitrogen gas. In Expt 1, the rate of O2 consumption was increased (P less than 0.01) by 35% as a result of suspension of a boiled preparation of rumen micro-organisms and particles (less than 1 mm) in the anaerobic lumen buffer. Replacement of the boiled preparation with an unboiled suspension increased O2 consumption further by 11%, but this was not statistically significant (P greater than 0.05). In Expt 2, sequential addition of the following substrates or preparations to the lumen chambers all resulted in stepwise increases (P less than 0.05) in O2 consumption; 8 mM-butyrate, boiled rumen micro-organisms and particles and, finally, unboiled rumen micro-organisms and particles. Identities of the heat-labile and heat-stable components of the microbial and particle suspensions that caused enhancement of O2 removal across the perfused tissue are discussed.

Animals↗
PubMed1985

Removal of digesta components from the rumen of steers determined by sieving techniques and fluid, particulate and microbial markers.

When 103Ru-labelled Tris (1,10-phenanthroline) ruthenium II chloride (103Ru-P) particulate marker in aqueous solution was added to the rumen of four steers given 5.5 kg grass hay/d at two-hourly intervals, the distribution of 103Ru-P marker among rumen particles of various sizes was the same at 4 h, 3 d and 7 d after administration, the concentration of 103Ru-P/g dry matter (DM) was inversely related to particle size and 0.30 of the 103Ru-P was associated with the DM of particles too large to be moved from the rumen at a meaningful rate. Thus, fractional outflow rate (FOR) of 103Ru-P would reflect, but was not a direct measure of, the FOR of the small particle pool in the rumen. When rumen digesta were labelled with 103Ru-P, placed in nylon cloth bags and incubated in vitro with unlabelled digesta, 59% of the 103Ru-P disappeared from the nylon bag in 24 h, and 74% in 48 h. Similar results were obtained when large particles (retained by a 3.2 mm mesh screen during wet sieving) from rumen digesta were subjected to this procedure. In a further experiment, the steers were given the hay in either the long or ground form and drinking water to which 10 g sodium chloride/l were, or were not, added. The FOR of 51CrEDTA in centrifuged rumen fluid was increased (P less than 0.05) from 1.78 to 2.10/d by grinding of the hay diet, but was not influenced by the intake of an additional 257 g NaCl/d. The FOR values of 103Ru-P in mixed rumen digesta and organic 35S in micro-organisms were linearly correlated (P less than 0.05) and were not affected (P greater than 0.05) by grinding and salt treatments. On average, the FOR of organic 35S in micro-organisms was 0.41 of that of 51CrEDTA in centrifuged rumen fluid and 0.85 of that of 103Ru-P in rumen digesta respectively. Grinding of the hay did not (P greater than 0.05) change the proportion of rumen DM (0.476-0.515) or faecal DM (0.107-0.153) retained by the 3.2 mm mesh and larger screens. FOR from the rumen of a given size group of particles was calculated as the ratio, estimated daily flow from the rumen of the size group: rumen pool of the group.(ABSTRACT TRUNCATED AT 400 WORDS)

Animal Feed↗
PubMed1985

Influence of feed intake and starvation on the magnitude of Na+,K+-ATPase(EC 3.6.1.3)-dependent respiration in duodenal mucosa of sheep.

Oxygen consumption and Na+,K+-ATPase(EC 3.6.1.3)-dependent (ouabain-sensitive) and -independent respiration were measured for duodenal mucosa biopsies from 10-month-old sheep given two levels of digestible energy (DE) intake (7.6-7.7 and 14.8 MJ lucerne (Medicago sativa) pellets/d) and following 48 h of starvation. The mucosal biopsies were determined to be structurally intact and free of adherent bacteria on histological and scanning-electron-microscope examinations. The use of D-glucose as a substrate during incubations did not elevate (P greater than 0.05) the respiration indices of the biopsies over those measured during acetate incubations. Glucose uptake did not (P greater than 0.05) influence the Na+,K+-ATPase-dependent respiration of the mucosal biopsies. Na+,K+-ATPase-dependent respiration accounted for 50% of the total O2 consumption of the mucosal biopsies of sheep given the lower level of DE. Total O2 consumption of the duodenal mucosa was not (P greater than 0.05) increased when sheep were given the higher level of DE but Na+,K+-ATPase-dependent respiration of the mucosa was elevated (P less than 0.01) by 37% during this period. When sheep were starved for 48 h, total O2 consumption of the mucosal biopsies was not (P greater than 0.05) affected, however, Na+,K+-ATPase-dependent respiration of the biopsies dropped (P less than 0.01) by 45%. Na+,K+-ATPase-dependent respiration accounted for 61.3% of the O2 uptakes of mucosa from the sheep given the higher level of DE and 28.3% of the O2 uptake of mucosa from fasted sheep.

Animals↗
PubMed1985

Magnitude of ouabain-sensitive respiration in the liver of growing, lactating and starved sheep.

Oxygen consumption and ouabain-sensitive respiration was measured for liver biopsies from lactating and non-lactating ewes and for hepatocytes isolated from mature, dry ewes. O2 consumption, ouabain-sensitive respiration and 86Rb+ uptake were also measured for hepatocytes isolated from lambs, fed adult sheep and adult sheep starved for 5 d. Ouabain-sensitive respiration in the liver of ewes at peak lactation accounted for 45% of the total liver O2 consumption. This percentage was 24-37% higher (P less than 0.05) than measurements made during late lactation and during the non-lactating period. Total O2 consumption and ouabain-sensitive respiration rates of lamb hepatocytes were greater (P less than 0.05) than similar measurements for hepatocytes isolated from adult sheep. Ouabain-sensitive 86Rb+ uptake by hepatocytes from fed sheep was up to six times greater (P less than 0.05) than that by cells from starved sheep. The magnitude of ouabain-sensitive respiration of hepatocytes from starved sheep was 62% lower (P less than 0.05) than that for hepatocytes from fed sheep.

Animals↗
PubMed1985

Energy costs of ion pumping by animal tissues.

Results from recent in vitro studies indicate that in excess of 20% of the energy expenditure of skeletal muscle, duodenal epithelium and liver of domestic ruminants is to achieve Na+ and K+ transport across the plasma membrane. The energy cost of active Ca2+ transport is less clear but is likely less than 10% of the total expenditure of skeletal muscle at rest. Energy expenditure on Na+ and K+ transport was quite sensitive to the physiological state of the animal. During lactation, Na+ and K+ transport accounted for nearly half of the in vitro O2 uptake of skeletal muscle, duodenal epithelium and liver. The energetic cost of supporting Na+ and K+ transport was also elevated in young, as compared with older animals, by feed intake and by exposure to cold. Na+ and K+ transport appears to be a substantial component of the maintenance energy expenditure of ruminant tissues. Its variation, therefore, implies that change of maintenance energy expenditures with physiological state of the animal warrants serious attention.

Adenosine Triphosphate↗
PubMed1984

A comparison of methods for the estimation of the proportion of microbial nitrogen in duodenal digesta, and of correction for microbial contamination in nylon bags incubated in the rumen of sheep.

Four sheep, each fitted with cannulas in the rumen and proximal duodenum, were given two diets (1390 g dry matter (DM)/d) consisting of lucerne (Medicago sativa) pellets (24.2 g nitrogen/kg DM) plus pelleted reed canary grass (Phalaris arundinacea; 14.1 g N/kg DM) or chopped hay (11.8 g N/kg DM) at intervals of 2 h. Flow of duodenal digesta measured by reference to the markers 51Cr-EDTA and 103Ru-phenanthroline indicated a net gain of 5.8-7.5 g non-ammonia-N (NAN) between mouth and duodenum. The proportion of microbial N in duodenal digesta N was estimated using 15N and 35S incorporation into bacteria and digesta. Two methods of analysis for 35S content, the Bird & Fountain (1970; B&F method) and the Mathers & Miller (1980; M&M method), were used. (15NH4)2SO4 and Na2(35)SO4 were infused into the rumen for 3.5 d before and 4.0 d during sampling. A bacterial fraction was prepared from the fluid phases of sampled duodenal digesta and rumen contents by differential centrifugation. In addition, samples of ground canary grass and of lucerne were incubated in nylon bags in the rumen for 3-48 h during the infusion. Each of the 35S analytical methods yielded similar values of 35S content of isolated rumen or duodenal bacteria, but there was more (P less than 0.05) incorporation of 15N into rumen than into duodenal bacteria. Relative to values obtained using the M&M method and 15N incorporation, the B&F method for S analysis yielded higher (P less than 0.05) estimates of microbial content of duodenal digesta from sheep given chopped reed canary grass. 35S activity associated with washed nylon-bag residues increased rapidly with time-period of incubation and was substantially greater (P less than 0.05) when analysed by the B&F method compared with the M&M method. The 35S content (/g DM) of adherent bacteria removed from nylon-bag residues by homogenization in a second experiment varied from 0.65 to 1.88 that of free-living bacteria isolated from rumen fluid by differential centrifugation. The difference in 35S content in digesta and nylon-bag residues as measured using the two analytical methods was considered in relation to 35S-labelled extracellular material postulated to be produced by bacteria adherent to plant residues. Estimates of disappearance of dietary N from nylon bags after correction for microbial contamination indicated a disparity with estimates based on in vivo information.

Animal Feed↗
PubMed1983

In vitro degradation of leucine in muscle, adipose tissue, liver, and kidney of fed and starved sheep.

In vitro rates of conversion of [1-14C]leucine to 4-methyl-2-oxo[1-14C]pentanoate and of oxidation of [1-14C] and [U-14C]leucine were measured for tissues from fed and starved (5 days) sheep. Slices of liver and kidney and preparations of adipose tissue and of fibre bundles of external intercostal muscle (EIC) were used. Skeletal muscle is likely the major site of leucine catabolism in sheep although adipose tissue is capable of substantial metabolism. Muscle and adipose tissue from fed sheep released 17 and 5% of the [1-14C]leucine transaminated as 4-methyl-2-oxo-[1-14C]pentanoate and upon starvation the proportions were increased (P less than 0.001) to 46 and 32%. Starvation reduced (P less than 0.01) leucine catabolism in all tissues except the kidney. The pattern of leucine catabolism in EIC muscle changed from extensive oxidation in the fed state to being limited essentially to transamination and decarboxylation in the starved state.

Adipose Tissue↗
PubMed1983

Simultaneous determination of total cholesterol concentration and radioactivity in plasma.

Total plasma cholesterol concentration and radioactivity were measured simultaneously using a gas chromatograph equipped with a flame ionization detector and an effluent splitter. More than 99% of the recovered radioactivity was in the cholesterol peak. Specific activities were highly correlated with the amounts of labeled cholesterol present in plasma. The recovery of label was quantitative over a wide range of carrier cholesterol concentrations. The method is highly reproducible, accurate, rapid and specific.

Acetates↗