Combined effects of diet and cold exposure on insulin responsiveness to glucose and tissue responsiveness to insulin in sheep.

Hyperglycemic clamp and hyperinsulinemic euglycemic clamp techniques were used to investigate effects of diet and cold exposure on insulin responsiveness to glucose and tissue responsiveness and sensitivity to insulin in adult rams. The sheep were fed a high-concentrate diet (80% concentrate and 20% roughage) and a high-roughage diet (20% concentrate and 80% roughage), both 70% above the ME requirement for maintenance, and were exposed to a thermoneutral environment (20 degrees C) and a cold environment (0 degrees C) for 2 wk. The estimated ME intake was greater (P < .01) for the high-concentrate diet than for the high-roughage diet. In the hyperglycemic clamp experiment, the ratio of the plasma insulin increment to the glucose infusion rate (insulin responsiveness to glucose) was lower during cold exposure than in the thermoneutral environment in sheep fed the high-concentrate diet but was unchanged in sheep fed the high-roughage diet. In the hyperinsulinemic euglycemic clamp experiment, the glucose infusion rate (tissue responsiveness to insulin) was higher (P < .01) for the high-concentrate diet than for the high-roughage diet, and it was also higher (P < .01) during cold exposure than in the thermoneutral environment, indicating that tissue responsiveness to insulin was intensified in sheep fed the high-concentrate diet during cold exposure associated with the higher energy intake. These results suggest that both reduced insulin responsiveness and enhanced tissue responsiveness to insulin in sheep exposed to cold were dependent on the type of diet.     

Chromium supplementation does not influence glucose metabolism or insulin action in response to cold exposure in mature sheep

An isotope dilution method using [U-(13)C] glucose infusion and a glucose clamp approach were applied to determine the effects of supplemental Cr and cold exposure on blood glucose turnover rate and tissue responsiveness and sensitivity to insulin in eight sheep. The daily profiles of blood metabolites and hormones were also determined. The sheep consumed diets containing either 0 or 1 mg of Cr/kg from a high-Cr yeast and were exposed from a thermoneutral environment (20 degrees C) to a cold environment (0 to 4 degrees C) for 9 d. The experiment used a crossover design. Body weight was lost (P = 0.02) during cold exposure, regardless of Cr supplementation. Blood glucose turnover rate and the maximal glucose infusion rate did not differ between diets, but both were higher (P = 0.0004 and P = 0.0001, respectively) during cold exposure than in the thermoneutral environment. The plasma insulin concentration at half-maximal glucose infusion rate changed with neither diet nor environment. Plasma concentrations of glucose and NEFA increased (P < 0.05) during cold exposure for both diets. In sheep, Cr supplementation, 1 mg/kg of diet as high-Cr yeast, has little influence on blood glucose metabolism and insulin action, whereas cold exposure enhances both without further modification by Cr supplementation.     

Effect of hypomagnesemia and cold exposure on tissue responsiveness to insulin in sheep given a low magnesium and high potassium diet

Hypomagnesemia in ruminants has been shown to be associated with cold stress and with altered insulin secretion and sensitivity. However, the relationship between hypomagnesemia and tissue responsiveness to insulin in ruminants exposed to cold environment is still unclear. The hyperinsulinemic euglycemic clamps, four insulin infusion rates (1, 2, 4 and 8 mU x kgBW(-1) x min(-1) for four sequential periods of 2-h each) were performed to determine combined effects of hypomagnesemia and cold exposure (0 degrees C) on tissue responsiveness to insulin in sheep. The low magnesium (0.05% Mg) and high potassium (4.20% K) diet and cold exposure decreased (P <.05) plasma Mg levels compared with those of the control diet (0.28% Mg/0.52% K) and the thermoneutral environment (20 degrees C). In the euglycemic clamps, cold exposure increased (P <.01) the pooled glucose infusion rate (GIR) across various insulin infusion rates in both diet treatments, though the increases in GIR were small for the low Mg/high K diet. In the cold environment, the GIR was lower (P <.01) for the low Mg/high K diet-fed sheep than for the control diet-fed sheep. The maximal insulin-induced increase in GIR was lower (P <.01) in the hypomagnesemic sheep than in the control sheep during cold exposure, and the insulin level resulting in half-maximal GIR tended to be higher in the hypomagnesemic sheep than in the control sheep. These results indicated that hypomagnesemia in ruminants depressed the enhanced tissue responsiveness to insulin in the cold environment, and decreased insulin-mediated glucose disposal.     

Effects of feed restriction and cold exposure on glucose metabolism in response to feeding and insulin in sheep.

The effects of feed restriction, cold exposure, and the initiation of feeding on blood glucose metabolism, other blood metabolites, hormones, and tissue responsiveness and sensitivity to insulin were measured in sheep. The sheep consumed orchardgrass hay ad libitum (AL) or were restricted to 82% of the ME requirement for maintenance (RE) and were exposed to a thermoneutral (20 degrees C) or a cold environment (2 degrees C). An isotope dilution method and a glucose clamp approach were applied to determine blood glucose metabolism and insulin action, respectively. Plasma NEFA and insulin concentrations were influenced by feed restriction. Concentrations of plasma glucose, NEFA, insulin, and glucagon were influenced by cold exposure. Plasma NEFA concentration for RE decreased after the initiation of feeding and plasma insulin concentration increased transiently for all treatments. [U-13C]Glucose was continuously infused for 8 or 7 h after a priming injection starting 3 h before the initiation of either feeding or insulin infusion, respectively. When responses to feeding were studied, blood glucose turnover rate was less (P < .001) for RE than for AL, and it was greater (P < .001) during cold exposure than in the thermoneutral environment. The rate changed little after the initiation of feeding. For the glucose clamp approach, insulin was infused over four sequential 1-h periods at rates from .64 to 10 mU x kg BW(-1) x min(-1), with concomitant glucose infusion to maintain preinfusion plasma glucose concentrations. The rates of glucose infusion and blood glucose turnover increased (P < .001) dose-dependently with insulin infusion rate. The maximal glucose infusion rate was greater (P < .05) for RE than for AL and was greater (P < .001) during cold exposure than in the thermoneutral environment. The plasma insulin concentration at half-maximal glucose infusion rate was lower (P < .1) during cold exposure. Blood glucose turnover rate tended to be greater (P = .10) for RE than for AL, and it was greater (P < .001) during cold exposure than in the thermoneutral environment. The ratio of endogenous production to utilization of glucose was suppressed by insulin infusion. In sheep fed a roughage diet, blood glucose turnover rate seems to be influenced by both intake level and environmental temperature, but not by the act of feeding. Moreover, the action of insulin on glucose metabolism is enhanced during cold exposure, and the effect of feed restriction is somewhat enhanced.     

Effects of dietary energy intake and cold exposure on kinetics of plasma glucose metabolism in sheep

An isotope dilution method using [U-(13)C]glucose injection was applied to determine the effects of dietary energy intake and cold exposure on plasma glucose metabolism in sheep. The sheep were assigned to two dietary treatments and were fed on diets containing either 100% or 160% of ME and both containing 150% of dietary crude protein intake for maintenance. The sheep were exposed from a thermoneutral environment (23 degrees C) to a cold environment (2-4 degrees C) for 5 days. The isotope dilution method was performed on the 18th day in the thermoneutral environment and on the fifth day of cold exposure. Plasma concentrations of glucose and NEFA increased (p < 0.05) during cold exposure for both diets. Plasma glucose pool size remained unchanged (p = 0.67), but plasma glucose turnover rate tended to increase (p = 0.07) with increased energy intake. Both pool size and turnover rate of plasma glucose increased (p = 0.01 and p = 0.0001, respectively) during cold exposure. No significant diet x environment interaction was detected. It is concluded that plasma glucose metabolism was influenced by both dietary energy intake and cold exposure, and plasma glucose metabolism in response to cold exposure was not modified by energy intake in sheep under the conditions (2-4 degrees C on the fifth day) of the present experiment.     

Acetyl-CoA carboxylase and fatty acid synthase activity and immunodetectable protein in adipose tissues of ruminants: effect of temperature and feeding level

To gain insights into the regulation of fat synthesis, we have investigated the effect of cold environmental exposure and feed restriction of sheep on activity and immunodetectable protein content of acetyl-CoA carboxylase (ACC) and fatty acid synthase in adipose tissue. Subcutaneous and mesenteric adipose tissues were collected at slaughter from sheep exposed to either cold (0+/-2 degrees C) or warm (23+/-2 degrees C) environment, and given either ad libitum or restricted access to feed for three 5-wk periods. Acetyl-CoA carboxylase was isolated from frozen adipose tissue samples and activity determined as the rate of incorporation of H14CO3- into acid stable malonyl-CoA. Cold exposure and feed restriction reduced (P < .05) ACC activity in the two adipose tissue depots. Western blot analysis with peroxidase-conjugated streptavidin showed that both adipose tissue depots express a single isoform of ACC. In s.c. adipose tissue, cold exposure increased (P < .05) ACC protein abundance, which is opposite to the change in activity. However, feed restriction reduced immunodetectable ACC protein. There was no significant effect of environment or feeding level on ACC protein abundance in mesenteric tissue. Fatty acid synthase activity determined in ammonium sulfate extract by measuring the malonyl-CoA- and acetyl-CoA-dependent oxidation of NADPH was decreased (P < .05) by feed restriction in both s.c. and mesenteric tissues. Cold exposure reduced fatty acid synthase activity in s.c. but not in mesenteric tissue. There was no effect of environment on fatty acid synthase protein abundance in either adipose tissue depot. However, feed restriction significantly reduced fatty acid synthase protein abundance in the two depots. The data suggest that feed restriction and exposure of ruminants to cold environmental conditions may significantly down-regulate the activity of key lipogenic enzymes.     

Insulin responsiveness to glucose and tissue responsiveness to insulin over the feeding cycle in sheep

Insulin responsiveness to glucose and tissue responsiveness to insulin, using the hyperglycemic clamp and the hyperinsulinemic euglycemic clamp techniques, were measured before, during and after feeding in sheep fed an alfalfa hay and commercial concentrate diet. Glucose infusion rate and the plasma insulin increment in the hyperglycemic clamp experiment were higher during the feeding period (0 to 1 h after initiating feeding) than during the pre- and post-feeding periods. The ratio of plasma insulin increment to glucose infusion rate remained unchanged over the feeding cycle. Only a slight increase (P less than .05) in the glucose infusion rate was observed during feeding in the hyperinsulinemic euglycemic clamp experiment. These results suggest that insulin responsiveness to glucose tends to be enhanced during the feeding period but that tissue responsiveness to insulin is not changed over the feeding cycle in sheep.     

Effects of exogenous glucose or colostrum on body temperature, plasma glucose, and serum insulin in cold-stressed, newborn Brahman calves.

The effects of ambient temperature and source of exogenous energy (glucose or colostrum) on the ability of newborn Brahman calves to maintain rectal temperature (RT) were determined. All calves were removed from dams within 30 min of birth, before suckling. Calves were catheterized and placed in either a warm (25 degrees C) or cold (5 degrees C) environment for 150 min and given either colostrum or glucose. This resulted in four groups (warm colostrum, n = 7; cold colostrum, n = 7; warm glucose, n = 6; cold glucose, n = 6). Blood samples and RT were obtained at 15-min intervals during warm or cold through 150 min, when calves were removed from cold, and at 180, 240, and 300 min. After 60 min, each calf was given either 1 L of colostrum (38 degrees C) from its dam or glucose (38 degrees C) infusion of 750 mg/kg BW. Plasma glucose concentrations were determined by enzymatic techniques and serum insulin concentrations by RIA. Calves exposed to cold or warm air temperatures had similar declines in rectal temperature from 0 to 60 min. Colostrum-fed, cold calves had a greater (P less than .07) decrease in RT than did colostrum-fed, warm calves from 75 through 150 min; glucose-infused warm and cold calves had intermediate decreases in RT. Plasma glucose increased (P less than .0001) in glucose-infused compared with colostrum-fed calves at 75 min, but glucose-infused calves had lower (P less than .02) glucose levels from 180 to 300 min. Higher (P less than .05) glucose concentrations     

不同精粗比饲粮中添加甘露寡糖对绵羊瘤胃养分降解率的影响

本试验采用尼龙袋法研究不同精粗比饲粮中添加甘露寡糖(MOS)对绵羊瘤胃养分降解率的影响。选取6只安装永久瘤胃瘘管的羯羊(白萨福克×小尾寒羊♀),采用4×6双因子析因试验设计,共设饲粮精粗比和MOS添加水平2个因子,其中饲粮精粗比分别为20∶80(A1)、30∶70(A2)、40∶60(A3)和50∶50(A4),MOS添加水平分别为0(B1)、0.4%(B2)、0.8%(B3)、1.2%(B4)、1.6%(B5)和2.0%(B6)。结果显示:饲粮精粗比对不同时间点绵羊瘤胃干物质(DM)、有机物(OM)和粗蛋白质(CP)的降解率、降解参数[DM、OM:快速降解部分(a)、慢速降解部分(b)和可降解部分(a+b);CP:a、b和b的降解速率(c)]及有效降解率均产生了显著影响(P0.05),其变化规律均为随饲粮精料比例的升高而升高,在A4组达到最高;MOS添加水平仅对不同时间点绵羊瘤胃CP的降解率及降解参数a+b产生了显著影响(P0.05),其变化规律均为随MOS添加水平的增加先升高后降低,在B5组达到最高。饲粮精粗比和MOS添加水平对绵羊瘤胃养分降解率没有产生显著的交互作用(P0.05)。由此得出,饲粮精粗比为50∶50时绵羊瘤胃干物质、有机物和粗蛋白质的有效降解率最高,MOS添加水平达到1.6%时绵羊瘤胃粗蛋白质的降解率最高。     

Insulin responsiveness to glucose and tissue responsiveness to insulin in lactating, pregnant, and nonpregnant, nonlactating beef cows

Insulin responsiveness to glucose and tissue responsiveness to insulin, using the hyperglycemic clamp and the hyperinsulinemic euglycemic clamp techniques, were measured in lactating, late pregnant, and nonpregnant, nonlactating (NPNL) beef cows. The glucose infusion rate (GIR) in the hyperglycemic clamp technique was higher (P less than .05). in lactating cows than in NPNL cows. The plateau in plasma insulin concentration (insulin responsiveness) was higher (P less than .05) in lactating cows than in late pregnant and NPNL cows. Pregnant cows tended to have higher GIR and lower plateau in plasma insulin concentration than NPNL cows. In the hyperinsulinemic euglycemic clamp technique, GIR (tissue responsiveness to insulin) was higher (P less than .05) in lactating cows than in late pregnant cows; values for NPNL cows were intermediate. We conclude that insulin responsiveness to glucose and tissue responsiveness to insulin were enhanced during lactation but tended to be decreased during late pregnancy in beef cows.     

Muscle growth and plasma concentrations of amino acids, insulin-like growth factor-I, and insulin in growing pigs fed reduced-protein diets

Twenty barrows were used to determine if partial replacement of protein-bound AA with crystalline AA (CAA) reduces AA use for muscle tissue and whole-body growth. Barrows (44.2 +/- 1.3 kg of BW) were assigned to 4 diets in a randomized complete block design. Diets consisted of 16.1% CP with no CAA, and 12.8, 10.1, and 7.8% CP containing CAA. As the CP concentration decreased, CAA were gradually increased to meet requirements on a true ileal digestibility basis. Barrows were weighed on d 0 and 13. Blood samples were collected before the morning feeding on d 0, 6, and 12 (prefeeding), and 2 h after the morning feeding on d 13 (postfeeding). Pigs were euthanized on d 13, and liver and right LM were removed and weighed. The reduction in the dietary CP concentration linearly decreased (P < 0.01) ADG, G:F, LM weight, and the CP content of LM. Reducing the CP concentration decreased pre- and postfeeding plasma concentrations of IGF-I (linear, P < 0.01) and insulin (linear, P < 0.10). The reduction in the dietary CP concentration increased prefeeding plasma concentrations of Ala, Gln, Gly, and total AA but decreased Arg, Asn, His, Ile, Phe, Trp, and Tyr (linear, P < 0.05). Plasma concentration of total indispensable AA decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.01). The reduction in the dietary CP concentration increased postfeeding plasma concentrations of Ala, Lys, Met (linear, P < 0.01), and Gly (linear, P = 0.073) and decreased Asn, Ser, Tyr, Arg, His, and Leu (linear, P < 0.05). Plasma concentrations of Ile, Phe, Thr, Trp, and Val decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.05). In muscle tissue, concentrations of free Ala, Asp, Glu, Gln, Gly, and Lys increased (linear, P < 0.05) as the dietary CP concentration decreased. Concentrations of free His, Ile, Phe, Thr, Trp, and Val in muscle tissue decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.05). In summary, the reduction in the dietary protein-bound AA decreased whole-body and LM growth, altered the free AA pool profile in muscle tissue, and decreased plasma insulin and IGF-I. As the replacement of protein-bound AA with CAA increased, 1) free Ala and Gln in muscle tissue increased, indicating an increase of muscle tissue protein breakdown; and 2) utilization of indispensable AA in muscle tissue decreased.     

Effects of oscillating dietary protein on ruminal fermentation and site and extent of nutrient digestion in sheep

Eight cannulated wethers (BW = 52.5 +/- 5.7 kg) were used in a replicated 4 x 4 Latin square designed experiment to evaluate the effects of oscillating dietary protein concentrations on ruminal fermentation, site and extent of digestion, and serum metabolite concentrations. Four treatments consisted of a 13, 15, or 17% CP diet fed daily or a regimen in which dietary CP was oscillated between 13 and 17% on a 48-h basis (ACP). All diets consisted of 65% bromegrass hay (10.5% CP, 61.9% NDF, 37.2% ADF) plus 35% corn-based supplement and were formulated to contain the same amount of degradable intake protein (9.6% of DM) plus additional undegradable intake protein (SoyPLUS, West Central Cooperative, Ralston, IA) to accomplish CP levels above 13%. Each of four experimental periods were 16 d in duration with 12 d for diet adaptation followed by 4 d for sample collection. All wethers were fed at 3.0% of initial BW (DM basis) throughout the experiment, resulting in an average organic matter intake of 1.39 kg/d across treatments. When compared to the 15% CP daily treatment, feeding ACP had no effect (P > or = 0.10) on ruminal or lower tract N, NDF, ADF, or OM digestion. True ruminal OM digestion responded quadratically (P = 0.07) to increasing dietary CP, reaching a maximum of 52.0% of OM intake with the 15% CP treatment. Sheep fed ACP tended to have lower (P = 0.08) ruminal NH3 N concentrations and an overall higher (P = 0.0001) molar proportion of acetate compared to those fed 15% CP daily. Total VFA concentrations were not affected (P > or = 0.45) by increasing dietary CP. Microbial efficiency did not differ (P > or = 0.55); thus, bacterial N flow at the duodenum responded quadratically (P = 0.04) to increasing dietary CP. Nonbacterial N (P = 0.001) and total N (P = 0.01) flows at the duodenum and total tract N digestibility (P < or = 0.04) increased linearly as dietary CP increased. Wethers fed ACP maintained a lower (P = 0.002) serum glucose and lower (P = 0.0006) serum urea N compared to those fed 15% CP daily. Because the CP content of the diet was increased at the expense of corn, the response to increased CP observed in this experiment is most likely due to negative associative effects of supplemental starch on ruminal fermentation and microbial growth. Oscillating the CP content of the diet on a 48-h basis has little effect on digestion or N utilization in sheep compared with feeding the same quantity of protein on a daily basis.     

Effects of ammonia load on glucose metabolism by isolated ovine duodenal mucosa

To determine the effects of ammonia load on glucose metabolism in ruminant small intestinal tissues, duodenal mucosal cells (DMC) were isolated from growing female sheep (n = 10; 46. 0 +/- 0. 8 kg of BW) fed diets differing in CP content: high (19. 4%) vs. low (13. 1%). Ammonia concentration in the duodenal digesta fluid was greater for sheep fed a high CP diet compared with those fed a low CP diet (16. 4 +/- 1. 0 vs. 9. 1 +/- 1. 8 mM). The isolated primary mucosal cells were incubated for 90 min with [2-(13)C] glucose (3 mM) and ammonium chloride (0, 0. 1, 1, 5, 10, 20, or 50 mM) in Krebs-Ringer HEPES buffer. It was hypothesized that DMC would increase glucose carbon utilization for the synthesis of nonessential AA when the ammonia concentration in the incubation media increased. However, utilization of glucose carbon for alanine synthesis decreased linearly (P = 0. 03) as the ammonia concentration in the incubation media increased. Furthermore, glucose disappearance and utilization of glucose carbon for aspartate synthesis were not affected (P > 0. 47) by the ammonia concentration. Contrarily, in vitro glucose disappearance was greater (P = 0. 03) for DMC isolated from sheep fed a low CP diet vs. a high CP diet [14. 6 +/- 1. 6 vs. 8. 6 +/- 1. 3 nmol.(10(6) cells)(-1).(90 min) (-1)], and hexokinase activity was greater (P = 0. 01) in the mucosa of sheep fed a low CP diet compared with a high CP diet (1. 22 +/- 0. 05 vs. 1. 04 +/- 0. 02 mUnit/mg of protein). These observations indicate that ammonia load does not affect the extent of glucose utilization by DMC, and that glucose carbon may not play a significant role for the synthesis of alanine, aspartate, or glutamate when DMC are exposed to increased concentrations of ammonia.     

Porcine somatotropin alters insulin response in growing pigs by reducing insulin sensitivity rather than changing responsiveness

Exogenous porcine somatotropin (pST) treatment consistently improves growth performance and reduces fat deposition in pigs, and it is hypothesized that one component of the mechanism is through altering the sensitivity and/or responsiveness to insulin. Therefore, a study was conducted to investigate the effect of pST treatment on whole-body glucose metabolism in response to varying doses of insulin. Eight barrows were surgically prepared with indwelling catheters and randomly assigned to one of two treatment groups (0 or 120 μg pST/kg BW · d) for 13 d. Whole-body glucose kinetics were measured during infusion of [6-(3)H]-glucose under basal conditions and during hyperinsulinemic-euglycemic clamps at various insulin infusion rates (7, 28, and 140, and 14, 70, and 280 ng insulin/kg BW · min) and alterations in the dose-response parameters were calculated with nonlinear regression. Treatment with pST increased basal plasma concentrations of glucose (36%; P = 0.005), insulin (276%; P = 0.001), and NEFAs (177%; P = 0.01) and decreased the rate of glucose disappearance (-59%; P = 0.001). The responsiveness (maximum response) for steady state glucose infusion rate to maintain glycemia was not altered by pST (112 vs 106 μmol/min · kg; P = 0.78), whereas the sensitivity (effective dose at 50% of maximum response) was increased almost 7-fold (1.3 vs 8.7 ng/mL; P = 0.027). Similar responses were observed for rate of glucose disappearance and insulin-dependent glucose utilization. Therefore, pST-induced insulin resistance with regard to whole-body glucose uptake is due to a reduced sensitivity to insulin, rather than a change in responsiveness.     

Influence of dietary protein level, amino acid supplementation, and dietary energy levels on growing-finishing pig performance and carcass composition

Two experiments were conducted to determine the effects of feeding reduced-CP, AA-supplemented diets at two ambient temperatures (Exp. 1) or three levels of dietary NE (Exp. 2) on pig performance and carcass composition. In Exp. 1, 240 mixed-sex pigs were used to test whether projected differences in heat increment associated with diet composition affect pig performance. There were 10 replications of each treatment with four pigs per pen. For the 28-d trial, average initial and final BW were 28.7 kg and 47.5 kg, respectively. Pigs were maintained in a thermoneutral (23 degrees C) or heat-stressed (33 degrees C) environment and fed a 16% CP diet, a 12% CP diet, or a 12% CP diet supplemented with crystalline Lys, Trp, and Thr (on an as-fed basis). Pigs gained at similar rates when fed the 16% CP diet or the 12% CP diet supplemented with Lys, Trp, and Thr (P > 0.10). Pigs fed the 12% CP, AA-supplemented diet had a gain:feed similar to pigs fed the 16% CP diet when housed in the 23 degrees C environment but had a lower gain:feed in the 33 degrees C environment (diet x temperature, P < 0.01). In Exp. 2, 702 gilts were allotted to six treatments with nine replicates per treatment. Average initial and final BW were 25.3 and 109.7 kg, respectively. Gilts were fed two levels of CP (high CP with minimal crystalline AA supplementation or low CP with supplementation of Lys, Trp, Thr, and Met) and three levels of NE (high, medium, or low) in a 2 x 3 factorial arrangement. A four-phase feeding program was used, with diets containing apparent digestible Lys levels of 0.96, 0.75, 0.60, and 0.48% switched at a pig BW of 41.0, 58.8, and 82.3 kg, respectively. Pigs fed the low-CP, AA-supplemented diets had rates of growth and feed intake similar to pigs fed the high-CP diets. Dietary NE interacted with CP level for gain:feed (P < 0.06). A decrease in dietary NE from the highest NE level decreased gain:feed in pigs fed the high-CP diet; however, gain:feed declined in pigs fed the low-CP, AA-supplemented diet only when dietary NE was decreased to the lowest level. There was a slight reduction in longissimus area in pigs fed the low-CP diets (P < 0.08), but other estimates of carcass muscle did not differ (P > 0.10). These data suggest that pigs fed low-CP, AA-supplemented diets have performance and carcass characteristics similar to pigs fed higher levels of CP and that alterations in dietary NE do not have a discernible effect on pig performance or carcass composition.     

Porcine leptin inhibits lipogenesis in porcine adipocytes

The present study examined whether recombinant porcine leptin alters lipid synthesis in porcine adipocytes. The stromal-vascular cell fraction of neonatal pig subcutaneous adipose tissue was isolated by collagenase digestion, filtration, and subsequent centrifugation. These cells were seeded on 25-cm2 tissue culture flasks and proliferated to confluency in 10% (vol/vol) fetal bovine serum in Dulbecco's modified Eagle medium/F12 (DMEM/F12, 50:50). Cultures were differentiated using 2.5% pig serum (vol/vol), 10 nM insulin, 100 nM hydrocortisone. After 7 d of lipid filling, cultures were washed free of this medium, incubated overnight in DMEM/F12 containing 2% pig serum (vol/vol), and then used for experiments. Acute experiments assessed U-(14)C-glucose or 1-(14)C-palmitate metabolism in cultures exposed to porcine leptin (0 to 1,000 ng/mL medium) for 4 h. Chronic experiments used cultures incubated with 0 to 1,000 ng porcine leptin/mL medium for 44 h before measurements of U-(14)C-glucose and 1-(14)C-palmitate oxidation and incorporation into lipid. Another experiment examined whether chronic leptin treatment alters insulin responsiveness by including insulin (10 nM) with incubations containing leptin. Leptin had no acute effects on glucose oxidation or conversion to lipid (P > 0.05). Acute leptin treatment decreased palmitate incorporation into lipids up to 45% (P < 0.05). Chronic leptin exposure decreased glucose oxidation (21%), total lipid synthesis (18%), and fatty acid synthesis (23%) at 100 ng/mL medium (P < 0.05). Insulin increased rates of glucose oxidation, total lipid, and fatty acid synthesis (P < 0.05); however, chronic exposure to 10 ng leptin/mL medium decreased the effectiveness of 10 nM insulin to affect these measures of glucose metabolism by approximately 18 to 46% (P < 0.05). Higher concentrations of leptin inhibited all effects of insulin on glucose metabolism (P < 0.05). Chronic exposure to leptin increased palmitate oxidation by 36% (P < 0.05). Chronic leptin exposure decreased palmitate incorporation into total lipids by 40% at 100 ng/mL medium (P < 0.05). Lipoprotein lipase activity was not affected (P > 0.05) by leptin. These data indicate that leptin functions to promote partitioning of energy away from lipid accretion within porcine adipose tissue by inhibiting glucose oxidation and lipogenesis indirectly, by decreasing insulin-mediated stimulation of lipogenesis, and by stimulating fatty acid oxidation while inhibiting fatty acid esterification.     

Relationships between plasma concentrations of leptin and other metabolic hormones in GH-transgenic sheep infused with glucose

To study the regulation of leptin secretion in sheep, we infused glucose (0.32 g/h/kg for 12 h) into GH-transgenic animals (n = 8) that have chronically high plasma concentrations of ovine GH and insulin, but low body condition and low plasma leptin concentrations, and compared the responses with those in controls (n = 8). In both groups, the infusion increased plasma concentrations of glucose and insulin within 1 h and maintained high levels throughout the infusion period (P < 0.0001). Compared with controls, GH-transgenics had higher concentrations of insulin, IGF-1, GH (all P < 0.0001) and cortisol (P < 0.05), but lower GH pulse frequency (P < 0.0001). Overall, leptin concentrations were lower in GH-transgenics than in controls (P < 0.01). A postprandial increase in leptin concentrations was observed in both groups, independently of glucose treatment, after which the values remained elevated in animals infused with glucose, but returned to basal levels in those infused with saline, independently of transgene status. In both GH-transgenics and controls, glucose infusion did not affect the concentrations of GH, IGF-1, or cortisol. In conclusion, GH-transgenic and control sheep show similar responses to glucose infusion for leptin and other metabolic hormones, despite differences between them in body condition and basal levels of these hormones. Glucose, insulin, GH, IGF-1 and cortisol are probably not major factors in the acute control of leptin secretion in sheep, although sustained high concentrations of GH and IGF-1 might reduce adipose tissue mass or inhibit leptin gene expression.     

Effects of temperature and plane of nutrition on beta-adrenergic receptors in heart, kidney, and liver of lambs

We determined the effects of temperature and feed intake on beta-adrenergic receptors (beta-adrenoceptors) in tissues of sheep. Twenty-four lambs were exposed during three 5-wk periods to either thermoneutral, control (W; 23+/-2 degrees C) or cold (C; 0+/-2 degrees C) temperatures and were fed either ad libitum (A) or restricted (R) levels of feed intake, resulting in four treatment groups: WA, WR, CA, and CR. Hearts, kidneys, and livers were harvested at slaughter and binding of [3H]dihydroalprenolol to plasma membrane extracts was used to determine densities (B(MAX)) and binding affinities (Kd) of beta1 and beta2 adrenoceptors. The B(MAX) values ranged from 12.10 to 201.26 and 3.38 to 12.30 fmol/mg protein for beta1 and beta2 adrenoceptors, respectively; heart and kidney had the highest and lowest values, respectively. Feed restriction reduced (P < .05) beta1 and beta2 receptor densities in heart but increased (P < .05) beta1 receptor density in kidney and liver. Cold temperature exposure reduced beta1 receptor density in heart tissue during feed restriction. The Kd values, ranging from 1.32 to 5.98 nM, were increased (P < .05) by cold exposure and feed restriction in kidney and liver. Because the effectiveness of hormones is a function of their concentrations, binding affinities, and their receptor densities, these results imply that cold temperature exposure and feed restriction could potentially reduce (in heart) and increase (in kidney and liver) metabolic responsiveness of tissues to catecholamines.     

Pancreatic insulin secretory response and insulin action in heat-exposed sheep given a concentrate or roughage diet

The effects of heat exposure and type of diet on the insulin secretory response to glucose and glucose disposal in response to insulin action in female sheep were investigated employing hyperglycemic and euglycemic clamp techniques. Animals were divided into concentrate and roughage diet groups, and were maintained at the same intake levels of metabolizable energy and crude protein in both diets. Each diet group was subjected to either thermoneutral (20°C, 70% RH) or hot (30°C, 70% RH) environment, followed by glucose clamp experiments.

Heat-exposed sheep showed significant increases in respiration rates (P<.001) and rectal temperature (P<.05). Plasma glucose concentrations in the basal conditions were lower (P<.01) in the hot environment than in the thermoneutral environment, but there was no significant difference in basal levels of plasma insulin between the environmental treatments. In the hyperglycemic clamp experiment, mean plasma insulin increments increased (P<.05) during the heat exposure period across diet treatments. The ratio of mean plasma insulin increment to glucose infusion rate tended to be higher (P<.07) in the hot environment than in the thermoneutral environment, but diet treatment did not affect the ratio of mean plasma insulin increment to glucose infusion rate. The euglycemic clamp technique showed that glucose infusion rates remained unchanged among treatments. Insulin secretion response to glucose could be stimulated in the hot environment.     

Effect of recombinant porcine somatotropin and dietary protein on pancreatic digestive enzymes in the pig.

This study was conducted to determine the effect of daily recombinant porcine somatotropin (rpST) injection (0 or 120 micrograms/kg BW) and dietary CP level of the feed (14 or 26% CP) on pancreatic characteristics of growing pigs. Daily injection of rpST did not affect pancreatic weight (P = .885) but did decrease pancreatic amylase content (P = .005). The ratios of amylase:protein and amylase:trypsin were also lowered by daily rpST injection (P = .002 and P = .0002, respectively). There were protein x rpST and protein x rpST x sex interactions for the ratio of amylase:chymotrypsin. The CP content of the diet had a greater effect than the injection of rpST on pancreatic characteristics. Pigs consuming the 26% CP diet had significantly higher pancreatic weight (P = .003) and greater total pancreatic chymotrypsin (P = .006) than pigs consuming the 14% CP diet. The ratios of trypsin and chymotrypsin to DNA were also higher in pigs fed the 26% CP diet (P = .007 and P = .005, respectively). These responses were not influenced by sex. Recombinant porcine somatotropin seemed to have a slight effect on porcine pancreatic characteristics; however, dietary protein had a greater effect on pancreatic characteristics in market-weight hogs.