Effect of body fat mass and nutritional status on 24-hour leptin profiles in ewes

This study was designed to determine the effect of feeding or fasting of fat or thin ewes on 24-h leptin profiles. Ewes were assigned, based on ultrasonic assessments of last-rib subcutaneous fat measurements, into fat (fat thickness > 1 cm; mean = 1.52 +/- 0.03 cm; range 1.14 to 2.18 cm) or thin (fat thickness < 1 cm; mean = 0.25 +/- 0.03 cm; range 0.03 to 0.84 cm) groups. Fat and thin ewes were then assigned to either fed or fasted (deprived of feed) groups consisting of five ewes per group. Thus, four groups existed and were designated as fat-fed, fat-fasted, thin-fed, and thin-fasted. Fed ewes had ad libitum access to feed throughout the study. Fasted ewes were prohibited access to feed beginning 48 h preceding the experiment. Plasma samples were collected for leptin analysis from ewes every 15 min for 24 h beginning 48 h after the initiation of feed restriction or the congruent interval in fed ewes. Data were subjected to CLUSTER pulse analysis procedures. Profiles of plasma concentrations of leptin were episodic in nature and did not differ in a diurnal manner. Fed ewes had greater mean concentrations of leptin, area under the curve, number of peaks, peak height, peak nadir, and a shorter interval between peaks than fasted ewes (P < or = 0.05). Fat ewes had greater mean concentrations of leptin, area under the curve, number of peaks, peak height, peak nadir, and a shorter interval between peaks than thin ewes (P < 0.02). There also was a tendency for a body condition x treatment interaction for number of peaks (P = 0.073) and interval between peaks (P = 0.056). These results provide evidence that plasma concentrations of leptin are episodic in nature and are influenced by nutritive state and fat thickness over the ribs, but display no circadian variation.     

Effects of glucose and amino acids on ghrelin secretion in sheep

Two experiments were conducted to elucidate the effects of post‐ruminal administration of starch and casein (Exp. 1), plasma amino acids concentrations (Exp. 2), and plasma glucose and insulin concentrations (Exp. 2) on plasma ghrelin concentrations in sheep. In Exp. 1, plasma ghrelin concentrations were determined by four infusion treatments (water, cornstarch, casein and cornstarch plus casein) in four wethers. Abomasal infusion of casein increased plasma α‐amino N (AAN) concentrations. Infusion of starch or casein alone did not affect plasma ghrelin concentrations, but starch plus casein infusion increased plasma levels of ghrelin, glucose and AAN. In Exp 2, we investigated the effects of saline or amino acids on ghrelin secretion in four wethers. Two hours after the initiation of saline or amino acid infusion into the jugular vein, glucose was also continuously infused to investigate the effects of blood glucose and insulin by hyper‐glycemic clump on plasma ghrelin concentrations. Infusion of amino acids alone raised plasma levels of ghrelin, but the higher plasma glucose and insulin concentrations had no effect on plasma ghrelin concentrations. These results suggest that high plasma levels of amino acids can stimulate ghrelin secretion, but glucose and insulin do not affect ghrelin secretion in sheep.     

Effects of dietary energy levels on plasma leptin in sheep

This experiment investigates the changes in the plasma leptin levels of sheep fed a diet of three energy levels (low, moderate and high). Four mature wethers were used for this experiment. For the first 4 weeks, the sheep were fed diets to provide 1.2 times the maintenance metabolizable energy requirements, low energy levels (LE). During the second 4 weeks, the sheep were fed diets to provide 1.5 times the maintenance metabolizable energy requirements, moderate energy levels (ME). During third 4‐week period, the sheep were fed a diet to provide 1.8 times the maintenance metabolizable energy requirements, high energy levels (HE). Body weight was determined every week. Blood samples were collected prior to the morning meal at 3 days intervals throughout the experiment, and plasma leptin, insulin and nonesterified fatty acid (NEFA) concentrations were assayed. Body weight decreased in week 1 after the start of the experiment, it continued to decrease during the LE feeding, but it gradually increased until the end of HE period. Similarly, plasma leptin concentration decreased during LE feeding, but increased during the HE feeding. Additionally, positive correlation was obtained between leptin and insulin or glucose concentrations, whereas no clear relationship with circulating NEFA was observed. In conclusion, it was suggested that plasma leptin concentrations were affected by the metabolizable energy in feed.     

Effects of exogenous estradiol and progesterone on plasma concentrations of leptin in ewes in non-breeding season

Estradiol and progesterone may play a role in controlling leptin secretion by utilizing their receptors in adipocytes and the genomic mechanisms of the leptin gene. This study was conducted to evaluate the effects of exogenous sex steroids on the blood leptin concentrations in ewes in the non-breeding season. Multiparous ewes were fed to maintenance level for their live weights. Blood samples were collected at 12-h intervals from Days -3 to -1 to determine the basal leptin levels (pre-injection period). From Day 0 to Day 5 (injection period), blood sampling continued at 12-h intervals, and the ewes were injected intramuscularly at 24-h intervals with oil, 50 mg progesterone in oil, 1 mg of estradiol in oil, or both steroids in oil. Leptin was measured using a sensitive and specific radioimmunoassay based on recombinant bovine leptin. Overall, plasma concentrations of leptin were not affected by any of the steroid treatments, and there were no differences in the value of leptin between the pre-injection and injection periods among the 4 groups. Therefore, the exogenous estrogen and progesterone used in this study do not have a strong effect on the blood leptin concentrations of ewes in the non-breeding season.     

Experimental and clinical studies on plasma leptin in obese dogs

Leptin is a protein synthesized and secreted primarily by adipocytes, and the circulating leptin concentration is elevated in obese humans and rodents. Recently, we have established a sandwich enzyme-linked immunosorbent assay for canine leptin. In the present study, plasma leptin concentrations were measured in experimentally developed obese beagles and in clinically obese dogs. When 5 male beagles were given a high-energy diet for 3 months, all of them became obese and the plasma leptin concentration significantly increased from 2.4+/-1.2 to 4.9+/-0.9 ng/ml, positively correlating with body fat content estimated by the deuterium oxide dilution method (r=0.87). The leptin concentrations of plasma samples collected from 59 dogs in veterinary practices were compared with their body condition scores (BCS). The plasma leptin concentrations of obese dogs were 9.7+/-0.7 and 12.3+/-1.5 ng/ml at BCS=4 and BCS=5, respectively, which were significantly higher than those of optimal (BCS=3) dogs (2.7+/-0.3 ng/ml). There was no significant effect of sex and breed. A weak positive correlation (r=0.37) was found between the plasma leptin concentration and age, probably due to the lesser content of visceral fat in puppies younger than 1 year old. These results indicate that plasma leptin is a good index of adiposity in dogs regardless of breed, age and sex, and may be useful for quantitative assessment of obesity in small animal practice.     

Effects of short-term feed deprivation and melatonin implants on circadian patterns of leptin in the horse

Leptin is a protein hormone produced by adipose tissue that influences hypothalamic mechanisms regulating appetite and energy balance. In species tested thus far, including horses, concentrations of leptin increase as animal fat mass increases. The variables and mechanisms that influence the secretion of leptin are not well known, nor is it known in equine species how the secretion of leptin is influenced by acute alterations in energy balance, circadian patterns, and/or reproductive competence. Our objectives were to determine in horses: 1) whether plasma concentrations of leptin are secreted in a circadian and/or a pulsatile pattern; 2) whether a 48-h period of feed restriction would alter plasma concentrations of leptin, growth hormone, or insulin; and 3) whether ovariectomy and/or a melatonin implant would affect leptin. In Exp. 1, mares exposed to ambient photoperiod of visible light (11 h, 33 min to 11 h, 38 min), received treatments consisting of a 48-h feed restriction (RES) or 48 h of alfalfa hay fed ad libitum (FED). Mares were maintained in a dry lot before sampling and were tethered to a rail during sampling. Analyses revealed that leptin was not secreted in a pulsatile manner, and that mean leptin concentrations were greater (P < 0.001) in FED vs. RES mares (17.20 +/- 0.41 vs. 7.29 +/- 0.41 ng/mL). Plasma growth hormone was pulsatile, and mean concentrations were greater in RES than FED mares (2.15 +/- 0.31 vs. 1.08 +/- 0.31 ng/mL; P = 0.05). Circadian patterns of leptin secretion were observed, but only in FED mares (15.39 +/- 0.58 ng/mL for morning vs. 19.00 +/- 0.58 ng/mL for evening; P < 0.001). In Exp. 2, mares that were ovariectomized or intact received either a s.c. melatonin implant or a sham implant. Thereafter, blood was sampled at weekly intervals at 1000 and 1700. Concentrations of leptin in samples collected at 1700 were greater (P < 0.001) than in those collected at 1000 (28.24 +/- 1.7 vs. 22.07 +/- 1.7 ng/mL). Neither ovariectomy nor chronic treatment with melatonin affected plasma concentrations of leptin or the circadian pattern of secretion. These data provide evidence that plasma leptin concentrations in the equine are sensitive to acute changes in nutritional status and vary in a circadian pattern that is sensitive to fasting but not to melatonin treatment or ovariectomy.     

Seasonal changes in the interactions among leptin, ghrelin, and orexin in sheep

The adaptation of the physiology of an animal to changing conditions of light and food availability is evident at the behavioral and hormonal levels. Melatonin, leptin, ghrelin, and orexin, which exhibit rhythmic secretion profiles under ad libitum feeding conditions, are sensitive to changes in daylength, forming a tight web of interrelationships in the regulation of energy balance. The aim of this study was to determine the effects of central injections of leptin, ghrelin, and orexin on the reciprocal interactions among these hormones and the influence of photoperiod on these responses. Twenty-four ovariectomized and estradiol-implanted ewes were used in a replicated switchback design. The ewes were assigned randomly to 1 of 6 treatment groups, and the treatments were infused into their third ventricles 3 times at 0, 1, and 2 h, with 0 h being at dusk. The treatments were as follows: 1) control, Ringer-Locke buffer; 2) leptin, 0.5 μg/kg BW; 3) ghrelin, 2.5 μg/kg BW; 4) orexin B, 0.3 μg/kg BW; 5) leptin antagonist, 50 μg/kg BW, then ghrelin, 2.5 μg/kg BW; and 6) leptin antagonist, 50 μg/kg BW, then orexin B, 0.3 μg/kg BW. Blood samples (5 mL) were collected at 15-min intervals for 6 h. The administration of leptin increased (P < 0.05) plasma concentrations of melatonin during short-day (ShD) photoperiods and decreased (P < 0.05) them during long-day (LD) photoperiods, whereas ghrelin decreased (P < 0.05) melatonin concentrations during ShD photoperiod, and orexin had no effect (P > 0.1). Leptin attenuated (P < 0.05) ghrelin concentrations relative to the concentration in controls during ShD. The plasma concentrations of orexin were reduced (P < 0.05) after leptin infusions during LD and ShD photoperiods; however, ghrelin had the opposite effect (P < 0.05) on orexin concentration. Orexin increased (P < 0.05) ghrelin concentrations during LD. Ghrelin and orexin concentrations were increased (P < 0.05) after leptin antagonist infusions. Our data provide evidence that the secretion of leptin, ghrelin, and orexin are seasonally dependent, with relationships that are subject to photoperiodic regulation, and that leptin is an important factor that regulates ghrelin and orexin releases in sheep.     

Kinetics and disposition of orally dosed sodium chlorate in sheep

Experiments were conducted in sheep to determine excretory characteristics of sodium chlorate after a single oral dose. In Exp. 1, lambs (n = 16; age = 8.1 ± 1.7 d; BW = 8.2 ± 1.1 kg; mean ± SD) were dosed orally with 0, 30, 60, or 90 mg/kg BW of sodium chlorate. Twenty-four hours after exposure chlorate residues were dose dependent (P < 0.05) in small intestinal contents, serum, and urine, but chlorate residues were not consistently detected in cecal or colonic contents. In Exp. 2, non-pregnant yearling ewes (BW = 74.8 ± 5.6 kg; mean ± SD) were orally dosed with 0, 150, 300, or 450 mg/kg BW of sodium chlorate. Across dose, chlorate residues averaged from 47 to 114, 0.6 to 4.5, and were not detectable to 0.2 μg/mL at 24, 48, and 72 h, respectively, in serum of treated animals; in feces, residues averaged 29 to 82, 0.8 to 14, and were not detectable to 1.2 μg/mL at the same respective time periods. In Exp. 3, six lactating ewes (BW = 76.3 ± 8.0 kg) were dosed orally with 450 mg/kg BW of sodium chlorate; residues were measured in serum, milk, urine and feces in periods encompassing 0 to 8, 8 to 16, 16 to 24, 24 to 32, 32 to 40, and 40 to 48 h. Chlorate residues in milk were detectable at all time periods with concentrations averaging from 287 ± 67 to 26 ± 13 μg/mL during the first and last collection periods, respectively. Urine contained the greatest concentration of chlorate at each time point and averaged 480 ± 268 μg/mL at 40 to 48 h. Depletion half-lives in serum, milk, urine, and feces were estimated to be 6.2, 27, 19, and 10 h, respectively; milk, urinary and fecal half-lives are likely overestimated due to the fact that 8-h sample pools were used in half-life estimations. In Exp. 4, three wethers (BW = 87.1 ± 5.3 kg) each were orally dosed with 14 or 42 mg/kg BW of sodium chlorate; blood samples were serially collected for 48 h, and urine samples were collected at 0 to 8, 8 to 16, 16 to 24, 24 to 36, and 36 to 48 h. Estimates of absorption and elimination half-lives based on serum chlorate concentrations were about 0.4 and 2.5 h, respectively. Urine collected during the 6 h immediately following dosing contained the greatest concentrations of chlorate residues relative to subsequent collection periods. Rapid removal of chlorate from the gastrointestinal lumen suggests that effects of chlorate on colonic and fecal gastrointestinal bacteria may occur through mechanisms other than direct luminal contact between microbe and chlorate salts.     

Relationship between plasma leptin concentrations and carcass composition in fattening mutton: a comparison with ultrasound results

Positive relationships between circulating leptin concentrations and body fat content have been established in sheep when covering a rather broad range of age and/or body weight. The usefulness of leptin measurements for predicting carcass fat has yet to be evaluated specifically in fattening lambs. We therefore measured plasma leptin concentrations in 56 male lambs half and half Merino Mutton and Blackheaded Mutton. Subcutaneous fat thickness was measured by ultrasound 1 day before the lambs were slaughtered at 35 or 45 kg live weight. Carcass composition was determined by tissue dissection. The coefficients of correlations between leptin and the different amounts in fat depots ranged from 0.40 to 0.56 within the two live weight groups, and from 0.53 to 0.64 when taking the two groups together. Carcass fat percentage was estimated by leptin concentrations with the same accuracy (R2 = 0.34) as with ultrasound fat thickness. The accuracy was higher for leptin in the 35 kg-group whereas the accuracy was higher for ultrasound fat thickness in the 45 kg-group (R2 = 0.26 vs. 0.31). A combination of leptin and ultrasound fat thickness clearly enhanced the precision of estimation in all groups. Further investigations on the influence of factors such as breed, gender, duration of feed withdrawal or photoperiod on the association between leptin and carcass composition are necessary before the suitability of plasma leptin concentration for practical application can be evaluated.     

Plasma leptin responses to lipopolysaccharide and tumor necrosis factor alpha in cows

Peripheral administration of bacterial lipopolysaccharide (LPS) and various inflammatory cytokines to rodents is known to raise plasma levels of leptin, a potent satiety factor secreted from adipocytes, implying a role of leptin in endotoxin-induced anorexia. We previously reported no effect of LPS on serum leptin levels in sheep, despite marked anorexia and fever. Our results suggest that leptin might not be involved in the endotoxin-induced anorexia in ruminants. To test this idea, in the present study, plasma leptin levels were measured during acute experimental endotoxemia in Holstein cows. Intravenous injection of LPS induced anorexia accompanied with increases in plasma levels of cortisol and insulin, all of which are known to stimulate leptin secretion in rodent and human, while it did not affect plasma leptin levels at all in cows. Similar results were also obtained after injection of recombinant bovine tumor necrosis factor alpha. These results indicate that plasma leptin levels in cows during acute endotoxemia are differentially regulated from those in rodents, and that leptin might not be involved in the endotoxin-induced anorexia in ruminants.     

Circulating estradiol suppresses luteinizing hormone pulse frequency during dietary restriction

The influence of dietary restriction on the negative feedback potency of 17-beta-estradiol (E2) was evaluated in both castrated male (wethers) and female sheep (OVX ewes) during the breeding season. In study 1, OVX ewes received maintenance or restricted dietary energy for 7 weeks or maintenance energy for 6 weeks prior to a 5 day fast (n=12ewes/feeding group). Estradiol (0.31microg E2/50kg/h) or vehicle (10% EtOH-saline) was continuously infused into half the animals in each dietary treatment for the final 54h of the study. The dynamic pattern of LH secretion was assessed during the final 6h of infusion. Estradiol inhibited luteinizing hormone (LH) pulse amplitude independent of nutrition (P=0.02); fasting increased mean LH, LH peak height, and LH nadir in the absence of E2 (P=0.004, P=0.02, and P=0.02, respectively); while E2 inhibited pulse frequency (P=0.02) and increased peak width (P=0.04) in restricted ewes. Interestingly, despite uniform E2 delivery, serum concentrations of E2 differed with feeding status. Therefore, 12 wethers were infused with 0.31microg E2/50kg/h (6 fed, 6 fasted) and six wethers received 0.19microg E2/50kg/h (fasted) to establish similar serum concentrations of E2 in fed (0.31microg/50kg/h) and fasted (0.19microg/50kg/h) wethers. When fed and fasted wethers had uniform serum concentrations of E2 LH pulse frequency was suppressed (P<0.05) in fasted relative to fed animals, supporting the postulate that energy restriction enhances the E2 negative feedback potency. Collectively, these studies demonstrate that nutrition affects E2 feedback potency and clearance.     

Nutrient digestibility in sheep fed diets containing Roundup Ready or conventional fodder beet, sugar beet, and beet pulp

The objective of this digestibility assessment was to determine whether there are significant differences in the digestibility of Roundup Ready (glyphosate-tolerant) and conventional sugar beet, fodder beet, and beet pulp produced from sugar beet varieties when fed to sheep (seven wethers per treatment group). Three experiments were conducted in this assessment. Experiment 1 (35 wethers) compared one glyphosate-tolerant fodder beet variety with four conventional varieties, Exp. 2 (42 wethers) compared one glyphosate-tolerant sugar beet variety with five conventional varieties, and Exp. 3 (42 wethers) compared beet pulp derived from glyphosate-tolerant sugar beet with beet pulp from five European locations. The experimental phase consisted of a 2-wk preliminary period followed by a 1-wk collection period for Exp. 1 and 2, and a 1-wk preliminary period followed by a 1-wk digestibility collection period for Exp. 3. Diets were comprised of grass hay at 30, 30, and 20% of DM for Exp. 1, 2, and 3, respectively, with the balance being beet components. Urea and sodium sulfate were supplemented (8 and 2.9 g, respectively, for Exp. 1 and 2; and 6 g and 2.16 g, respectively, for Exp. 3) to supply sufficient dietary N and S. Each diet was fed to sheep (96 +/- 0.9 kg) in the three experiments to at or near maintenance energy levels. Treatment differences were considered significant at P < 0.05. Apparent digestibilities of DM, OM, CP, NDF, ADF, and DE for glyphosate-tolerant fodder and sugar beets did not differ from those for commercial fodder and sugar beets in Exp. 1 and 2. There were differences (P < 0.05) in DM, OM, CP, NDF, ADF, and DE digestibilities influenced by the different varieties of beet pulp in Exp. 3, but these were not unique to just the Roundup Ready sugar beet variety. Digestibilities and feeding values of Roundup Ready fodder beet, sugar beet, and beet pulp produced from Roundup Ready sugar beet varieties were not influenced by the introduction of the Roundup Ready trait compared with conventional varieties.     

Effect of feeding fat or intrajugular infusion of glucagon-like peptide-1 and cholecystokinin on dry matter intake, digestibility, and digesta rate of passage in growing wethers

A cause and effect relationship between glucagon-like peptide 1 (7, 36) amide (GLP-1) and cholecystokinin (CCK) and DMI regulation has not been established in ruminants. Three randomized complete block experiments were conducted to determine the effect of feeding fat or infusing GLP-1 or CCK intravenously on DMI, nutrient digestibility, and Cr rate of passage (using Cr(2)O(3) as a marker) in wethers. A total of 18 Targhee × Hampshire wethers (36.5 ± 2.5 kg of BW) were used, and each experiment consisted of four 21-d periods (14 d for adaptation and 7 d for infusion and sampling). Wethers allotted to the control treatments served as the controls for all 3 experiments; experiments were performed simultaneously. The basal diet was 60% concentrate and 40% forage. In Exp. 1, treatments were the control (0% added fat) and addition of 4 or 6% Ca salts of palm oil fatty acids (DM basis). Treatments in Exp. 2 and 3 were the control and 3 jugular vein infusion dosages of GLP-1 (0.052, 0.103, or 0.155 μg?kg of BW(-1)?d(-1)) or CCK (0.069, 0.138, or 0.207 μg?kg of BW(-1)?d(-1)), respectively. Increases in plasma GLP-1 and CCK concentrations during hormone infusions were comparable with increases observed when increasing amounts of fat were fed. Feeding fat and infusion of GLP-1 tended (linear, P = 0.12; quadratic, P = 0.13) to decrease DMI. Infusion of CCK did not affect (P > 0.21) DMI. Retention time of Cr in the total gastrointestinal tract decreased (linear, P < 0.01) when fat was fed, but was not affected by GLP-1 or CCK infusion. In conclusion, jugular vein infusion produced similar plasma CCK and GLP-1 concentrations as observed when fat was fed. The effects of feeding fat on DMI may be partially regulated by plasma concentration of GLP-1, but are not likely due solely to changes in a single hormone concentration.     

Dietary restriction reduces the rate of estradiol clearance in sheep (Ovis aries)

Three experiments were designed to test the effect of dietary restriction on clearance of 17beta-estradiol (E(2)) in sheep. A preliminary experiment examined the effect of a 4-d fast on the rate of E(2) clearance in wethers. The second experiment tested the hypothesis that either long-term restriction (7 wk) or a 5-d fast would increase steroid-binding capacity of serum by increasing the concentration of sex hormone-binding globulin (SHBG) in the blood of ovariectomized ewes. In Exp. 3, we hypothesized that nutrition-dependent regulation of E(2) clearance by the liver would result in divergence in biliary extraction of E(2) in fed and fasted wethers receiving comparable levels of exogenous E(2). A marked difference in E(2) clearance between fed and fasted wethers was noted in the preliminary study. Relative to ad libitumfed wethers, a 4-d fast decreased E(2) clearance by 52%. Serum concentrations of SHBG were increased in long-term energy-restricted and fasted ewes, relative to the concentration in maintenancefed ewes (P = 0.015). Furthermore, a 5-d fast nearly doubled serum steroid-binding capacity in wethers. The E(2) concentration in bile was 2 times greater in fasted than in fed wethers. This fasting-dependent increase in biliary E(2) may be reflective of the increased serum E(2) in fasted animals, because each 1 pg/mL increase in serum E(2) increased bile E(2) by 0.86 +/- 0.12 pg/mL, independent of nutrition (P = 0.002). Our results demonstrate that the rate of clearance of E(2) is decreased during nutritional restriction. Additionally, these data indicate that altered SHBG expression, enterohepatic recirculation, or both are involved in the decreased E(2) clearance during dietary restriction.     

Evaluation of the Captec chrome controlled-release device for the estimation of fecal output by grazing sheep.

Three grazing experiments were conducted to compare actual fecal output (FO) by sheep, determined with fecal collection bags, to fecal output predicted using Captec chromic oxide (Cr2O3) controlled-release capsules. In Exp. 1, 14 crossbred wethers dosed with controlled-release Cr2O3 capsules rotationally grazed four paddocks of alfalfa (Medicago sativa L.) for 18 d. Total fecal collection and grab samples were taken daily from each animal; feces were dried and assayed for Cr. From d 5 to 12 after dosing, mean actual FO (429 +/- 9.6 g of DM/d) differed (P less than .001) from FO predicted by the capsule (463 +/- 12.8 g of DM/d). The correlation between actual and predicted FO was r = .59. When data were averaged by day, the correlation increased to .82. In Exp. 2, Tifton 44 bermudagrass (Cynodon dactylon [L.] Pers.) was strip-grazed for 30 d by the same 14 wethers used in Exp. 1. Mean actual FO (369 +/- 5.1 g of DM/d) differed (P less than .0001) from predicted FO (415 +/- 9.3 g of DM/d), with a correlation of .60. In Exp. 3, 72 crossbred lambs were blocked by weight (light and heavy) and assigned to six groups of eight wethers (four with fecal bags) and four ewes, allotted randomly to three stocking rates (74, 99, and 148 sheep/ha). Sampling occurred from d 6 to 10 after dosing (Period 1) and d 20 to 24 after dosing (Period 2).(ABSTRACT TRUNCATED AT 250 WORDS)     

AMP-activated protein kinase and adipogenesis in sheep fetal skeletal muscle and 3T3-L1 cells

Marbling, or i.m. fat, is an important factor determining beef quality. Both adipogenesis and hypertrophy of existing adipocytes contribute to enhanced marbling. We hypothesized that the fetal stage is important for the formation of i.m. adipocytes and that AMP-activated protein kinase (AMPK) has a key role in adipogenesis during this stage. The objective of this study was to assess the role of AMPK in adipogenesis in fetal sheep muscle and 3T3-L1 cells. Nonpregnant ewes were randomly assigned to a control (Con, 100% of NRC recommendations, n = 7) or overfed (OF, 150% of NRC, n = 7) diet from 60 d before to 75 d after conception, when the ewes were killed. The fetal LM was collected at necropsy for biochemical analyses. The activity of AMPK was less in the fetal muscle of OF sheep. The expression of peroxisome proliferator-activated receptor (PPAR)gamma, a marker of adipogenesis, was greater in OF fetal muscle compared with Con fetal muscle. To further show the role of AMPK in adipogenesis, we used 3T3-L1 cells. The 3T3-L1 cells were incubated in a standard adipogenic medium for 24 h and 10 d. Activation of AMPK by 5-aminoimidazole-4-car-boxamide-1-beta-d-ribonucleoside dramatically inhibited the expression of PPARgamma and reduced the presence of adipocytes after 10 d of differentiation. Inhibition of AMPK by compound C enhanced the expression of PPARgamma. In conclusion, these data show that AMPK activity is inversely related to adipogenesis in fetal sheep muscle and 3T3-L1 cells.     

Leptin in ruminants. Gene expression in adipose tissue and mammary gland, and regulation of plasma concentration

This paper reviews data on leptin gene expression in adipose tissue (AT) and mammary gland of adult ruminants, as well as on plasma leptin variations, according to genetic, physiological, nutritional and environmental factors. AT leptin mRNA level was higher in sheep and goat subcutaneous than visceral tissues, and the opposite was observed in cattle; it was higher in fat than in lean selection line in sheep; it was decreased by undernutrition and increased by refeeding in cattle and sheep, and not changed by adding soybeans to the diet of lactating goats; it was increased by injection of NPY to sheep, and by GH treatment of growing sheep and cattle. Insulin and glucocorticoids in vitro increased AT leptin mRNA in cattle, and leptin production in sheep. Long daylength increased AT lipogenic activities and leptin mRNA, as well as plasma leptin in sheep. Mammary tissue leptin mRNA level was high during early pregnancy and was lower but still expressed during late pregnancy and lactation in sheep. Leptin was present in sheep mammary adipocytes, epithelial and myoepithelial cells during early pregnancy, late pregnancy and lactation, respectively. Plasma leptin in cattle and sheep was first studied thanks to a commercial “multi-species” kit. It was positively related to body fatness and energy balance or feeding level, and decreased by β-agonist injection. The recent development of specific RIA for ruminant leptin enabled more quantitative study of changes in plasma leptin concentration, which were explained for 35–50% by body fatness and for 15–20% by feeding level. The response of plasma leptin to meal intake was related positively to glycemia, and negatively to plasma 3-hydroxybutyrate. The putative physiological roles of changes in leptin gene expression are discussed in relation with published data on leptin receptors in several body tissues, and on in vivo or in vitro effects of leptin treatment.     

Palatability of wethers fed an 80% barley diet processed at different ages and of yearling wethers grazed on native range

Seasonal availability of lamb in the Western United States contributes to a large fluctuation in lamb supply and value. However, alternatives to fall marketing may not be practical unless palatability traits are acceptable. A 3-yr study was conducted to investigate 1) the effects of slaughter age (7 to 8; 10 to 11; or 14 to 15 mo) on carcass and palatability characteristics of wethers fed an 80% barley diet (Exp. 1); and 2) the effects of finishing on range or on an 80% barley diet on carcass and palatability traits of 14- to 15-mo-old wethers (Exp. 2). In Exp. 1, no differences (P = .27) were detected in flavor intensity or longissimus muscle area among slaughter age groups, but fat depth was greater (P < .05) for 7- to 8-mo-old wethers than for 10- to 11- or 14- to 15-mo-old wethers. Year x slaughter age interactions were detected (P < .10) for hot carcass weight, Warner-Bratzler shear value, body wall thickness, and percentage kidney fat. Hot carcass weight was greater (P < .05) for 14- to 15-mo-old wethers than for both groups of younger wethers in yr 1, did not differ (P = .53) among slaughter ages in yr 2, and was greater (P < .05) for 10- to 11- than for 14- to 15-mo-old wethers in yr 3. Warner-Bratzler shear values did not differ (P > .10) among slaughter ages in yr 1 and 3, but shear values for 14- to 15-mo-old wethers were greater (P < .05) than for both younger slaughter age groups in yr 2. Percentage kidney fat was lower (P < .05) for 14- to 15- than for 7- to 8-mo-old wethers in all years. In Exp. 2, flavor intensity of the meat did not differ (P = .35) between finishing systems, but longissimus muscle area was greater (P = .02) for range-finished wethers than for wethers fed an 80% barley diet. Year x finishing treatment interactions were detected (P < .10) for shear values, body wall thickness, percentage kidney fat, and fat depth. Shear values were greater (P = .10) for range-finished wethers than for wethers fed an 80% barley diet in yr 1, but did not differ (P > .55) in yr 2 and 3. Body wall and fat measurements were greater (P < .10) for wethers fed an 80% barley diet than for range-finished wethers in all years except yr 3, when fat depth did not differ (P = .47). Overall, slaughtering wethers fed an 80% barley diet or range-finished wethers at older ages produced acceptable carcasses with desirable meat palatability traits.     

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.