N-methyl-d,l-aspartate modulation of pituitary hormone secretion in the pig: Role of opioid peptides

Sixteen ovariectomized (OVX) mature gilts, averaging 139.6 ± 3.1 kg body weight (BW) were assigned randomly to receive either progesterone (P, 0.85 mg/kg BW, n=8) or corn oil vehicle (OIL, n=8) injections im twice daily for 10 d. On the day of experiment, all gilts received either the EAA agonist, N-methyl-d,l-aspartate (NMA; 10 mg/kg BW, iv) alone or NMA plus the EOP antagonist, naloxone (NAL, 1 mg/kg BW, iv), resulting in the following groups of 4 gilts each: OIL-NMA, OIL-NMA-NAL, P-NMA and P-NMA-NAL. Blood samples were collected via jugular cannula every 15 min for 6 hr. All pigs received NMA 5 min following pretreatment with either 0.9% saline or NAL 2 hr after blood collection began and a GnRH challenge 3 hr after NMA. Administration of NMA suppressed (P<0.03) LH secretion in OIL-NMA gilts and treatment with NAL failed to reverse the suppressive effect of NMA on LH secretion in OIL-NMA-NAL gilts. Similar to OIL-NMA gilts, NMA decreased (P<0.03) mean serum LH concentrations in P-NMA gilts. However, in P-NMA-NAL gilts, serum LH concentrations were not changed following treatment. All gilts responded to GnRH with increased (P<0.01) LH secretion. Additionally, administration of NMA increased (P<0.01) growth hormone (GH) and prolactin (PRL) secretion in both OIL-NMA and P-NMA gilts, but this increase in GH and PRL secretion was attenuated (P<0.01) by pretreatment with NAL in OIL-NMA-NAL and P-NMA-NAL gilts. Serum cortisol concentrations increased (P<0.01) in all gilts and the magnitude of the cortisol response was not different among groups. In summary, results of the present study confirmed previous findings that NMA suppresses LH secretion in both oil- and P-treated OVX gilts, but we failed to provide definitive evidence that EOP are involved in the NMA-induced suppression of LH secretion. However, NMA may, in part, activate the EOP system which in turn increased GH and PRL secretion in the gilt.     

Nutritionally-induced anestrus in gilts: metabolic and endocrine changes associated with cessation and resumption of estrous cycles

Two experiments determined how feed restriction and realimentation altered metabolism and ovarian function in gilts. In Exp. 1, cyclic (INTACT-R, n=6) and ovariectomized (OVEX-R, n=6) gilts were fed restricted diets (.23 kg feed.d-1) or ovariectomized (OVEX-C, n=6) gilts were fed control diets (1.81 kg.d-1). Estrous cycles stopped after 46 +/- 9 d of feed restriction. Average weight (WT), backfat thickness (BF) and concentrations of insulin (INS) were lower and free fatty acids (FFA) were greater in OVEX-R than in OVEX-C gilts. Frequency of luteinizing hormone (LH) release (peaks.6 h-1) was reduced by feed restriction (.2 +/- .2, 1.8 +/- 1.0 and 5.8 +/- .2 in INTACT-R, OVEX-R and OVEX-C gilts, respectively). Patterns of secretion of LH and follicle stimulating hormone (FSH) after gonadotropin releasing hormone (GnRH) or estradiol benzoate were not altered by feed restriction. Feed intake was then increased in INTACT-R and OVEX-R gilts beginning on d 80 and 82, respectively. Resumption of estrous cycles in INTACT-R gilts occurred on d 116.0 +/- 4.0 and was preceded by a significant increase in WT, but not BF, and a linear increase in concentration and frequency of release of LH. Increasing feed intake in OVEX-R gilts increased WT and frequency of LH release, while FFA decreased and INS increased to concentrations not different from those of OVEX-C gilts. The hypothesis that nutritionally-induced anestrus resulted from decreased activity of the hypothalamic pulse-generator was evaluated in Exp. 2 by providing 144 hourly pulses (iv) of saline (n=3), GnRH (n=3) or LH (n=4) to nutritionally-anestrous gilts.(ABSTRACT TRUNCATED AT 250 WORDS)     

N-methyl-d,l-aspartate stimulates growth hormone and prolactin but inhibits luteinizing hormone secretion in the pig.

The effects of n-methyl-d,l-aspartate (NMA), a neuroexcitatory amino acid agonist, on luteinizing hormone (LH), prolactin (PRL) and growth hormone (GH) secretion in gilts treated with ovarian steroids was studied. Mature gilts which had displayed one or more estrous cycles of 18 to 22 d were ovariectomized and assigned to one of three treatments administered i.m.: corn oil vehicle (V; n = 6); 10 micrograms estradiol-17 b/kg BW given 33 hr before NMA (E; n = 6); .85 mg progesterone/kg BW given twice daily for 6 d prior to NMA (P4; n = 6). Blood was collected via jugular cannulae every 15 min for 6 hr. Pigs received 10 mg NMA/kg BW i.v. 2 hr after blood collection began and a combined synthetic [Ala15]-h GH releasing factor (1-29)-NH2 (GRF; 1 micrograms/kg BW) and gonadotropin releasing hormone (GnRH; .2 micrograms/kg BW) challenge given i.v. 3 hr after NMA. NMA did not alter LH secretion in E gilts. However, NMA decreased (P < .02) serum LH concentrations in V and P4 gilts. Serum LH concentrations increased (P < .01) after GnRH in all gilts. NMA did not alter PRL secretion in P4 pigs, but increased (P < .01) serum PRL concentrations in V and E animals. Treatment with NMA increased (P < .01) GH secretion in all animals while the GRF challenge increased (P < .01) serum GH concentrations in all animals except in V treated pigs. NMA increased (P < .05) cortisol secretion in all treatment groups. These results indicate that NMA inhibits LH secretion and is a secretagogue of PRL, GH and cortisol secretion with ovarian steroids modulating the LH and PRL response to NMA.     

Serum leptin concentrations, luteinizing hormone and growth hormone secretion during feed and metabolic fuel restriction in the prepuberal gilt

Two experiments were conducted to determine 1) the effect of acute feed deprivation on leptin secretion and 2) if the effect of metabolic fuel restriction on LH and GH secretion is associated with changes in serum leptin concentrations. Experiment (EXP) I, seven crossbred prepuberal gilts, 66 +/- 1 kg body weight (BW) and 130 d of age were used. All pigs were fed ad libitum. On the day of the EXP, feed was removed from four of the pigs at 0800 (time = 0) and pigs remained without feed for 28 hr. Blood samples were collected every 10 min from zero to 4 hr = Period (P) 1, 12 to 16 hr = P 2, and 24 to 28 hr = P 3 after feed removal. At hr 28 fasted animals were presented with feed and blood samples collected for an additional 2 hr = P 4. EXP II, gilts, averaging 140 d of age (n = 15) and which had been ovariectomized, were individually penned in an environmentally controlled building and exposed to a constant ambient temperature of 22 C and 12:12 hr light: dark photoperiod. Pigs were fed daily at 0700 hr. Gilts were randomly assigned to the following treatments: saline (S, n = 7), 100 (n = 4), or 300 (n = 4) mg/kg BW of 2-deoxy-D-glucose (2DG), a competitive inhibitor of glycolysis, in saline iv. Blood samples were collected every 15 min for 2 hr before and 5 hr after treatment. Blood samples from EXP I and II were assayed for LH, GH and leptin by RIA. Selected samples were quantified for glucose, insulin and free fatty acids (FFA). In EXP I, fasting reduced (P < 0.04) leptin pulse frequency by P 3. Plasma glucose concentrations were reduced (P < 0.02) throughout the fast compared to fed animals, where as serum insulin concentrations did not decrease (P < 0.02) until P 3. Serum FFA concentrations increased (P < 0.02) by P 2 and remained elevated. Subcutaneous back fat thickness was similar among pigs. Serum IGF-I concentration decreased (P < 0.01) by P 2 in fasted animals compared to fed animals and remained lower through periods 3 and 4. Serum LH and GH concentrations were not effected by fast. Realimentation resulted in a marked increase in serum glucose (P < 0.02), insulin (P < 0.02), serum GH (P < 0.01) concentrations and leptin pulse frequency (P < 0.01). EXP II treatment did not alter serum insulin levels but increased (P < 0.01) plasma glucose concentrations in the 300 mg 2DG group. Serum leptin concentrations were 4.0 +/- 0.1, 2.8 +/- 0.2, and 4.9 +/- 0.2 ng/ml for S, 100 and 300 mg 2DG pigs respectively, prior to treatment and remained unchanged following treatment. Serum IGF-I concentrations were not effected by treatment. The 300 mg dose of 2DG increased (P < 0.0001) mean GH concentrations (2.0 +/- 0.2 ng/ml) compared to S (0.8 +/- 0.2 ng/ml) and 100 mg 2DG (0.7 +/- 0.2 ng/ml). Frequency and amplitude of GH pulses were unaffected. However, number of LH pulses/5 hr were decreased (P < 0.01) by the 300 mg dose of 2DG (1.8 +/- 0.5) compared to S (4.0 +/- 0.4) and the 100 mg dose of 2DG (4.5 +/- 0.5). Mean serum LH concentrations and amplitude of LH pulses were unaffected. These results suggest that acute effects of energy deprivation on LH and GH secretion are independent of changes in serum leptin concentrations.     

Recombinant Porcine Leptin Reduces Feed Intake and Stimulates Growth Hormone Secretion in Swine

Two experiments (EXP) were conducted to test the hypothesis that porcine leptin affects GH, insulin-like growth factor-I (IGF-I), insulin, thyroxine (T₄) secretion, and feed intake. In EXP I, prepuberal gilts received intracerebroventricular (ICV) leptin injections. Blood was collected every 15 min for 4 hr before and 3 hr after ICV injections of 0.9% saline (S; n = 3), 10 μg (n = 4), 50 μg (n = 4), or 100 μg (n = 4) of leptin in S. Pigs were fed each day at 0800 and 1700 hr over a 2-wk period before the EXP. On the day of the EXP, pigs were fed at 0800 hr and blood sampling started at 0900 h. After the last sample was collected, feeders were placed in all pens. Feed intake was monitored at 4, 20, and 44 hr after feed presentation. In EXP II, pituitary cells from prepuberal gilts were studied in primary culture to determine if leptin affects GH secretion at the level of the pituitary. On Day 4 of culture, 10⁵ cells/well were challenged with 10⁻¹², 10⁻¹⁰, 10⁻⁸, or 10⁻⁶ M [Ala¹⁵]-h growth hormone-releasing factor-(1-29)NH₂ (GRF), 10⁻¹⁴, 10⁻¹³, 10⁻¹², 10⁻¹¹, 10⁻¹⁰, 10⁻⁹, 10⁻⁸, 10⁻⁷, or 10⁻⁶ M leptin individually or in combinations with 10⁻⁸ and 10⁻⁶ M GRF. Secreted GH was measured at 4 hr after treatment. In EXP I, before injection, serum GH concentrations were similar. Serum GH concentrations increased (P < 0.01) after injection of 10 μg (21 ± 1 ng/ml), 50 μg (9 ± 1 ng/ml), and 100 μg (13 ± 1 ng/ml) of leptin compared with S (1 ± 2 ng/ml) treated pigs. The GH response to leptin was greater (P < 0.001) in 10 μg than 50 or 100 μg leptin-treated pigs. By 20 hr the 10, 50, and 100 μg doses of leptin reduced feed intake by 53% (P < 0.08), 76%, and 90% (P < 0.05), respectively, compared with S pigs. Serum IGF-I, insulin, T₄, glucose, and free fatty acids were unaffected by leptin treatment. In EXP II, relative to control (31 ± 2 ng/well), 10⁻¹⁰, 10⁻⁸, and 10⁻⁶ M GRF increased (P < 0.01) GH secretion by 131%, 156%, and 170%, respectively. Only 10⁻⁶ M and 10⁻⁷ M leptin increased (P < 0.01) GH secretion. Addition of 10⁻¹¹ and 10⁻⁹ M leptin in combination with 10⁻⁶ M GRF or 10⁻¹¹ M leptin in combination with 10⁻⁸ M GRF-suppressed (P < 0.05) GH secretion. These results indicate that leptin modulates GH secretion and, as shown in other species, leptin suppressed feed intake in the pig.     

Opioid modulation of gonadotropin releasing hormone release from the hypothalamic preoptic area in the pig

Two experiments (Exp) were conducted to examine in vitro the release of gonadotropin releasing hormone (GnRH) from the hypothalamus after treatment with naloxone (NAL) or morphine (MOR). In Exp 1, hypothalamic-preoptic area (HYP-POA) collected from 3 market weight gilts at sacrifice and sagitally halved were perifused for 90 min prior to a 10 min pulse of morphine (MOR; 4.5 × 10⁻⁶ M) followed by NAL (3.1 × 10⁻⁵ M) during the last 5 min of MOR (MOR + NAL; N=3). The other half of the explants (n=3) were exposed to NAL for 5 min. Fragments were exposed to KCl (60 mM) at 175 min to assess residual GnRH releasability. In Exp 2, nine gilts were ovariectomized and received either oil vehicle im (V; n=3); 10 μg estradiol-17β/kg BW im 42 hr before sacrifice (E; n=3); .85 mg progesterone/kg BW im twice daily for 6 d prior to sacrifice (P₄; n=3). Blood was collected to assess pituitary sensitivity to GnRH (.2 μg/kg BW) on the day prior to sacrifice. On the day of sacrifice HYP-POA explants were collected and treated as described in Exp 1 except tissue received only NAL. In Exp 1, NAL increased (P<.05) GnRH release. This response to NAL was attenuated (P<.05) by coadministration of MOR. Cumulative GnRH release after NAL was greater (P<.05) than after MOR + NAL. All tissues responded similarly to KCl with an increase (P<.05) in GnRH release. In Exp 2, pretreatment luteinizing hormone (LH) concentrations were lower (P<.05) in E gilts compared to V and P₄ animals with P₄ being lower (P<.05) than V gilts. LH response to GnRH was lower (P<.05) in E pigs than in V and P₄ animals, while the responses was similar between V and P₄ gilts. NAL increased GnRH release in all explants, whereas, KCl increased GnRH release in 6 of 9 explants. These results indicate that endogenous opioid peptides may modulate in vitro GnRH release from the hypothalamus in the gilt.     

Luteinizing hormone release after administration of the gonadotropin-releasing hormone agonist Fertilan (goserelin) for synchronization of ovulation in pigs

The generic GnRH agonist, Fertilan (goserelin), was tested for the ability to induce an LH surge and ovulation in estrus-synchronized gilts. Three experiments were performed to 1) examine the effect of various doses of Fertilan on secretion of LH in barrows, to select doses to investigate in gilts (Exp. 1); 2) determine doses of Fertilan that would induce a preovulatory-like rise of LH in gilts (Exp. 2); and 3) determine the time of ovulation after Fertilan treatment (Exp. 3). In Exp. 1, 10 barrows were injected on d 1, 4, 7, 10, and 13 with 10, 20, or 40 microg of Fertilan; 50 microg of Gonavet (depherelin; GnRH control) or saline (negative control); and sequential blood samples were collected for 480 min. There was a dose-dependent stimulation (P < 0.05) of LH release. Maximal plasma concentrations of LH (LH(MAX)) were 2.1 +/- 0.2, 4.1 +/- 0.3, 2.6 +/- 0.4, and 3.4 +/- 0.3 ng/mL after 10, 20, and 40 microg of Fertilan and 50 microg of Gonavet, respectively, and duration of release was 78 +/- 9, 177 +/- 12, 138 +/- 7, and 180 +/- 11 min, respectively. Fertilan doses of 10 and 20 microg were deemed to be the most suitable for testing in gilts. In Exp. 2, 12 gilts received (after estrus synchronization with Regumate and eCG) injections of 10 or 20 microg of Fertilan or 50 microg of Gonavet 80 h after eCG to stimulate a preovulatory-like LH surge and ovulation. An LH surge was induced in 3 of the 4 gilts in both of the Fertilan groups and in all of the Gonavet-treated gilts. Characteristics of induced release of LH did not differ among groups: LH(MAX), 5.0 +/- 0.9 vs. 4.6 +/- 1.8 vs. 6.6 +/- 1.1 ng/mL; duration, 11.7 +/- 2.0 vs. 12.3 +/- 2.2 vs. 14.3 +/- 0.5 h; interval from GnRH injection to LH(MAX), 4.0 +/- 2.0 vs. 6.7 +/- 1.3 vs. 5.8 +/- 1.6 h. In Exp. 3, estrus-synchronized gilts were injected with 20 microg of Fertilan (n = 8) or 50 microg of Gonavet (n = 4), and the time of ovulation was determined by repeated endoscopic examination. Time of ovulation ranged from 34 to 42 h postGnRH; however, ovulation occurred earlier in the Gonavet compared with the other groups (P < 0.05). Results of these experiments indicate that 1) barrows are an appropriate model for determining GnRH doses that can be effective in inducing a preovulatory-like LH surge in females; 2) the generic GnRH agonist Fertilan, at doses of 10 to 20 microg, can stimulate an LH surge in gilts, with subsequent ovulation; and 3) Fertilan at doses of 10 and 20 microg should be examined further for use in fixed-time insemination protocols.     

The influence of exogenous pituitary growth hormone on the ovulatory response to PMSG and hCG and the LH response to estradiol benzoate in prepubertal gilts

Two experiments were performed to examine the influence of exogenous growth hormone on the reproductive axis in gilts. Experiment one employed 26 Yorkshire × Landrace prepubertal gilts, which were selected at 150 d and 86.5 ± 1.5 kg bodyweight (BW) and assigned equally to two treatments. Gilts received injections of either porcine growth hormone at 90 μg/kg BW, or vehicle buffer, from 150 to 159 d. At 154 d gilts received 500 IU PMSG, followed 96 hr later by 250 IU hCG. Gilts were slaughtered at 163 days and their ovaries recovered to determine ovulatory status. In each treatment, gilts failed to show any ovarian response to PMSG/hCG. All remaining control gilts ovulated and their ovaries appeared morphologically normal. In gilts receiving exogenous growth hormone, fewer ovaries (4/11, P<.01) appeared morphologically normal. The ovaries of all other growth hormone injected gilts had very large (12–25 mm) non-luteinized follicles. In experiment two, 20 prepubertal Yorkshire × Landrace gilts were selected at 138 days and 85 kg BW. These gilts received injections of growth hormone at 90 μg/kg BW (n=9) or vehicle (n=11) from 138 to 147 days. At 143 days, all gilts were given an injection of estradiol benzoate (EB) at 15 μg/kg BW. Blood samples were taken at the time of EB injection, at 24 and 36 hr and then at 6 hr intervals until 78 hr. All samples were assayed for serum LH concentrations. The EB induced LH peak height was lower (P<.04) in gilts receiving exogenous growth hormone than in controls. The results presented indicate that the daily injection of growth hormone at 90 μg/kg BW reduced the estradiol-induced release of LH in addition to reducing the number of corpora lutea in gonadotrophin stimulated gilts.     

Effects of cimaterol administration on plasma concentrations of various hormones and metabolites in friesian steers

The aim of the experiment was to determine the acute and chronic effects of the β-agonist, cimaterol, on plasma hormone and metabolite concentrations in steers. Twelve Friesian steers (liveweight = 488 ± 3 kg) were randomly assigned to receive either 0 (control; n=6) or .09 mg cimaterol/kg body weight/day (treated; n=6). Steers were fed grass silage ad libitum. Cimaterol, dissolved in 140 ml of acidified distilled water (pH 4.2), was administered orally at 1400 hr each d. After 13 d of treatment with cimaterol or vehicle (days 1 to 13), all animals were treated with vehicle for a further 7 d (days 14 to 20). On days 1, 13 and 20, blood samples were collected at 20 min-intervals for 4 hr before and 8 hr after cimaterol or vehicle dosing. All samples were assayed for growth hormone (GH) and insulin, while samples taken at −4, −2, 0, +2, +4, +6 and +8 hr relative to dosing were assayed for thyroxine (T₄), triiodothyronine (T₃), cortisol, urea, glucose and non-esterified fatty acids (NEFA). Samples taken at −3 and +3 hr relative to dosing were assayed for IGF-I only. On day 1, cimaterol acutely reduced (P<.05) GH and urea concentrations (7.6 vs 2.9 ± 1.4 ng/ml; and 6.0 vs 4.9 ± 0.45 mmol/l, respectively; mean control vs mean treated ± pooled standard error of difference), and increased (P<.05) NEFA, glucose and insulin concentrations (160 vs 276 ± 22 μmol/l, 4.1 vs 6.2 ± 0.15 mmol/l and 29.9 vs 179.7 ± 13.9 μU/ml, respectively). Plasma IGF-I, T₃, T₄ and cortisol concentrations were not altered by treatment. On day 13, cimaterol increased (P<.05) GH and NEFA concentrations (7.7 vs 14.5 ± 1.4 ng/ml and 202 vs 310 ± 22 mEq/l, respectively) and reduced (P<.05) plasma IGF-I concentrations (1296 vs 776 ± 227 ng/ml). Seven-d withdrawal of cimaterol (day 20) returned hormone and metabolite concentrations to control values. It is concluded that : 1) cimaterol acutely increased insulin, glucose and NEFA and decreased GH and urea concentrations, 2) cimaterol chronically increased GH and NEFA and decreased IGF-I concentrations, and 3) there was no residual effect of cimaterol following a 7-d withdrawal period.     

The effect of infusion of urea into the vena cava on feed intake of finishing gilts

Three experiments were conducted to evaluate the relationship between feed intake and plasma urea concentration. In Exp. 1, six gilts (BW 53 kg) with catheters in their venae cavae were used in a 5x5+1 Latin square design to determine the amount of infused urea needed to mimic the plasma urea concentration of pigs fed a 25% CP diet. Five gilts were fed a 16% CP corn-soybean meal diet and were infused continuously with either saline or one of four doses of urea (6, 12, 18, and 24 g/d) during each of five periods (12 h/period). Between periods, infusions were stopped for 36 h. The sixth pig was fed a 25% CP diet and infused with saline during each of the experimental periods. Venous blood samples were obtained at 1-h intervals starting 1 h before infusion. As expected, plasma urea concentration increased with increasing amount of urea infused. A daily infusion of 24 g of urea resulted in a plasma urea concentration similar to that of the pig fed the 25% CP diet with saline infusion. In Exp. 2, 12 gilts (BW 60 kg) were used in a crossover design. Pigs received a 16% CP diet and a different treatment (saline or 24 or 30 g/d of urea) in each of three infusion periods. Each infusion period lasted 2 wk. Infusions were stopped for 2 d between periods. Blood samples were obtained before infusion and daily after infusions started. Feeders were weighed daily to determine ADFI. Experiment 3 was similar to Exp. 2, except that only two treatments (saline and 30 g/d of urea) were used. Data from Exp. 2 and 3 were combined for statistical analysis. Plasma urea concentration increased linearly (P<.001) with increasing amount of urea infused. Overall, there was a trend (P<.10) for urea infusion to decrease ADFI, and pigs infused with 30 g/d consumed less (P<.05) feed than pigs infused with saline. Therefore, plasma urea concentration may play a role in regulating feed intake in gilts consuming excess protein.     

Effects of restricted feed intake on finishing pigs weighing between 68 and 114 kilograms fed twice or 6 times daily

In a previous study with limit-fed gestating gilts, we observed that gilts fed 6 times/d had greater ADG than those fed the same amount over 2 feedings. To confirm these earlier responses, we used finishing pigs as a model in two 42-d trials and two 28-d trials to evaluate the effects of restricted feed intake and feeding frequency (2 vs. 6 times/d, floor fed) on pig performance between 68 and 114 kg. In all experiments, pigs (10/pen) were housed in 1.8 × 3.1 m pens with a half-solid, half-slatted concrete floor. Pigs were fed a corn- and soybean meal-based diet formulated to 1.15% standardized ileal digestible Lys and 3,294 kcal of ME/kg. In Exp. 1 to 3, energy and Lys were supplied to pigs according to NRC (1998) calculations to target an ADG of 0.80 kg. In Exp. 4, the diet was supplied to pigs to target an ADG of 0.80 kg (low feed intake) or 0.95 kg (high feed intake) to determine if the amount of energy above the maintenance requirement and feeding frequency affected pig performance. Pigs were fed by dropping similar amounts of feed onto the solid concrete floor either 2 (0700 or 1400 h) or 6 times (3 meals within 2 h at the morning and afternoon feedings) per day with an Accu-Drop Feed Dispenser (AP Systems, Assumption, IL). In Exp. 1 and 2, pigs fed 6 times daily had increased (P < 0.02) ADG and G:F compared with pigs fed 2 times per day. Greater feeding frequency increased (P < 0.05) the duration of time spent feeding and standing and reduced the lying time. In Exp. 3, a third treatment was included to determine whether the improvements in performance were due to decreased feed wastage. This treatment was designed to minimize feed wastage by dropping feed closer to the floor for pigs fed 2 times per day. Pigs fed 6 times daily had improved (P < 0.05) ADG and G:F compared with pigs in either treatment fed 2 times per day. No difference (P > 0.05) in performance was observed between pigs fed 2 times per day when feed was dropped from the feed drop or by the modified method. In Exp. 4, increasing the feeding frequency from 2 to 6 times per day improved (P < 0.01) ADG and G:F for pigs fed the low feed intake and tended to increase (P < 0.06) ADG and improved (P < 0.05) G:F for pigs fed the high feed intake. In limit-feeding situations, increasing the frequency of feeding from 2 to 6 times per day improved pig performance, which confirmed our earlier findings in gestating gilts.     

Endotoxin induces delayed ovulation following endocrine aberration during the proestrous phase in Holstein heifers

The effect of endotoxin on follicular growth and on secretion of LH, estradiol-17β, progesterone and cortisol during the proestrous phase in cattle was investigated. Holstein heifers were treated with PGF₂ at 11–13 d after ovulation to induce luteolysis. At 42 hr after PGF₂ treatment, heifers were administered either lipopolysaccharide (LPS; Escherichia coli, O111:B4, 5 μg/kg, n = 6) or saline (control; n = 6) by i.v. bolus injection. Ovarian structures were monitored daily by transrectal ultrasonography, and blood samples were collected at various times for hormonal analysis. The duration from PGF₂ treatment to ovulation was significantly longer in the LPS group (8.0 ± 1.3 d) than in the control group (4.2 ± 0.2 d). LPS significantly reduced the pulse frequency of LH for 6 hr after the administration, and increased the mean concentration and pulse amplitude of LH from 3 to 6 hr after the administration. The plasma concentrations of progesterone and cortisol were transiently increased after LPS administration. The plasma concentration of estradiol-17β was significantly decreased at 24 hr after LPS administration compared to that in the controls. Five of six LPS-treated heifers exhibited no preovulatory LH surge until 120 hr after PGF₂ treatment and the remaining heifer exhibited the surge at 108 hr after PGF₂ treatment, while the LH surge was observed at 54–78 hr after PGF₂ treatment in control heifers. These results suggest that endotoxin disrupts progression of the proestrous phase of cattle, interrupting the preovulatory estradiol rise and thus delaying the LH surge and the subsequent ovulation.     

Long-term feed intake regulation in sheep is mediated by opioid receptors

These experiments were conducted to determine if 1) syndyphalin-33 (SD33), a mu-opioid receptor ligand, affects feed intake; 2) SD33 effects on feed intake are mediated by actions on opioid receptors; and 3) its activity can counteract the reduction in feed intake associated with administration of bacterial endotoxin. In Exp. 1, 5 mixed-breed, castrate male sheep were housed indoors in individual pens. Animals had ad libitum access to water and concentrate feed. Saline (SAL; 0.9% NaCl) or SD33 (0.05 or 0.1 micromol/kg of BW) was injected i.v., and feed intake was determined at 2, 4, 6, 8, 24, and 48 h after the i.v. injections. Both doses of SD33 increased (at least P < 0.01) feed intake at 48 h relative to saline. In Exp. 2, SAL + SAL, SAL + SD33 (0.1 micromol/kg of BW), naloxone (NAL; 1 mg/kg of BW) + SAL, and NAL + SD33 were injected i.v. Food intake was determined as in Exp. 1. The SAL + SD33 treatment increased (P = 0.022) feed intake at 48 h relative to SAL + SAL. The NAL + SAL treatment reduced (at least P < 0.01) feed intake at 4, 6, 8, 24, and 48 h, whereas the combination of NAL and SD33 did not reduce feed intake at 24 (P = 0.969) or 48 h (P = 0.076) relative to the saline-treated sheep. In Exp. 3, sheep received 1 of 4 treatments: SAL + SAL, SAL + 0.1 micromol of SD33/kg of BW, 0.1 microg of lipopolysaccharide (LPS)/kg of BW + SAL, or LPS + SD33, and feed intake was monitored as in Exp. 1. Lipopolysaccharide suppressed cumulative feed intake for 48 h (P < 0.01) relative to saline control, but SD33 failed to reverse the reduction in feed intake during this period. These data indicate that SD33 increases feed intake in sheep after i.v. injection, and its effects are mediated via opioid receptors. However, the LPS-induced suppression in feed intake cannot be overcome by the opioid receptor ligand, SD33.     

Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs

In sheep, serum concentrations of leptin change congruently with increases or decreases in nutritional status, while intracerebroventricular infusions of leptin dramatically suppress feed intake in well-fed lambs, and may also increase growth hormone (GH), and/or luteinizing hormone (LH) in undernourished lambs. The objective of the present study was to determine the effects of peripherally delivered ovine leptin, via intravenous infusions, on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine. Twelve ewe lambs weighing 29.4 +/- 0.7 kg were infused intravenously with a linearly increasing dose of leptin or saline (n = 6 per group) for 10 days, reaching a maximum dose delivered of 0.5mg/h on day 10. Feed intake was assessed twice daily, and blood samples were collected every 10 min for 6 h on days 0, 2, 5, 8, and 10. Serum concentrations of leptin increased in leptin-treated lambs by day 2 (P = 0.05), and continued to increase to concentrations 9-fold greater than saline-infused lambs by day 10 (P < 0.001). Despite the substantial increase in serum leptin, feed intake did not differ between leptin and saline-infused lambs except on day 3.5 (P = 0.01). Furthermore, intravenous infusions of leptin did not significantly influence serum concentrations of insulin, cortisol, IGF-1, thyroxine, LH, or GH. Collectively, these observations contrast with the potent hypophagic effects of leptin when delivered intracerebroventricularly into well-fed lambs. The reasons for the disparate response of lambs treated intravenously with leptin, versus that reported for lambs treated intracerebroventricularly with leptin are not known, but may provide insight into the mechanism(s) of leptin resistance.     

Reproductive and growth responses of gilts to exogenous porcine pituitary growth hormone

Forty gilts (mean wt = 72 kg) were administered daily either vehicle (C = control) or 70 micrograms porcine growth hormone (pGH)/kg BW. After 30 d of treatment, eight gilts per group (Exp. 1) were slaughtered and blood, uteri and ovaries were collected. Follicular fluid (FFl) was collected and granulosa cells (GC) were cultured. The remaining gilts (Exp. 2) were treated for up to 35 additional days and examined twice daily for estrus. Estrusal gilts were removed from the experiment. Noncyclic gilts (n = 9 of 12 pGH; n = 4 of 12 C) were slaughtered on d 66 and their ovaries were examined. Ovarian weights were not different for pGH and C gilts in either Exp. 1 (P greater than .1) or Exp. 2 (P = .09). Uterine weights were greater for pGH-treated than for C gilts (P less than .007) in Exp. 1, but not in Exp. 2. Concentrations of estradiol (E2) in plasma and FF1 and of progesterone (P) in plasma and FF1 were not different for pGH and C gilts. Concentrations of insulin-like growth factor-I (IGF-I) in FF1 and in serum were greater for pGH than for C gilts (P less than .01). Concentration of P in serum-free medium of cultured GC was lower for GH than for C (P less than .05) in the presence or absence of gonadotropins in Exp. 1. The FSH-stimulated secretion of P was also lower for GC of pGH-treated gilts in Exp. 2, indicating a failure of GC to differentiate in culture. Only one pGH gilts in Exp. 2 manifested estrus, compared with seven C gilts (P less than .025). In Exp. 1, ADG was higher (P less than .03) and feed/gain lower (P less than .07) for pGH gilts. Longissimus muscle area (LMA) was not different (P = .19) between groups. Backfat thickness (BF) was lower (P less than .005) in pGH than in C in both Exp. 1 and 2. We conclude that exogenous pGH increased growth rate, improved feed efficiency and altered carcass traits in gilts. However, these effects were associated with impaired ovarian development of prepubertal gilts and a low incidence of estrus.     

The influence of growth hormone injections on the endocrine and metabolic status of gilts

Twelve Yorkshire x Landrace prepubertal gilts were assigned equally to treatments involving daily injections of either porcine growth hormone (GH, 90 micrograms/kg) or vehicle buffer from 150 to 159 d of age. Blood samples were obtained every hour from 0600 hr at 153 d until 0500 hr at 154 d of age, inclusively. At 0800 hr on 154 d, gilts received an injection of 500 IU PMSG, followed 96 hr later by 250 IU hCG. Gilts were slaughtered at 163 d and the ovaries recovered for an assessment of the ovarian response to the gonadotrophic stimulation. Five control gilts (83%) exhibited a normal ovulatory response but only one GH gilt (17%) was so designated (P less than 0.05). There was no apparent effect of treatment on serum concentrations of LH, FSH or cortisol. Growth hormone treatment reduced serum concentrations of T4 (P less than 0.001) and prolactin (P less than 0.02), but increased serum GH (P less than 0.001), T3 (P less than 0.06), insulin (P less than 0.001) and glucose (P less than 0.001). Serum concentrations of free fatty acids (FFA) were not significantly altered by exogenous GH. The concomitant elevation of serum insulin and glucose suggests that an insulin-resistant state was induced which, if evident at the ovarian level, may be a factor mediating the adverse effects of exogenous GH on ovarian function. The data presented also suggests that circulating concentrations of thyroid originating hormones are altered by exogenous GH.     

Influence of cumulative feed intake during early and mid-lactation on luteinizing hormone secretion and weaning-to-estrus interval in primiparous sows.

Twelve pregnant gilts were assigned to a completely randomized block design with two treatments in two blocks (2 farrowing groups). The treatments were a feeding amount of 6 kg or 2 kg/day provided during lactation. The lactation diet contained 18.6% crude protein, 1.0% lysine, and 3.27 Mcal/kg metabolizable energy (as-fed basis). Litters were weaned at 2100 on day 21 after farrowing. Blood samples for luteinizing hormone (LH) measurements were taken at 15-min intervals for 8 hr on day 12 of lactation, and samples for glucose and insulin were collected at 1-hr intervals for 3 hr on day 12. The effects of feed intake treatments on LH pulse frequencies (2.9 vs 0.7) and insulin concentrations (15.0 vs 8.9 IU/mL) were found (P < 0.05) on day 12 of lactation. In regression analysis, greater cumulative feed intake from 1 to 12 days was associated with higher insulin concentrations (P = 0.04), greater LH pulse frequencies (P = 0.01) on day 12 of lactation, and shorter weaning-to-estrus intervals (WEI) (P = 0.03). Furthermore, an association between insulin concentrations and LH pulse frequencies was found on day 12 of lactation (P = 0.01). Using regression models for weaning-to-estrus interval, when each cumulative feed intake from 4 to 21 days was used as an independent variable, the R2 values increased from 0.24 to 0.37. These results suggest that feed intake during early and mid-lactation influences LH secretion as early as day 12 after farrowing, and is associated with shorter WEI. This research also indicates that feed intake from 4 to 12 days of lactation is more important than that during the first few days after farrowing.     

The effect of space allocation on barrow and gilt performance

Two experiments were conducted to determine the variation in response to space allocation between barrows and gilts and to examine an alternative allocation regimen for barrows and gilts. Experimental space allocations in both experiments were achieved by varying the number of pigs per pen in a fully slatted facility. In Exp. 1, barrows were given 0.58 and 0.65 m2/pig (nine and eight pigs per pen, respectively) and gilts were given 0.65 and 0.74 m2/pig (eight and seven pigs per pen, respectively). In addition, barrows at 0.58 m2/pig were fed diets formulated for barrows or diets formulated for gilts. Barrows grew 4.8% slower (P = 0.031) and ate 3.1% less feed daily (P = 0.062) at 0.58 vs. 0.65 m2/pig from 22 to 115 kg BW, with no difference in feed conversion, daily lean gain, carcass lean percent, or variation in weight within the pen at time of first pig removal to slaughter. There was no improvement in daily gain, feed intake, feed efficiency, lean gain, or carcass lean percent when gilts were given 0.74 vs. 0.65 m2/pig from 22 to 115 kg BW. There was no difference in performance between the population that consisted of barrows and gilts at 0.65 m2/pig vs. the population of barrows at 0.58 m2/pig and gilts at 0.74 m2/pig. There was no difference in performance by barrows at 0.58 m2/pig when fed either barrow or gilt diets, except for a slight increase (P = 0.078) in within-pen weight variation when the first pig was removed for slaughter for the barrows fed gilt diets. In Exp. 2, barrows and gilts were given 0.58 m2/pig or 0.74 m2/pig (18 vs. 14 pigs per pen) from weaning (mean age 17 d) to slaughter on d 168 postweaning. There were no interactions between space allocation and gender. Daily gain and feed intake were decreased by 2.8% (P = 0.037) and 2.9% (P = 0.084), respectively, with no effect on feed conversion or standardized fat-free lean daily gain for the 0.58 vs. the 0.74 m2/pig treatment, whereas total live weight gain per pen was increased 20.8% (P < 0.001). Results of Exp. 1 suggest that space allocation can be used to achieve similar growth rates between barrows and gilts, and results of Exp. 2 suggest that the response to space allocation is similar for barrows and gilts. The difference in magnitude of response to space allocation between experiments may be due in part to when the social group was formed, with a smaller difference in performance in Exp. 2 associated with a stable social group from weaning to slaughter.     

Modulation of growth hormone-releasing factor-induced release of growth hormone from bovine pituitary cells

Growth hormone (GH)-releasing factor (GRF) at concentrations of 10−¹² through 10−⁷M for 6 hr linearly increased GH release (b₁ = 10.4 ± .3) from bovine anterior pituitary cells in culture. Maximum release of GH (262% above controls) occurred at 10−⁷M GRF. In contrast, GH release-inhibiting factor (SRIF) at 10−¹² through 10−⁵M had no effect on basal concentrations of GH. In a second experiment, as the proportion of SRIF relative to GRF increased. SRIF suppression of GRF-induced GH release from anterior pituitary cells increased. In a third experiment, anterior pituitary cells cultured in media containing fetal calf serum (FCS) were treated with cortisol (0 or 10 ng/ml media) for 24 hr before exposure to 10−¹³ through 10−⁷M GRF. GRF linearly increased GH secretion (b₁ = 7.4 ± .3) and cortisol augmented this response (b₁ = 10.5 ± .6). However, when cells were cultured in media containing dextran-charcoal treated FCS, cortisol did not alter GRF-induced GH release. Our results demonstrate that GH response of bovine anterior pituitary cells to GRF was modulated negatively by SRIF. However, augmentation of GRF-induced GH release by cortisol was evident only when cells were cultured in media supplemented with untreated FCS.     

Luteinizing hormone secretion in hypophysial stalk-transected gilts given hydrocortisone acetate and pulsatile gonadotropin-releasing hormone.

The site within the hypothalamic-pituitary axis at which cortisol acts to inhibit luteinizing hormone (LH) secretion was investigated in female pigs. Six ovariectomized, hypophysial stalk-transected (HST) gilts were given 1 microgram pulses of gonadotropin releasing-hormone (GnRH) iv every 45 min from day 0 to 12. On days 6-12, each of 3 gilts received either hydrocortisone acetate (HCA; 3.2 mg/kg body weight) or oil vehicle im at 12-hr intervals. Four ovariectomized, pituitary stalk-intact gilts served as controls and received HCA and pulses of 3.5% sodium citrate. Jugular blood was sampled daily and every 15 min for 5 hr on days 5 and 12. Treatment with HCA decreased serum LH concentrations and LH pulse frequency in stalk-intact animals. In contrast, serum LH concentrations, as well as the frequency and amplitude of LH pulses, were unaffected by HCA in HST gilts and were similar to those observed in oil-treated HST gilts. We suggest that chronically elevated concentrations of circulating cortisol inhibit LH secretion in pigs by acting at the level of the hypothalamus.