Effect of Naloxone on Serum Luteinizing Hormone Concentration in Anovulatory Holstein Cows During the Early Postpartum Period

This experiment was conducted to investigate the effect of naloxone (NAL), an opioid receptor antagonist, on pituitary LH secretion in anovulatory Holstein cows during the early postpartum period when cows were expected to be in negative net energy balance. Twenty-three cows (11 primiparous) received either saline (n = 12) or 1 mg/kg BW NAL i.v. (n = 11) on Day 14 or 15 postpartum. Jugular blood samples were collected at 15-min intervals for 2 hr before and 2.5 hr after NAL or saline. All cows received 3 ug gonadotropin releasing hormone (GnRH) at 2.5 hr post-NAL or -saline and blood collection was continued for 1 hr. Mean serum progesterone concentration was 0.33 ± 0.2 ng/ml. Mean net energy balance for all cows was -5.5 ± 0.6 Mcal/day. Naloxone caused a transient increase (P < 0.05) in serum LH concentrations in both primi- and multiparous cows within 45 min after administration. In contrast, serum LH concentrations remained unchanged in saline-treated cows. GnRH increased (P < 0.05) LH and there was no effect of treatment. These results suggest that modulation of LH secretion, at least in part, may be mediated via endogenous opioids in dairy cows before first postpartum ovulation.     

Postpartum reproduction in protein restricted beef cows: effect on the hypothalamic-pituitary-ovarian axis

The influence of dietary CP on circulating LH and anterior pituitary and hypothalamic function was examined. In Exp. 1, 28 cows were randomly assigned to four treatment groups: adequate CP (ADQ; .96 kg/d) or deficient CP (DEF; .32 kg/d) beginning at 90, 60 and 30 d before parturition and continued at a 33% increase in feed consumption after parturition. Cows were bled at 15-min intervals for 8 h on d 20, 40 and 60 after parturition. Pituitaries were collected on d 62 to analyze GnRH receptor numbers and gonadotropin content. Frequency of pulsatile LH release increased (P less than .05) from 20 to 60 d in ADQ cows. Basal and mean LH were not affected (P greater than .10) by CP restriction or by days after parturition. Crude protein did not affect pituitary GnRH receptors (P greater than .10), but it did affect pituitary LH content, FSH content and FSH concentration (P less than .05). In Exp. 2, 28 cows were assigned to treatment groups as in Exp. 1. All cows were challenged with GnRH (.22 micrograms/kg BW) at 20, 40 and 60 d after parturition and were bled every 30 min for 6 h. Responsiveness to GnRH increased with increased time after parturition (P less than .07). Deficient CP decreased GnRH-induced LH release (P less than .05). In Exp. 3, 12 cows were randomly assigned to ADQ or DEF CP beginning 120 d before parturition. All cows received 1 mg estradiol-17 beta (E2) on d 19, 39 and 59 after parturition and were bled every 30 min for 14 h beginning 14 h following E2. Response to E2 was unaffected by CP restriction (P greater than .10), whereas time to E2-induced LH peak decreased as time after parturition increased in ADQ cows (P less than .05). Results suggest that delayed return to estrus in CP-deficient postpartum beef cows might be due to reduced gonadotropin release from the anterior pituitary and decreased anterior pituitary responsiveness to GnRH.     

Effects of progesterone and weaning on LH and FSH responses to naloxone in postpartum beef cows

Two experiments were conducted to evaluate the effects of naloxone, an endogenous opioid receptor antagonist, on LH and FSH secretion in postpartum beef cows. In Experiment 1, 24 cows were divided into three equal groups. On day 15 postpartum, all cows were bled for 8 hr at 10 min intervals to evaluate LH secretory parameters. On day 18 postpartum, three treatments were administered: (a) saline at 0730 and 1130 hr; (b) 275 mg naloxone at 0730 and 1130 hr; (c) naloxone as in (b) above, plus this group was also treated with 50 mg progesterone (P4) twice daily from day 16 to day 19. In each treatment, jugular vein samples were collected at 10 min intervals from 0800 to 1600 hr. On day 19 the same treatments were administered at the same times, however, all cows were given 25 micrograms GnRH at 1200 hr to evaluate the LH secretory response. Naloxone increased mean LH concentration (P less than .05) and tended to increase pulse amplitude and frequency compared to controls. However, the most dramatic difference was due to P4 treatment which suppressed mean LH, pulse amplitude and frequency. Treatments had no effect on LH secretion in response to a 25 micrograms dose of GnRH. In Experiment 2, the effects of suckling on the naloxone response were examined in 16 postpartum cows. On day 21 postpartum, blood was collected at 10 min intervals for 8 hr and then calves were removed from half the cows. After 3 days of calf removal, all cows were sampled at 10 min intervals for 4 hr; then naloxone was injected after each 10 min sample at a dose rate of 200 mg/hr (33 mg per injection). Naloxone treatment and sampling continued for an additional 8 hr. Calf removal alone had very little effect on LH pulsatility. However, naloxone resulted in increased pulse frequency and mean LH compared to the control period. We conclude that LH release in the early postpartum cow is partially regulated by endogenous opioid peptides. We were unable to detect any effects on FSH secretion nor on pituitary sensitivity to exogenous GnRH.     

The change of dry matter intake, milk yield and bone turnover in primiparous cows compared to multiparous cows during early lactation

Data from six primiparous and nine multiparous Holstein cows were used to clarify the difference of Ca and P mobilization between primiparous and multiparous cows during early lactation. The dry matter intake (DMI) of primiparous cows was lower (P < 0.01) than those of multiparous cows. Milk yield was lower in primiparous cows at 7, 14, 21 (P < 0.01), and 28 days (P < 0.05) after parturition. There was no significant difference in milk Ca and P concentrations between primiparous and multiparous cows. There were no significant differences in plasma Ca and P concentrations between primiparous and multiparous cows. The plasma P level at 7 days postpartum in primiparous cows was lower (P < 0.05) than 28 days postpartum. The concentration of plasma osteocalcin (OC) measured as bone formation marker of primiparous cows was significantly higher than multiparous cows (P < 0.01) at 21 and 28 days postpartum. The urinary deoxypyridinoline (DPD) as bone resorption marker of primiparous cows tended to be higher (P < 0.10) than multiparous cows at 21 days after parturition and decreased to the same level as that of multiparae toward the peak lactation. These results show that Ca and P mobilization of primiparous cows are more active than multiparous cows.     

Effect of copper status, supplementation, and source on pituitary responsiveness to exogenous gonadotropin-releasing hormone in ovariectomized beef cows

The effect of Cu status, supplementation, and source on pituitary responsiveness to exogenous GnRH was evaluated using nine multiparous, nonpregnant, nonsuckling, ovariectomized Angus cows (7.1 +/- 3.3 yr; 622.9 +/- 49.8 kg; BCS = 6.0 +/- 0.5). Cows were considered Cu-deficient based on liver Cu concentrations (< 30 mg of Cu/kg of DM) after receiving a low-Cu, forage-based diet supplemented (DM basis) with 5 mg of Mo/kg and 0.3% S for 216 d. Copper-deficient cows were stratified based on age, BW, BCS, and liver Cu concentration and assigned randomly to repletion-phase treatments. Treatments included 1) control (no supplemental Cu); 2) organic (ORG; 100% organic Cu); and 3) inorganic (ING; 100% inorganic CuSO4). Treatments were formulated to meet all NRC recommendations, except for Cu, which was supplemented to ORG and ING cows at 10 mg of Cu/kg of dietary DM. During the 159-d repletion phase, Cu status was monitored via liver biopsy samples, and all cows received exogenous progesterone. A controlled intravaginal drug-release device (replaced every 14 d) was used to maintain luteal phase progesterone as a means to provide negative feedback on the hypothalamic-pituitary axis. During the repletion phase, liver Cu concentrations did not differ between ORG and ING cows at any time. By d 77 of the repletion phase, all supplemented cows were considered adequate in Cu, and liver Cu concentrations were greater in supplemented than in nonsupplemented control cows on d 77 (P < 0.05) and throughout (P < 0.01) the repletion phase. Beginning on d 99, exogenous GnRH was administered to all cows at low (0, 3, and 9 microg; Exp. 1) and high doses (0, 27, and 81 microg; Exp. 2) at six different times. Cows were catheterized every fifth day, and blood samples were collected every 15 min for 1 h before and 4 h after GnRH administration and analyzed for LH concentration. In Exp. 1, Cu status and supplementation did not affect basal or peak LH concentrations, but total LH released tended (P < 0.07) to be greater in Cu-supplemented vs. control cows when 3 microg of GnRH was administered. In Exp. 2, there was no effect of Cu supplementation or source on basal, peak, or total LH released, regardless of GnRH dose. Pituitary LH concentrations did not differ across treatments. In conclusion, Cu status, supplementation, and source did not affect GnRH-induced LH secretion or pituitary LH stores in ovariectomized, progesterone-supplemented cows in this experiment.     

Effect of metoclopramide on luteinizing hormone secretion in postpartum anestrous cows.

The effect of metoclopramide (MC), a dopamine antagonist on luteinizing hormone (LH), was examined in anestrous primaparous cows. Metoclopramide has been found to be beneficial in overcoming fescue toxicosis; increasing LH secretion stimulates return to ovulatory function after parturition. Consequently, if MC had negative effect on LH secretion, it would indicate that administration of MC to reproducing animals might be limited. Of 14 postpartum (47 to 66 days) cows, 7 were given MC (4 mg/kg of body weight, IV), and 7 served as controls. Blood was obtained via jugular cannulas at 15-minute intervals for 8 hours; MC was given at the end of the first hour, and gonadotropin-releasing hormone (GnRH, 7 mg/kg), was given IV at the end of hour 7 as a challenge stimulus for LH secretion. Prior to GnRH administration, MC did not have significant effect on LH secretion, as judged by mean serum LH concentration, LH pulse frequency, and LH pulse amplitude. Administration of MC resulted in greater (P less than 0.05) LH response to GnRH, indicating enhanced secretory ability when the pituitary gland was challenged. Serum prolactin concentration was increased (P less than 0.01) by MC administration. Therefore, MC did not have adverse effect on LH secretion in postpartum cows.     

Gonadotropin releasing hormone-induced secretion of luteinizing hormone during the milk-ejection reflex in the postpartum beef cow

A study was conducted to determine the effect of the milk-ejection reflex on exogenous gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH) after short-term calf removal. Twenty-four postpartum multiparous beef cows were assigned randomly to groups arranged in a 2(3) factorial arrangement. Factors consisted of two levels of suckling [suckled (S) or nonsuckled (NS)], treatment with GnRH [saline (C) or 200 micrograms GnRH] and days postpartum (d 1 and 14). Dams were isolated from their calves for 4 h on d 1 and 14 postpartum. At the end of 4 h dams were reunited with their calves in S + C and S + GnRH groups, while dams of calves in NS + C and NS + GnRH groups remained separated an additional 2 h. Cows were injected iv with saline or GnRH following the 4-h isolation period, 5 min after calves had begun suckling or nuzzling the udder. Sera from jugular blood samples collected 15 min prior to the end of the 4-h isolation period, immediately prior to injection (0 h) and at 15 min intervals thereafter for 120 min were analyzed for LH. Serum concentrations of LH in control cows did not differ due to suckling or stage of the postpartum period and averaged 2.3 +/- .1 ng/ml. Pituitary response to GnRH was determined by computing the rate of LH release. Rate of LH release (ng LH.ml-1.min-1) in response to GnRH on d 14 was greater (P less than .001) than on d 1 in both suckled and nonsuckled groups (S + GnRH, 37.1 +/- 3.9 vs 18.3 +/- 5.0; NS + GnRH, 34.7 +/- 5.9 vs 14.5 +/- 1.1). However, GnRH-induced release of LH did not differ between suckled and nonsuckled cows on either d 1 or 14 postpartum. These data indicate that response of the bovine pituitary to GnRH during the postpartum period is not influenced by the act of suckling but is enhanced with time after parturition.     

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 +/- 3.2 ng) and HYP (14.0 +/- 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 +/- .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 +/- .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 +/- .2 pmoles/pituitary) and estradiol (5.0 +/- .9/moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.     

GnRH in the infundibular stalk-median eminence is related to percentage body fat in carcasses of beef cows.

Mature Hereford cows (n = 28) were used to determine the effect of percentage body fat on secretion of LH and content of GnRH in the infundibular stalk-median eminence (ISME). Cows were fed to maintain, lose, or gain weight to achieve body condition scores (BCS; 1 = emaciated; 9 = obese) of 3 to 7. Then cows were fed to maintain weight and body condition. Before slaughter, estrus was synchronized using two injections of prostaglandin F2 alpha(PGF) 11 d apart. Five d after the second PGF injection, cows were given 100 micrograms of GnRH (im) and serum samples were obtained. LH was quantified using RIA. The anterior pituitary and ISME were obtained within 45 min of death. Anterior pituitary weight and LH concentration, total GnRH in the ISME, total carcass fat, and percentage carcass fat were determined. BCS of cows at the time of slaughter influenced percentage carcass fat (P less than .001), total GnRH in the ISME (P less than .02), and maximum LH after GnRH treatment (P less than .09), but did not influence pituitary weight or concentration of LH in the pituitary. Content of GnRH in the ISME averaged 76 +/- 12, 32 +/- 14, 27 +/- 13, and 24 +/- 13 ng for cows with BCS of 3, 5, 6, and 7, respectively. BCS was correlated (P less than .001) with percentage carcass fat (r = .94) and total fat in the carcass (r = .92). Total GnRH in the ISME was negatively correlated (P less than .005) with BCS (r = -.54), percentage carcass fat (r = -.55), and total carcass fat (r = -.49).(ABSTRACT TRUNCATED AT 250 WORDS)     

Postpartum nutrient intake and body condition: effect on pituitary function and onset of estrus in beef cattle

From calving through first estrus, 30 Brangus females were assigned equally to one of three diets to study the effect of postpartum nutrient intake and body condition on the ability of the pituitary to release luteinizing hormone (LH) and on the postpartum interval to estrus (PPI). The postpartum diets were calculated to achieve a 1) low [90% of the National Research Council (NRC) recommendations], 2) maintenance (100% of the NRC recommendations), or 3) high (110% of the NRC recommendations) level of nutrient intake. The females were group-fed within a treatment and calves were allowed to suckle ad libitum. Cow weight, body condition score and calf weight were recorded 24 h aftercalving, d 20 postpartum and at first behavioral estrus. On d 21 postpartum, blood samples were collected via jugular cannulae at 15-min intervals for 4 h, followed by a 100-micrograms im injection of gonadotropin-releasing hormone (GnRH) and continued sampling at 15-min intervals for an additional 6 h to determine serum LH. Although there was a significant decrease in PPI with increasing levels of nutrient intake (low = 57.5 +/- 8.8 d; maintenance = 40.3 +/- 6.6 d; high = 34.7 +/- 5.1 d), there were no differences in any of the observed LH characteristics (or variables) due to treatment. There were, however, marked differences in both the PPI and LH characteristics when data were analyzed on the basis of ability to maintain body condition from calving through 20 d postpartum, regardless of calculated dietary treatments. Cows that maintained body condition (MBC) had a shorter PPI [MBC, 31.7 +/- 2.8 vs lost (LBC) 60.0 +/- 7.5 d; P less than .01], higher basal levels of endogenous LH (MBC, .83 +/- .09 vs LBC, .61 +/- .04 ng/ml; P less than .025), higher GnRH-induced peak LH concentration (MBC, 58.99 +/- 11.15 vs LBC, 38.86 +/- 8.37 ng/ml. P less than .10), higher LH levels throughout the GnRH-induced LH surge (P less than .001), and greater release curve areas for the endogenous (MBC, 124.6 +/- 13.3 vs LBC, 91.7 +/- 5.6 units; P less than .025), GnRH-induced (MBC, 4370.8 +/- 699.5 vs LBC, 3039.7 +/- 683.3 units; P less than .10) and total (MBC, 4510.7 +/- 706.7 vs LBC, 3141.9 +/- 684.7 units; P less than .10) LH release.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cortisol and luteinizing hormone after adrenocorticotropic hormone administration to postpartum beef cows

Cortisol and luteinizing hormone (LH) were measured in serum after the administration of adrenocorticotropic hormone (ACTH) to suckled (S) and nonsuckled (NS) beef cows. Blood was sampled on 2 consecutive days every 2 weeks for four bleeding periods starting 14 days after calving. Cows were injected with 200 IU ACTH or saline in a 2-day switchback design. Serum was collected before ACTH or saline injection and at 30-min intervals thereafter for 8 hours. Average cortisol concentrations in serum were similar in S and NS cows (6.4 +/- .6 and 6.1 +/- .8 ng/ml, respectively) after saline. Average cortisol concentrations in serum collected during an 8-hr period after ACTH on days 14, 28, 42 and 56 postpartum were 24.7 +/- 2.4, 31.8 +/- 3.5, 36.4 +/- 4.2 and 40.7 +/- .5 ng/ml, respectively, for S cows, and 31.1 +/- 2.9, 44.7 +/- 5.2, 45.0 +/- 5.7 and 46.0 +/- 5.4 ng/ml, respectively, for NS cows. Cortisol response to ACTH, measured as area under the response curve, was greater (P less than .05) in NS than in S cows. Amount of cortisol released by 200 IU ACTH was maximal by days 28 to 29 postpartum in NS cows, but the response increased gradually between days 14 to 15 and days 56 to 57 in S cows. overall, LH in serum averaged .55 +/- .08 ng/ml for S cows and .92 +/- .06 ng/ml for NS cows after saline, and .49 +/- .07 ng/ml for S cows and .94 +/- .06 ng/ml for NS cows after ACth. Although mean and peak serum LH concentrations did not differ between cows given ACTH and those given saline, the number of LH peaks and the number of cows having LH after saline. Mean serum LH concentrations were lower (P less than. 05) in S than in NS cows at 28 days postpartum. The number of LH peaks was lower (P less than .05) and the magnitude of the largest LH peak tended to be lower (P less than .06) in S cows at all sampling periods.     

Synchronization of estrus in postpartum beef cows and virgin heifers using combinations of melengestrol acetate, GnRH, and PGF2alpha

The efficacy of various combinations of melengestrol acetate (MGA), GnRH, and PGF2alpha for the synchronization of estrus in Angus-based beef cattle was compared. Hormones were administered as follows: MGA, 0.5 mg x animal(-1) x d(-1) mixed in a grain carrier; GnRH, 100 microg i.m.; PGF2alpha, 25 mg i.m. In Exp. 1, 2, and 3, cows were randomly assigned to treatments by parity and interval postpartum. The detection of estrus and AI were conducted from d -2 until 72 to 96 h after PGF2alpha, at which time cows not detected to be in estrus received GnRH and fixed-time AI (TAI). Data were analyzed separately for primiparous and multiparous cows. In Exp. 1, cows (n = 799) at three locations received GnRH on d -7 and PGF2alpha on d 0 and either no further treatment (GnRH-PGF) or short-term MGA from d -6 through d -1 (STMGA). Among multiparous cows, conception rate at TAI was greater (P < 0.05) for STMGA (41%, 47/115) than for GnRH-PGF treated cows (26%, 24/92). Across herds and parity, synchronized AI pregnancy rate (SPR) was not affected (P > 0.10) by treatment (GnRH-PGF vs. STMGA; 54%, 210/389 vs. 57%, 228/402). In Exp. 2, cows (n = 484) at three locations received either STMGA or long-term MGA from d -32 through d -19, GnRH on d -7, and PGF2alpha on d 0 (LTMGA). Among primiparous cows, SPR was greater (P < 0.01) in LTMGA (65%, 55/85) than STMGA-treated cows (46%, 40/87). Treatment had no effect (P > 0.10) on SPR among multiparous cows (STMGA vs. LTMGA; 59%, 92/155 vs. 64%, 101/157). In Exp. 3, cows (n = 838) at four locations received the LTMGA treatment and either no further treatment or an additional period of MGA exposure from d -6 through d -1 (L&STMGA). Among primiparous cows, SPR tended to be influenced (P < 0.10) by the herd x treatment interaction and was greater (P < 0.01) among L&STMGA (86%, 19/22) than LTMGA-treated cows (56%, 14/25) at a single location. Among multiparous cows, SPR was lower (P < 0.05) in L&STMGA (46%, 165/358) than LTMGA-treated cows (55%, 184/336). In Exp. 4, Angus heifers (n = 155) received either STMGA or 14 d of MGA (d -32 through d -19) and PGF2alpha on d 0 (MGA-PGF). The detection of estrus and AI were conducted from d -2 to d 6. Interval to estrus was greater (P < 0.05) and estrous response was lower (P < 0.05) in STMGA than MGA-PGF-treated heifers. In conclusion, primiparous cows responded more favorably to longer-duration MGA treatments than did multiparous cows. All protocols achieved sufficient SPR to justify their use for improved reproductive management of postpartum beef cows.     

Follicular, hormonal, and pregnancy responses of early postpartum suckled beef cows to GnRH, norgestomet, and prostaglandin F2alpha.

ABSTRACT: Cycling (n = 16) and noncycling (n = 24), early postpartum, suckled beef cows of three breeds were assigned randomly to three treatments: 1) 100-microg injection of GnRH plus a 6-mg implant of norgestomet administered on d -7 before 25 mg of PGF2alpha and implant removal on d 0 (GnRH+NORG); 2) 100 microg of GnRH given on d -7 followed by 25 mg of PGF2alpha on d 0 (GnRH); or 3) 2 mL of saline plus a 6-mg implant of norgestomet administered on d -7 followed by 25 mg of PGF2, and implant removal on d 0 (NORG). All cows were given 100 microg of GnRH on d +2 (48 h after PGF2alpha). Blood sera collected daily from d -7 to d +4 were analyzed for progesterone and estradiol-17beta, and ovaries were monitored daily by transrectal ultrasonography to assess changes in ovarian structures. Luteal structures were induced in 75% of noncycling cows in both treatments after GnRH, resulting in elevated (P < .01) progesterone on d 0 for GnRH+NORG-treated cows. Concentrations of estradiol-17beta (P < .01) and LH (P < .05) were greater on d +2 after GnRH for cows previously receiving norgestomet implants. Pregnancy rates after one fixed-time AI at 16 h after GnRH (d +2) were greater (P < .05) in GnRH+NORG (71%) than in GnRH (31%) and NORG (15%) cows. Difference in pregnancy rate was due partly to normal luteal activity after AI in over 87% of GnRH+NORG cows and no incidence of short luteal phases. The GnRH+NORG treatment initially induced ovulation or turnover of the largest follicle, induction of a new follicular wave, followed later by increased concentrations of estradiol-17beta and progesterone. After PGF2alpha, greater GnRH-induced release of LH occurred in GnRH+NORG cows before ovulation, and pregnancy rates were greater after a fixed-time AI.     

Alteration of the GnRH-induced LH release by steroids in postpartum dairy cattle

The effect of elevated plasma concentrations of estradiol-17 beta (E2 beta), estrone (E1) and progesterone (P), in concentrations similar to those observed at the end of pregnancy, on the gonadotropin releasing hormone (GnRH)-induced luteinizing hormone (LH) release in postpartum dairy cows was studied. Twenty-five dairy cows in late gestation were assigned to five groups of five each to receive daily steroid treatments as follows: 1 and 2) no exogenous steroids; 3) 20 mg E2 beta and 30 mg E1; 4) 150 mg P and 5) 20 mg E2 beta + 30 mg E1 + 150 mg P. Steroids were dissolved in alcohol (vehicle) and injected sc twice daily. Cows receiving no steroids were given vehicle. Administration of steroids or vehicle began immediately after parturition (d 0) and continued for 7 d to maintain concentrations of steroids in plasma similar to prepartum concentrations. Cows in groups 2 through 5 received an injection of 100 micrograms GnRH on d 2, 8, 16, 24 and 32 postpartum, while those in group 1 received water (vehicle for GnRH) on the same days. Plasma for hormonal determinations was collected on alternate days beginning 10 d before the expected day of parturition, daily through the period of steroid treatments (d 0 to 6, postpartum) and on alternate days thereafter until d 40 postpartum. In addition, plasma was collected immediately before GnRH or water administration and at .5 h intervals thereafter for 4 h. Trends in response to treatment over days postpartum were studied by partitioning sums of squares due to linear, quadratic and cubic polynomial responses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary concentrations of gonadotropins and receptors for GnRH in suckled beef cows at various intervals after calving

Mature beef cows were slaughtered at 5 (n = 6), 10 (n = 6), 20 (n = 6) or 30 (n = 5) d after calving to identify endocrine events that may affect the duration of postpartum anestrus. Additional cows (n = 6) were slaughtered 12 to 14 d after their first postpartum estrus (luteal phase cows). Anterior pituitary concentrations of luteinizing hormone (LH) were low at d 5 (383 +/- 69 micrograms/g), averaged 445 +/- 103 and 682 +/- 207 micrograms/g at d 10 and 20, respectively, and were elevated (P less than .05) by d 30 (1,097 +/- 174 micrograms) to a concentration similar to luteal phase cows (1,208 +/- 148 micrograms/g). Concentrations of follicle-stimulating hormone (FSH) averaged 12.4 +/- 1.1, 9.6 +/- 2, 8.6 +/- 1.8 and 7.4 +/- 3.3 mg/g at d 5, 10, 20 and 30, respectively. Affinity (1.6 +/- .2 X 10(9) M-1) of anterior pituitary receptors for the GnRH (gonadotropin-releasing hormone) analog (DAla6; des-Gly10, [D-Ala6]-LH-RH ethylamide) and weights (2.1 +/- .1 g) of the anterior pituitaries did not differ among groups (P greater than .05). Number of receptors for GnRH averaged 37 +/- 7, 39 +/- 9, 25 +/- 5 and 23 +/- 5 X 10(-14) M/mg protein at d 5, 10, 20 and 30, respectively. Anterior pituitaries from luteal phase cows contained 22 +/- 2 X 10(-14) M/mg protein of receptors for GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of GnRH pretreatment on reproductive performance of postpartum suckled beef cows following synchronization of estrus using GnRH and PGF2alpha

The effect of GnRH pretreatment on estrus detection rate, precision of estrus, and reproductive performance of postpartum beef cows synchronized to estrus using GnRH and PGF2alpha was evaluated. In Exp. 1, Angus cows (n = 87) were randomly assigned by parity, postpartum interval, and body condition score (BCS) to receive either 1) GnRH on d -7 and PGF2alpha on d 0 (GP) or 2) the GP treatment and an additional injection of GnRH on d -16 (GGP). Estrus detection and AI were conducted twice daily from d -3 to d 3. At 72 h after PGF2alpha, all animals not previously detected in estrus were bred by AI and received a concurrent injection of GnRH (TAI). Synchronized pregnancy rates were numerically increased (P = 0.15) in cows treated with GGP (55%) compared with those on the GP treatment (44%). In Exp. 2, 1,276 spring-calving, suckled beef cows in nine herds were randomized to treatments as described for Exp. 1, except that the initial GnRH injection for the GGP treatment was administered on d -14. Herd affected all indicators of reproductive performance (P < 0.05). The percentage of animals detected in estrus prematurely (d -3 to d 0; 7%) was not affected by treatment. Estrus response rate was influenced by postpartum interval (< 60 vs > or = 60; 61 vs 73%; P < 0.01) and a three-way interaction of parity, BCS, and treatment (P < 0.01). Within animals with a BCS > or = 5.5, the GGP treatment tended to increase the detection of estrus in primiparous cows (GP vs GGP; 76 vs 91%; P = 0.11) and decrease detection in multiparous cows (GP vs GGP; 78 vs 72%; P < 0.10). However, because conception rate to TAI in animals with a BCS > or = 5.5 was greater (P < 0.05) in the GGP than in the GP group (28 vs 8%, respectively), this interaction was interpreted to represent a shift in interval to estrus induced by the GGP treatment, rather than a reduction in the synchronization of ovarian function. Conception rates of animals inseminated to an observed estrus did not differ among treatments (P = 0.15). Synchronized pregnancy rate tended (P = 0.06) to be greater in GGP- (53%) than in GP-treated animals (47%). In conclusion, pretreatment with GnRH tended to increase pregnancy rates during a 6-d synchronization period, primarily through enhanced conception rates of cows bred by TAI. In contrast to our hypothesis, GnRH pretreatment did not increase the percentage of animals detected in estrus or the precision of estrus expression.     

Postpartum interval in beef cows shortened by enclomiphene

This study was designed to determine whether an anti-estrogen can block the negative effect of estrogens on luteinizing hormone (LH) release and therefore decrease the postpartum interval in suckled beef cows. In Exp. I, eight suckled postpartum beef cows were randomly assigned to treatment and control groups. Treatment cows received 1 g/d clomiphene citrate (im) from d 21 to 28 postpartum, while control cows were injected with saline. On d 28 postpartum, there was no difference (P greater than .05) in mean total and basal LH concentrations or LH pulse frequency between treatment and control cows. All control cows exhibited estrus on d 52 +/- 3; treatment cows exhibited estrus on d 134 +/- 12 (P less than .05). In Exp. II, 17 suckled cows were randomly assigned to three treatment groups: 1) control group (n = 6) receiving one empty implant, 2) 10-cm enclomiphene implant group (n = 5) and 3) 30-cm enclomiphene implant group (n = 6). The silastic implants were placed sc on d 20 and removed on d 29 postpartum. Mean total LH concentrations during d 24 to 29 postpartum in the 30-cm enclomiphene implant group were higher than the 10-cm implant (P less than .05) and control group (P less than .05). The postpartum period in the 30-cm enclomiphene group (45 +/- 6 d) was shorter than the 10-cm implant (94 +/- 24 d) and control (96 +/- 20 d) groups (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Response of postpartum beef cows to exogenous progestogens and gonadotropin releasing hormone

Plasma progesterone (P4) profile and estrous detection were used during three experiments to evaluate the effects of exogenous progestogens on the life span of gonadotropin releasing hormone (GnRH)-induced corpora lutea (CL) in postpartum (pp) beef cows. Experiment 1 utilized primiparous fall-calving cows (n = 28, trial 1); and spring-calving cows (n = 29, trial 2). On d 18 to 27 pp (d 0) all cows received intravaginal devices containing either P4 or no P4 (NP) for 5 d. On d 5 the devices were removed and calves were either removed (CR) or were present (CP) with half of the cows within steroid group. At 50 h after device removal, 500 micrograms of GnRH was given (iv) to all cows, and weaned calves were reunited with their dams. The induced CL had a normal life span (greater than 16 d) in 17 and 86% (trial 1) and 8 and 79% (trial 2) of NP and P4 cows, respectively. Calf removal did not affect (P greater than .10) the life span of the CL. In Exp. 2, spring-calving multiparous cows (d 18 to 24 pp; d 0) received either no P4 (NP; n = 19), P4 for 6 d via intravaginal devices (P4H; n = 19) or a single im injection of 300 mg P4 (P4 IM; n = 18). At 48 h after device removal or at 8 d after the injection of P4, half of the cows within steroid group received either 500 micrograms GnRH or saline. Corpora lutea had a normal life span in 0, 11, and 80% of NP, P4 IM and P4H cows, respectively, that received GnRH and in 22% of P4-saline cows. In Exp. 3, fall-calving multiparous and primiparous cows (d 25 to 31 pp) received either no progestogen (NP; n = 20), P4 via intravaginal devices for 5 d (P4H; n = 21) or melengestrol acetate (MGA; .5 mg.head-1.d-1 for 5 d orally, n = 15). At 48 d after device removal or at 72 h after the last MGA feeding, all cows received 500 micrograms GnRH. Progesterone post-GnRH injection was increased (greater than 1 ng/ml) at d 7 in 64, 100 and 100%, and remained elevated at d 14 in 11, 46 and 100% of NP, MGA and P4H cows, respectively. For all experiments plasma P4 was increased (range 2 to 5 ng/ml) when the devices containing P4 were in place, then decreased (less than 1 ng/ml) by 48 to 50 h after device removal.(ABSTRACT TRUNCATED AT 400 WORDS)     

Influence of nutrition and body condition on pituitary, ovarian, and thyroid function of nonlactating beef cows

Nonpregnant Hereford cows (n = 70) were used to determine the effect of nutrient intake and body condition on reproductive and thyroid function. Body condition scores (BCS; 1 = emaciated; 9 = obese) of cows averaged 5.0 +/- .2 on July 1, and cows were fed for 4 mo either to lose weight and BCS (thin; n = 22), to maintain weight and BCS (moderate; n = 24), or to gain weight and BCS (fat; n = 24). After November 1, cows received a complete ration to maintain weight and BCS. Cows were slaughtered in December (six thin, eight moderate, and eight fat cows) or the subsequent March (16 cows per group). Before slaughter, cows were given two injections of prostaglandin F2 alpha (PGF) 11 d apart. Six days after the second PGF injection, cows were simultaneously treated with 100 micrograms of gonadotropin releasing hormone (GnRH; i.m.) and 100 micrograms of thyrotropin releasing hormone (TRH; i.v.) and serum samples were obtained. The BCS of cows at slaughter (8 d after PGF) averaged 3.4, 5.3, and 7.1 (P less than .01) and carcass energy content averaged 243, 432, and 714 Mcal (P less than .01) for thin, moderate, and fat cows, respectively. Wet ovarian (P less than .001) and corpora lutea (P less than .01) weights were heavier for fat cows. Content of LH in the pituitary gland and concentrations of thyroxine (T4) in serum after GnRH/TRH were not influenced by nutrient intake or BCS. However, thin cows had greater concentrations (P less than .05) of LH in serum after GnRH/TRH than did moderate or fat cows. We conclude that nutrient intake and body energy reserves of beef cows influenced ovarian function and LH in serum after treatment with GnRH.     

Detection of urinary luteinizing hormone in Japanese black cows after administration of gonadotropin-releasing hormone

The blood luteinizing hormone (LH) surge in cows is well studied. However, little is known about urinary LH in cows. This study examined urinary LH concentrations after administration of gonadotropin-releasing hormone (GnRH) in six Japanese black cows to induce LH secretion from the pituitary gland into the bloodstream. Abrupt rises in plasma and urinary LH were observed after GnRH administration. Plasma and urinary LH peaked at 2 and 5 hr, respectively. A positive correlation was observed between plasma LH concentrations and urinary LH amounts. Ovulation was confirmed in the cows after 48 hr of GnRH administration. These data strongly suggest that urinary LH is derived from plasma LH, which triggers ovulation in cows.