Influence of flushing on LH secretion, follicular growth and the response to estrus synchronization treatment in suckled beef cows

The effects of energy supplementation (flushing) on LH and estradiol secretion, follicular growth and the response to estrus synchronization treatment (Norgestomet + PMSG initiated 41.9 +/- 3.4 d after calving) were investigated in 16 suckled beef cows fed either 70% (Group C, n = 8) of energy requirements from calving to 3 wk after AI or fed the same restricted diet until 11 d before synchronization and then were supplemented with 2 kg concentrate until 3 wk after AI (Group S, n = 8). Concentrations of LH and estradiol 17 beta were measured from 3 sampling periods: 25 and 39 d after calving and between 29 and 49 h after implant removal. Ovaries were examined by ultrasonography 11 d before treatment to implant withdrawal (IR). The effects of energy level, day (or hour) of observation and corresponding interactions were tested on repeated measurements by split-plot ANOVA. No positive effect of flushing was observed on characteristics of LH secretion on Day 39. However, the size of the largest follicle and the number of large follicles were higher in Group S than in Group C cows, respectively, 7 and 9 d after the beginning of flushing to 2 d after the start of treatment. After IR, the estradiol secretion tended to be higher in Group S than in Group C cows (9.8 +/- 0.4 pg/mL vs 7.2 +/- 0.2 pg/mL; P = 0.06), but no effect on LH secretion was observed. After implant removal 12 cows ovulated (Group S: 7/8 vs Group C: 5/8; P > 0.05), 7 were pregnant at 21 d after AI (Group S: 6/8 vs Group C: 1/8; P < 0.05) and 4 at 45 d after AI (Group S: 4/8 vs Group C 0/8; P > 0.05). To conclude, flushing had a positive effect on follicular growth, which does not seem to be mediated by LH. In cows fed a restricted diet, flushing enhanced follicular growth, increased the fertilization rate and/or reduced early embryonic death.     

Effects of suckling and of a long interval after ovariectomy on hypothalamo-hypophyseal responsiveness to naloxone, morphine and GnRH in beef cows

The response of serum luteinizing hormone (LH) to morphine, naloxone and gonadotropin-releasing hormone (GnRH) in ovariectomized, suckled (n=4) and nonsuckled (n=3) cows was investigated. Six months after ovariectomy and calf removal, the cows were challenged with 1mg, i.v. naloxone/kg body weight and 1 mg i.v. morphine/kg body weight in a crossover design; blood was collected at 15-minute intervals for 7 hours over a 3-day period. To evaluate LH secretion and pituitary responsiveness, 5 mug of GnRH were administered at Hour 6 on Day 1. On Days 2 and 3, naloxone or morphine was administered at Hour 3, followed by GnRH (5 mug/animal) at Hour 6. Mean preinjection LH concentrations (3.6 +/- 0.2 and 4.7 +/- 0.2 ng/ml), LH pulse frequency (0.6 +/- 0.1 and 0.8 +/- 0.1 pulses/hour) and LH pulse amplitude (2.9 +/- 0.5 and 2.9 +/- 0.6 ng/ml) were similar for suckled and nonsuckled cows, respectively. Morphine decreased (P < 0.01) mean serum LH concentrations (pretreatment 4.2 +/- 0.2 vs post-treatment 2.2 +/- 0.2 ng/ml) in both suckled and nonsuckled cows; however, mean serum LH concentrations remained unchanged after naloxone. Nonsuckled cows had a greater (P < 0.001) LH response to GnRH than did suckled cows (area of response curve: 1004 +/- 92 vs 434 +/- 75 arbitrary units). We suggest that opioid receptors are functionally linked to the GnRH secretory system in suckled and nonsuckled cows that had been ovariectomized for a long period of time. However, gonadotropin secretion appears not to be regulated by opioid mechanisms, and suckling inhibits pituitary responsiveness to GnRH in this model.     

Decreased pulsatile LH secretion in heifers superovulated with eCG or FSH

This study was designed to test the hypothesis that treatment with super-ovulatory drugs suppresses endogenous pulsatile LH secretion. Heifers (n=5/group) were superovulated with eCG (2500 IU) or FSH (equivalent to 400 mg NIH-FSH-P1), starting on Day 10 of the estrous cycle, and were injected with prostaglandin F(2alpha) on Day 12 to induce luteolysis. Control cows were injected only with prostaglandin. Frequent blood samples were taken during luteolysis (6 to 14 h after PG administration) for assay of plasma LH, estradiol, progesterone, testosterone and androstenedione. The LH pulse frequency in eCG-treated cows was significantly lower than that in control cows (2.4 +/- 0.4 & 6.4 +/- 0.4 pulses/8 h, respectively; P<0.05), and plasma progesterone (3.4 +/- 0.4 vs 1.8 +/- 0.1 ng/ml, for treated and control heifers, respectively; P<0.05) and estradiol concentrations (25.9 +/- 4.3 & 4.3 +/- 0.4 pg/ml, for treated and control heifers, respectively; P<0.05) were higher compared with those of the controls. No LH pulses were detected in FSH-treated cows, and mean LH concentrations were significantly lower than those in the controls (0.3 +/- 0.1 & 0.8 +/- 0.1, respectively; P<0.05). This suppression of LH was associated with an increase in estradiol (9.5 +/- 1.4 pg/ml; P<0.05 compared with controls) but not in progesterone concentrations (2.1 +/- 0.2 ng/ml; P>0.05 compared to controls). Both superovulatory protocols increased the ovulation rate (21.6 +/- 3.9 and 23.0 +/- 4.2, for eCG and FSH groups, respectively; P>0.05). These data demonstrate that super-ovulatory treatments decrease LH pulse frequency during the follicular phase of the treatment cycle. This could be explained by increased steroid secretion in the eCG-trated heifers but not in FSH-treated animals.     

The effect of progestin and GnRH treatments on ovarian function and reproductive hormone secretions of anestrous postpartum suckled beef cows

Eighteen anestrous crossbred suckled beef cows were assigned to one of three treatment groups. Treatments were as follows: Group 1 cows (n = 3) were untreated and served as controls, Groups 2 cows (n = 6) were intramuscularly administered 250 mug GnRH, and Group 3 cows (n = 9) were subcutaneously administered a progestin ear implant for eight days prior to the administration of 250 mug GnRH. The GnRH was given to cows in Group 3 24 h after the time of progestin implant removal. Cows were 21 to 31 days postpartum at the time of GnRH treatment. The percent of cows that ovulated after the time of GnRH treatment was 0%, 83% and 100% for Groups 1, 2 and 3, respectively. For the cows that ovulated, more (P < 0.05) cows in Group 2 (80%) had abnormal luteal phases than in Group 3 (33%). The GnRH-induced LH release and peak LH concentrations were greater (P < 0.01) in the cows in Group 3 (214.3 +/- 37.1 ng/ml) than in the cows in Group 2 (142.7 +/- 19.0 ng/ml). The LH concentrations of the control cows remained very low throughout the sampling period. Although prostaglandin metabolite (PGFM) concentrations were not significantly (P > 0.10) different among groups, mean concentrations were higher and more variable for cows in Groups 1 (39.2 +/- 5.2 pg/ml) and 2 (39.4 + 6.1 pg/ml) than for cows in Group 3 (25.1 + 1.4 pg/ml).     

Effects of dietary energy level on luteinizing hormone and adrenal function in the post partum beef cow

The effects of dietary energy and suckling on adrenal function and luteinizing hormone (LH) concentrations were investigated in primiparous postpartum cows. Ten heifers were assigned at calving to either high (22.8 Mcal/day) or low (15.2 Mcal/day) energy diets. Blood samples were collected every 15 minutes for 8 hours on 28, 42, and 56 days post partum. Calves were allowed to suckle ad libitum during sampling periods. Serum samples were analyzed by radioimmunoassay for LH and cortisol. Concentrations of catecholamines were quantified by reverse-phase HPLC. Body weights were decreased (P<0.01) by low energy intake. In addition, low energy diet cows had lower mean LH concentrations (0.97 +/- 0.09 vs 1.57 +/- 0.07 ng/ml), P<0.05) than high energy diet cows. Luteinizing hormone concentrations in high energy diet cows increased with days post partum, resulting in a treatment-by-time interaction (P<0.005). Treatment did not affect mean cortisol concentrations. However, within 15 minutes of suckling cortisol release was significantly above baseline in 77% of the observed suckling events. Dihydroxyphenylalanine (DOPA) increased in high energy diet cows compared with that of low energy diet cows (2,833 +/- 243 vs 1,294 +/- 243 pg/ml, P<0.01). Norepinephrine (NE) and 3,4-dihydroxyphenylacetic acid (DOPAC) were not influenced by treatment. Plasma NE decreased during the postpartum interval (P<0.005). These data suggest that reduced energy intake may prevent the increase in LH associated with increasing days post partum and alter adrenal function. In addition, spontaneous suckling events elicit a release of cortisol.     

Effects of ultrasound-guided transvaginal follicular aspiration on oocyte recovery and hormonal profiles before and after GnRH treatment

Endocrine changes and recovered oocytes were evaluated during 16 wk of ultrasound-guided transvaginal follicular aspiration (TVFA) and prior to and following administration of GnRH at the cessation of aspiration. Nonlactating previously aspirated (PAC, n = 4) and non-aspirated, (AC, n = 4) Holstein cows were subjected to 16 wk of twice-weekly aspiration. Four control cows (OAC) were aspirated 1 time only at the final TVFA session (wk 16). Jugular blood samples were collected from all cows during aspiration, before and after the final TVFA session, and during an 18-d period following cessation of aspiration. Ovarian activity was monitored in all cows after cessation of aspiration for 18 d. The PAC and AC cows averaged 3.4 +/- 1.2 (+/- SE) and 6.8 +/- 1.2 oocytes per session, respectively. Progesterone concentrations during TVFA did not differ between the PAC and AC (0.8 +/- 0.1 and 0.9 +/- 0.1 ng/mL, respectively). Progesterone concentration in OAC was 4.5 +/- 0.2 ng/mL before TVFA, while the PAC and AC averaged 0.5 +/- 0.2 and 0.3 +/- 0.2 ng/mL, respectively, at 16 wk. At Week 16 LH was 1.0 +/- 0.2 ng/mL and it increased to 7.5 +/- 0.1 ng/mL after GnRH treatment. The LH concentration before the final aspiration session was higher at peak amplitude in PAC than in AC groups and peak length was longer in OAC than in AC cows (P < 0.07). Between 18 and 24 h after the last aspiration there were more LH peaks and greater peak frequencies in PAC than in OAC cows (P < 0.07), and the interval between peaks was longer in PAC and AC cows (P < 0.10) than in OAC cows. Mean FSH concentrations were lower (P < 0.01) for OAC than for PAC and AC groups at 20 and 24 h after the last aspiration. Follicle numbers after GnRH varied most among treatment groups for follicles < 9 mm, with the PAC, AC and OAC averaging 5.1 +/- 1.0, 5.1 +/- 1.0, and 3.8 +/- 1.0 follicles/d, respectively. Progesterone concentrations increased to 1.1 +/- 0.3 ng/mL in PAC cows and 2.5 +/- 0.3 and 3.4 +/- 0.3 ng/mL in AC and OAC groups, respectively, during the 18-d period. These results suggest that long-term TVFA affects progesterone, LH and FSH profiles and ovarian dynamics in cows.     

Testosterone inhibits gonadotropin-releasing hormone pulse frequency in the male sheep

The objective was to determine the effect of chronic testosterone (T) treatment on GnRH and LH secretion in wethers. Rams were either castrated only or castrated and immediately treated with Silastic implants containing T. Several weeks later, a device for collecting hypophyseal-portal blood was surgically implanted. Six to seven days later, blood samples were collected simultaneously and continuously from the portal vessels and jugular vein of pairs of conscious animals. Samples were divided at 10-min intervals for 6-12 h. One hour before the end of collection, all animals received i.v. injections of 250 ng of GnRH. In samples collected simultaneously from 6 pairs of animals, T reduced the frequency of both GnRH pulses (1.8 +/- 0.2 vs. 0.9 +/- 0.3/h, p less than 0.03) and LH pulses (1.6 +/- 0.1 vs. 0.8 +/- 0.3/h, p less than 0.03). T did not alter amplitude of either GnRH or LH pulses. Testosterone reduced mean GnRH (9.7 +/- 0.6 vs. 7.9 +/- 0.5 pg/ml, p less than 0.05), whereas mean LH was not significantly reduced (9.6 +/- 1.4 vs. 6.1 +/- 1.8 ng/ml, p = 0.16). These results support the hypothesis that T reduces GnRH pulse frequency.     

The negative feedback effect of estradiol-17beta on secretion of luteinizing hormone in beef cows

Thirty-two ovariectomized cows were used to determine the time course for the negative feedback effect of estradiol-17beta (E) on secretion of the luteinizing hormone (LH). The cows were injected with gonadotropin releasing hormone (GnRH; 40 mug) 2.5 or 5 h after pretreatment with E (1 mug/kg body weight) or with a vehicle for control (C). Pretreatment with E resulted in lower serum concentrations of LH at 2.5 h (0.27 vs 0.90 ng/ml; P < 0.01) and at 5 h (0.27 vs 0.67 ng/ml; P < 0.01); less LH was released in response to GnRH at 2.5 h after treatment compared to cows treated with C (10 +/- 4.9 vs 27 +/- 3.8 ng/ml; P < 0.001). However, when GnRH was administered 5 h after E or C, there was no difference in the total amount of LH released (34 +/- 1.8 vs 26 +/- 4.4 ng/ml; P > 0.2). Time to half area (estimate of decay for the induced surge of LH) was longer for cows treated with E when compared to those treated with C (1.3 vs 0.9 h, P < 0.001; 1.5 vs 0.8 h, P < 0.001). Time to half area was not affected by the time of administration of GnRH after E (P > 0.4). These results suggest that E acts in the pituitary to cause the initial decrease in concentrations of LH. Pituitaries in animals pretreated with E regained the capacity to release as much LH at 5 h after treatment as those treated with C at a time when LH concentrations were still suppressed by E. Thus, the hypothalamus or an extra-hypothalamic area may be involved in maintaining the suppression of LH secretion after the initial effect on the pituitary has declined.     

Effects of season and phase of the estrous cycle on steroidogenesis and LH-FSH sensitivity of large ovine follicles perfused in vitro

The aims of this study were to compare stero?dogenesis (progesterone, androstenedione and estradiol production) and response to LH and FSH challenge by whole perifused follicles 4 to 5.5 mm in diameter, obtained at different periods of the breeding season (onset, middle, end), during anestrus and the luteal phase. We have observed that all follicles do not have the same stero?dogenetic potential and do not respond with the same intensity to LH and FSH. At the middle of the breeding season, LH and FSH supplementation was ineffective in increasing progesterone secretion by follicles (0.19+/-0.05 vs. 0.20+/-0.03 ng/mL). In contrast, gonadotrophin challenge elicited significant (P<0.05) increases in androstenedione (0.94+/-0.34 vs. 0.35+/-0.09 ng/mL) and estradiol (120+/-11 vs. 49+/-10 pg/mL) production immediately after its administration. At the onset of the breeding season, steroidogenesis was identical under both basal and gonadotrophin-stimulated conditions unlike that in middle of the breeding season. However follicles were more sensitive to the gonadotrophin challenge in terms of estradiol production than those collected at the middle of the breeding season (220+/-45 vs. 120+/-11 pg/mL). Follicles obtained at the end of the breeding season featured higher progesterone (2.61+/-0.81 vs. 0.19+/-0.05 ng/mL; P<0.05) and lower estradiol production (10+/-3 vs. 49+/-10 pg/mL; P<0.05) that was not influenced by LH and FSH. Basal androstenedione secretion was comparable to that observed at the middle of the breeding season (0.42+/-0.10 vs. 0.35+/-0.09 ng/mL), but the response to stimulation was significantly higher (1.82+/-0.61 vs. 0.94+/-0.34 ng/mL; P<0.05). In anoestrus and the luteal phase, follicles presented higher progesterone and androstenedione and lower estradiol concentrations (P<0.05) compared with those obtained during the follicular phase at the middle of the breeding season. In the luteal phase, follicles remained capable of responding to LH-FSH challenge by increasing estradiol secretion (9+/-1 before and 21+/-6 pg/mL after LH-FSH; P<0.05). In contrast, in the luteal phase, estradiol production was not increased by LH-FSH challenge (7+/-2 vs. 12+/-4 pg/mL).     

Failure of naloxone to stimulate luteinizing hormone secretion during pregnancy and steroid treatment of ovariectomized beef cows

The response of serum luteinizing hormone (LH) to naloxone, an opiate antagonist, and gonadotropin-releasing hormone (GnRH) was measured in cows in late pregnancy to assess opioid inhibition of LH. Blood samples were collected at 15-min intervals for 7 h. In a Latin Square arrangement, each cow (n = 6) received naloxone (0, 0.5, and 1.0 mg/kg BW, i.v.; 2 cows each) at Hour 2 on 3 consecutive days (9 +/- 2 days prepartum). GnRH (7 ng/kg body weight, i.v.) was administered at Hour 5 to all cows on each day. Mean serum LH concentrations (x +/- SE) before naloxone injection were similar (0.4 +/- 0.1 ng/ml), with no serum LH pulses observed during the experiment. Mean serum LH concentrations post-naloxone were similar (0.4 +/- 0.1 ng/ml) to concentrations pre-naloxone. Mean serum LH concentrations increased (p less than 0.05) following GnRH administration (7 ng/kg) and did not differ among cows receiving different dosages of naloxone (0 mg/kg, 1.44 +/- 0.20; 0.5 mg/kg, 1.0 +/- 0.1; 1.0 mg/kg, 0.9 +/- 0.1 ng/ml). In Experiment 2, LH response to naloxone and GnRH was measured in 12 ovariectomized cows on Day 19 of estrogen and progesterone treatment (5 micrograms/kg BW estrogen: 0.2 mg/kg BW progesterone) and on Days 7 and 14 after steroid treatment. On Day 19, naloxone failed to increase serum LH concentrations (Pre: 0.4 +/- 0.1; Post: 0.4 +/- 0.1 ng/ml) after 0, 0.5, or 1.0 mg/kg BW.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of the effects of two GnRH antagonists on LH and FSH secretion,follicular growth and ovulation in the mare

The effects of two GnRH antagonists were tested in order to delay and/or synchronise ovulation in mares. Five mares received Antarelix (0.01 mg.kg(-1)), 5 mares received Cetrorelix (the same dose), 5 mares (control mares) received the vehicle intravenously, twice daily, for 8 days from the day the largest follicle reached 22 mm following prostaglandin administration. Ovulation was postponed in all mares injected with Antarelix (19.4 +/- 1.2 days after the beginning of the treatment) and in 2/5 mares injected with Cetrorelix (20 +/- 1 days) vs. 6.2 +/- 0.4 days in control mares. During the treatment, LH concentrations were strongly depressed in Antarelix and in Cetrorelix mares (1.6 +/- 0.1 and 3.8 +/- 0.5 ng.mL(-1) respectively vs. 21 +/- 2.5 ng.mL(-1) in control mares). In the 3 Cetrorelix mares which ovulated during the treatment. 2 initiated their LH surge at this moment. FSH concentrations were not affected in Antarelix or in Cetrorelix mares during the treatment (11.4 +/- 1.3 and 7.9 +/- 0.8 ng.mL(-1) respectively vs. 10.5 +/- 0.8 ng.mL(-1) in control mares). In conclusion, Antarelix seems more efficient than Cetrorelix for postponing ovulation in mares. The role of LH in antral follicular development before the preovulatory stage is confirmed.     

Effect of a long-lasting opioid receptor antagonist (naltrexone) on pulsatile LH release in early postpartum Holstein dairy cows

The present trial was the first one to investigate the effect of an intravenous injection of naltrexone, an opioid receptor antagonist that has a longer duration of action than that of naloxone, on the LH pulse in early postpartum Holstein dairy cows. On Day 10 postpartum, blood samples were collected from cows at 10-min intervals for a period of 4 h before (pre-injection period) and a period of 5 h after (post-injection period) an intravenous injection of 10 mL of saline (Control Group, n=5) or 300 mg of naltrexone in 10 mL of saline (Naltrexone Group, n=5). The plasma LH level was assayed by double antibody radioimmunoassay. The number of LH peaks per 1 h, the mean LH level, and the amplitude of LH peaks were analyzed utilizing the Pulsar algorithm, and data were compared by repeated measures ANOVA. No differences were observed in the parameters of LH pulse in the pre-injection period between the Control and the Naltrexone Groups (P>0.10). In the Naltrexone Group, the number of LH peaks per 1 h and the mean LH level were significantly higher in the post-injection period than in the pre-injection period (0.85 +/- 0.29 vs. 1.24 +/- 0.17, P<0.05, and 1.81 +/- 0.70 vs. 2.47 +/- 0.92 ng/ml, P<0.05, respectively), but there was no significant increase in the amplitude of LH peaks (1.48 +/- 0.64 vs. 1.83 +/- 0.82 ng/ml, P>0.10). In contrast, all of the parameters of LH pulse remained unchanged in the Control Group (P>0.10). These results suggested that an intravenous injection of naltrexone activates the LH pulse.     

Effect of nutrition on the LH response to calf removal and GnRH

Ten primiparous crossbred cows were assigned to two dietary groups at calving. One group received 120% and the other group received 80% of the National Research Council (NRC) recommended allowance of dietary energy for primiparous cows. At 60 days postpartum, calves were removed from their dams. Blood samples were collected from the cows at 15-min intervals for 8 hr beginning at the time of calf removal and again 24 hr, 48 hr and 72 hr after calf removal. At 72 hr after calf removal, all cows were given 200 ug GnRH intravenously. At calf removal, serum LH concentrations were higher (P<0.01) for cows on 120% (0.9 +/- 0.03 ng/ml) compared to cows on 80% (0.5 +/- 0.03 ng/ml) of recommendations. Serum LH concentrations increased (1.6 +/- 0.1 ng/ml, P<0.01) by 24 hr in cows on the highenergy diet. In contrast, a similar increase was not observed in cows on the low-energy diet until 48 hr after calf removal (1.4 +/- 0.2 ng/ml, P<0.01). These contrasting patterns in serum LH concentrations resulted in a diet by time interaction (P<0.01). Serum LH concentrations increased in both dietary energy groups following GnRH injection, but the response was greater (P<0.01) in cows on the low-energy diet compared to the cows fed the high-energy diet. These results indicate that inadequate dietary energy delays the LH response to calf removal and increases the LH response to exogenous GnRH.     

Oestradiol-17beta responsiveness, plasma LH profiles, pituitary LH and FSH concentrations in long-term ovariectomised Holstein cows at 24 h, 48 h and 21 days following treatment with an absorbable GnRH agonist implant

Non-lactating OVX Holstein cows (N = 34) were used to investigate the effect of s.c. placement of an absorbable GnRH agonist implant (Ovuplant; deslorelin 2.1mg, Peptech Animal Health, Australia) on the relationship of plasma LH, oestradiol responsiveness and pituitary LH content. On the day of implant insertion (Day 0), one group (OVU-48h; N = 5) received Ovuplant and had blood samples collected at hourly intervals to characterize the LH response, while a second group (CON-48 h; N = 5) remained untreated and acted as controls. Blood samples were collected every 10 min over 6 h from CON-48 h and OVU-48 h, at 24 h post-implant insertion. These cows were then slaughtered at 48 h post-implant insertion and their pituitaries recovered. Another group received Ovuplant (OVU-21d+E2; N = 10) or were left untreated (CON-21d+E2) and 21 days later were injected i.m. with 0.5 mg 17beta-E2. Blood samples were collected every 10 min for 4 h on the day before E2 injection to characterize LH pulse frequency and amplitude. Beginning 14 h later, blood samples were collected hourly for 12 h to characterize the expected LH surge. These cows were slaughtered and their pituitary glands recovered and assayed for LH and FSH content. Peak plasma LH concentrations (59 +/- 19 ng/ml) were measured after 30 min of Ovuplant insertion. They had returned to pre-treatment levels by 7 h. By 24 h post-implant insertion, OVU-48 h plasma LH profiles were characterized by reduced LH pulse frequency (0.23 +/- 0.09 pulses/h versus 0.75 +/- 0.26 pulses/h; OVU-48 h versus CON-48 h; P < 0.05). The cows that received Ovuplant had lower LH pulse amplitude, LH pulse frequency and mean LH concentrations after 20 days. Injection of 0.5 mg 17beta-E2 induced an LH surge in every one of the control cows with their peak concentrations measured 18 h post injection. No increase in LH was detected in any Ovuplant treated cows. Pituitary FSH content was reduced in Ovuplant treated cows after 48 h, but not that of LH. In conclusion, absorbable deslorelin implants induced a substantial but temporary release of LH, but even 21 days later their LH profiles were characterized by marked suppression of pulsatile LH and an absence of response to E2. These results suggest the implant has prolonged biological activity.     

Resumption of follicular activity in the early post-partum period of dairy cows

Lactating Friesian dairy cows (2nd-4th parity) which calved in spring (N = 7) or autumn (N = 15) were used. Their ovaries were examined by ultrasound scanning and blood samples were obtained daily for progesterone and oestradiol concentrations from the 5th day after calving until the first post-partum ovulation occurred. Five autumn-calving cows selected at random were bled every 15 min over a 6-h period on 1 day each week for 4 weeks after calving to assess the patterns of LH secretion. Follicular development during the post-partum anoestrous period was characterized by the growth and regression of small (less than or equal to 4 mm) and medium-sized (5-9 mm) follicles, until a dominant follicle (greater than 10 mm) was detected. The first detected dominant follicle ovulated in 14 cows, became cystic in 4 cows (all in autumn), and failed to ovulate in 1 cow. It was not possible to detect a dominant follicle in 3 cows due to scanning difficulties. The post-partum interval to detection of the first dominant follicle (mean +/- s.d.) was shorter (P less than 0.05) in autumn (6.8 +/- 1.8 days) than in spring (20 +/- 10.1 days). However, there was no significant difference between the respective intervals to first ovulation (autumn 27.4 +/- 25.9 and spring 27.3 +/- 18.9 days). Autumn-calved cows which had cysts had longer (P less than 0.001) intervals to first ovulation (58.2 +/- 23.5 days) than did normal cows (12.0 +/- 2.5 days). All cows with cysts had twin ovulations at their first post-partum ovulation. A pulsatile pattern of LH secretion was detected in the first week post-partum and LH pulse frequency was 2-3 per 6-h period in Weeks 1 and 2 post partum and increased to 5-7 pulses per 6-h period in the presence of a dominant or cystic follicle. Concentrations of progesterone in plasma during post-partum anoestrus were usually low (less than 0.2 ng/ml); oestradiol concentrations were also low (less than 5 pg/ml), but higher values (5-110 pg/ml) were observed in cows that had a dominant or a cystic follicle.     

Relationship between serum luteinizing hormone and estradiol in prepubertal boars

Effects of estradiol on serum luteinizing hormone (LH) were studied in prepubertal boars. In Exp. 1, 15-wk-old boars were given (iv) 50 mug estradiol, 1 mg testosterone or 1.5 ml ethanol. Estradiol (P<0.05) decreased LH over a 2.5-hr period, but testosterone did not. In Exp. 2, an estradiol implant reduced LH sample variance (P<0.01) while LH (547 +/- 96 vs 655 +/- 43 pg/ml) and estradiol (14.2 +/- 3.3 vs 18.4 +/- 1.0 pg/ml; control vs implant) were unchanged in 12-wk-old boars. Pulsatile LH releases (4.3 +/- 1.1 vs 3.0 +/- 0.4 pulses/pig/8 hr; control vs treated) and pulse amplitude (272 +/- 34 vs 305 +/- 40 pg/ml) were not affected. The implant tended to decrease serum testosterone (4.86 +/- 0.75 vs 7.66 +/- 1.51 ng/ml; P<0.10). In Exp. 3, LH was higher after zero implants than after four implants (279 +/- 7 vs 227 +/- 9 pg/ml; P<0.01), and LH after two implants was also higher than after four implants (263 +/- 7 pg/ml; P<0.01) in 14-wk-old boars in a Latin square design. Peak LH after 40 mug gonadotropin releasing hormone (GnRH) was less after two and four implants (1,100 +/- 126 and 960 +/- 167 pg/ml, respectively; P<0.01) than after zero implants (1,742 +/- 126 pg/ml). Slope of the first 20 min of LH response to GnRH was greater after zero implants (45.3 pg/min; P<0.05) than after either two or four implants (20.6 and 16.9 pg/min, respectively). Implant treatment decreased serum testosterone (P<0.025) but increased estradiol (P<0.10). Small changes in serum estradiol resulted in changes in LH. These changes in sample variance and mean LH were recognized by boars as different from normal because serum testosterone decreased. Changes in LH may result from estradiol's negative effect on pituitary responsiveness to endogenous GnRH because response to exogenous GnRH was depressed by estradiol.     

Estradiol and testosterone concentrations increase with fasting in weaned pups of the northern elephant seal (Mirounga angustirostris)

Although neonatal development is generally associated with increased levels of circulating testosterone (T) and estradiol (E2), food deprivation may inhibit steroidogenesis. Therefore, these potentially conflicting stimuli were examined in fasting weaned northern elephant seal (Mirounga angustirostris) pups by measuring serum concentrations of T, E2, progesterone (P4), and luteinizing hormone (LH) by either radioimmunoassay (P4, LH) or enzymeimmunoassay (T, E2). Blood samples were obtained from 20 male and 20 female pups at both early (<1 wk postweaning) and late (6-8 wk postweaning) periods during their natural postweaning fast. T in males (early: 2.9 +/- 0.4 ng/mL; late: 16 +/- 2 ng/mL; P < 0.0001) and E2 in females (early: 42 +/- 6 pg/mL; late: 67 +/- 5 pg/mL; P < 0.01) increased between the two measurement periods, while P4 (early: 2.5 +/- 0.3 ng/mL; late: 2.1 +/- 0.3 ng/mL; P > 0.05) did not. LH increased (early: 46 +/- 4 pg/mL; late: 65 +/- 6 pg/mL; P < 0.05) in males but not in females (early: 69 +/- 9 pg/mL; late: 65 +/- 6 pg/mL; P > 0.05). Increases in LH and T suggest that LH may stimulate T secretion. Alternatively, relatively low concentrations of LH in both males and females may reflect negative feedback inhibition imposed by elevated T and E2 concentrations. Despite the inherent postweaning fast, concentrations of T and E2 increased, suggesting that they may be critical for the continued development of pups. Therefore, compensatory mechanisms may exist that alleviate the fasting-induced inhibition of gonadal steroidogenesis during neonatal development in elephant seal pups.     

Relationship between peripheral estrogen concentrations at insemination and subsequent fetal loss in cattle

In a survey on pregnancy rate and embryonic losses in dairy cattle on 6 Israeli farms, cows (n = 78) were divided into 3 groups on the basis of ultrasonography at 21 d post insemination; pregnancy diagnosis at 40 to 50 d post insemination and blood progesterone (P4) levels at 21 d. The groups were either pregnant (P4 level > 1.0 ng/ mL); not pregnant (P4 < 0.5 ng/mL), or showed early embryo loss (P4 > 1.0 ng/mL and the presence of an embryonic vesicle on D 21 but later returned to estrus or were found not pregnant on D 40 to 50). On the day of insemination, peripheral estrogen was significantly higher (P < 0.05) in the early embryo loss group (15.3 +/- 1.1 pg/mL, n = 27) than in pregnant (9.4 +/- 0.6 pg/mL, n = 26) or not pregnant (9.6 +/- 0.7 pg/mL, n = 25) group. The cows on 3 farms which were fed 1 to 2 kg/d of vetch (Vicia sativa), an estrogenic legume, had higher estrogen concentrations on the day of insemination than cows (2 farms) fed other legumes (13.7 +/- 0.64, n = 58 vs 10.7 +/- 0.8 pg/mL, n = 42; P < 0.01). On one of the 3 farms, vetch was replaced with alfalfa after the first year. Following the cessation of vetch feeding the estrogen concentrations in the blood decreased from 32 +/- 5 pg/mL to 14 +/- 2 pg/mL (n = 9). These data suggest that high peripheral estrogen on the day of insemination is associated with early embryonic loss. These data also indicate that estrogen concentrations on the day of insemination can be influenced by diet.     

Restoration of LH output and 17beta-oestradiol responsiveness in acutely ovariectomised holstein dairy cows pre-treated with a GnRH agonist (deslorelin) for 10 days

The objectives of the study were firstly to identify the role of the ovary in maintaining plasma luteinising hormone (LH) concentrations in cows treated with an implant of a potent GnRH agonist (deslorelin), and secondly to characterise the changes in LH following ovariectomy (OVX) in the same animals. Oestrus was synchronised in mature Holstein dairy cows and deslorelin implants were inserted 17 days later into two-third of the cows. A further 10 days later (day 0) all cows had bilateral OVX performed. A control group (CON; n=4) received no treatment and had blood samples collected at 15-min intervals for 8h on the day prior to OVX (day -1) and similarly on days 4 and 10. One group (DES_IN; n=4) had implants in place for the duration of the study while another group had implants removed (DES_OUT; n=4) at the time of OVX. DES_IN cows were sampled hourly at each sampling session (days -1, +4 and +10), whereas DES_OUT cows were sampled similarly to CON except on day -1 when hourly samples were collected.Predictable post-operative increases in mean LH (0.61 ng/ml versus 1.79 ng/ml; P<0.01) and LH pulse amplitude (0.66 ng/ml versus 1.56 ng/ml; day -1 versus day +10; P<0.01) occurred after CON cows were ovariectomised. Smoothed LH means showed a delayed effect of time compared to arithmetic means. Pulse frequency was unchanged following OVX in CON cows. A comparison of all cows that had been treated with deslorelin from day -1 showed a significant elevation of smoothed mean LH compared to untreated cows (0.80 ng/ml versus 0.34 ng/ml; DES_IN and DES_OUT versus CON; P<0.05). DES_IN cows had a 54% reduction in mean LH from day -1 to +4 following OVX (1.05 ng/ml versus 0.48 ng/ml; P<0.01) indicating the probable involvement of the ovary in the maintenance of elevated basal LH. No further reduction was detected by day +10. The LH response to an intramuscular (IM) injection of 500 microg 17beta-oestradiol (E2) on day +11 varied significantly between treatment groups (P<0.01). CON cows showed a typical LH surge, reaching maximum concentrations (10.3 ng/ml) at 17.3h post-injection. Even though low amplitude LH pulsatility had been restored in DES_OUT cows by day +4, there was an inconsistent response to E2 on day +12; one cow had an apparently normal surge yet, others showed only attenuated responses. Pulse amplitude in DES_OUT cows was lower at days +4 and +10 compared to CON (P<0.05). DES_IN cows did not produce any surge after E2. Mean LH prior to OVX (day -1) remained unchanged following the 500 microg oestradiol injection (0.38 ng/ml versus 0.45 ng/ml pre-E2 versus post-E2 compared to 1.05 ng/ml pre-OVX).The results of this experiment implicated ovarian involvement in maintaining elevated basal LH output in cows that were chronically treated with a GnRH agonist. Individual cows varied in their LH surge response to exogenous E2 given 12 days after implant removal, even though LH pulse amplitude and frequency had been restored.