Induction of ovulation in the acyclic postpartum ewe following continuous, low-dose subcutaneous infusion of GnRH

Pituitary and ovarian responses to subcutaneous infusion of GnRH were investigated in acyclic, lactating Mule ewes during the breeding season. Thirty postpartum ewes were split into 3 equal groups; Group G received GnRH (250 ng/h) for 96 h; Group P + G was primed with progestagen for 10 d then received GnRH (250 ng/h) for 96 h; and Group P received progestagen priming and saline vehicle only. The infusions were delivered via osmotic minipumps inserted 26.6 +/- 0.45 d post partum (Day 0 of the study). Blood samples were collected for LH analysis every 15 min from 12 h before until 8 h after minipump insertion, then every 2 h for a further 112 h. Daily blood samples were collected for progesterone analysis on Days 1 to 10 following minipump insertion, then every third day for a further 25 d. In addition, the reproductive tract was examined by laparoscopy on Day -5 and Day +7 and estrous behavior was monitored between Day -4 and Day +7. Progestagen priming suppressed (P < 0.05) plasma LH levels (0.27 +/- 0.03 vs 0.46 +/- 0.06 ng/ml) during the preinfusion period, but the GnRH-induced LH release was similar for Group G and Group P + G. The LH surge began significantly (P < 0.05) earlier (32.0 +/- 3.0 vs 56.3 +/- 4.1 h) and was of greater magnitude (32.15 +/- 3.56 vs 18.84 +/- 4.13 ng/ml) in the unprimed than the primed ewes. None of the ewes infused with saline produced a preovulatory LH surge. The GnRH infusion induced ovulation in 10/10 unprimed and 7/9 progestagen-primed ewes, with no significant difference in ovulation rate (1.78 +/- 0.15 and 1.33 +/- 0.21, respectively). Ovulation was followed by normal luteal function in 4/10 Group-G ewes, while the remaining 6 ewes had short luteal phases. In contrast, each of the 7 Group-P + G ewes that ovulated secreted progesterone for at least 10 d, although elevated plasma progesterone levels were maintained in 3/7 unmated ewes for >35 d. Throughout the study only 2 ewes (both from Group P + G) displayed estrus. These data demonstrate that although a low dose, continuous infusion of GnRH can increase tonic LH concentrations sufficient to promote a preovulatory LH surge and induce ovulation, behavioral estrus and normal luteal function do not consistently follow ovulation in the progestagen-primed, postpartum ewe.     

Effect of ram exposure at the end of progestagen treatment on estrus synchronisation and fertility during the breeding season in ewes

Two experiments were conducted to examine the effects of ram exposure during the breeding season, in combination with progestagen treatment on estrus synchronization, fertility the LH surge and ovulation in ewes. Experiment 1 was subdivided into experiments 1a and 1b. In all experiments cross-bred ewes were treated with an intravaginal sponge for 12-14 days and three days before sponge withdrawal ewes were divided into control (no further treatment; n=191, 103 and 50 for experiments 1a, 1b and 2, respectively) or ram exposed (three mature rams per 50 ewes were introduced; +Ram; n=187, 99 and 49 for experiments 1a, 1b and 2, respectively). At sponge withdrawal ewes in Experiments 1a and 2 received 500 IU eCG and rams were removed from all the +Ram groups. In Experiments 1a and 1b, raddled, entire rams were introduced to ewes 48 h after sponge withdrawal. The timing of mating was recorded and ewes were maintained until lambing. In Experiment 2, estrus behavior was determined every 4 h and the time of the LH surge and ovulation were determined from a subset of 10 ewes per group. In Experiment 1a, less +Ram ewes were bred by 48 h after ram introduction (control 98% versus +Ram 89%, P<0.001) and in Experiments 1a and 1b 14% fewer (P<0.05) of the ewes bred in the first 3 h after ram introduction lambed to that service. In Experiment 1a, ram exposed ewes had a lower litter size than control ewes (1.93+/-0.06 versus 1.70+/-0.06 lambs per ewe; P<0.05). In Experiment 2, rams advanced (P<0.05) estrus, the LH surge and ovulation by 2-6 h compared with control ewes. We speculate that exposure of ewes to rams increased LH secretion and that this in turn increased follicle development and the production of oestradiol that led to a more rapid onset of estrus, the LH surge and ovulation compared to control ewes. Unexpectedly, ewes that were bred had lower fertility in the +Ram groups than control groups.     

Inter-breed variation in the postpartum ewe response to continuous low dose infusion of GnRH

During the nonbreeding season the pituitary and ovarian responses to a subcutaneous GnRH infusion were investigated in acyclic, lactating Mule ewes which exhibit a deep seasonal anestrus and in Finn x Dorset ewes in which seasonal anestrus is ill-defined. Each breed received 10 d of progestagen priming before being subdivided into 3 groups. In Group L + G, 5 lactating ewes received GnRH (250 ng/h sc) for 96 h; in Group D + G, 5 dry ewes received GnRH (250 ng/h sc) for 96 h; in Group L, 5 lactating ewes received saline vehicle for 96 h. The infusions began when lactating and dry ewes were approximately 28 d and 120 d post partum, respectively. Blood samples were collected for LH, progesterone and estradiol analysis. Estrous behavior was monitored between Day -4 and Day +7. On Day +7 the reproductive tract was also examined. In the Mule ewes the mean plasma LH concentration increased (P < 0.05) following minipump insertion in each treatment group, although mean LH levels were greater (P < 0.05) in Group D + G, than in either Group L + G or Group L. Following the GnRH infusion, mean plasma estradiol levels increased (P < 0.05) in Group D + G but not in Group L + G. A preovulatory LH surge and subsequent ovulation occurred in 5 5 , 2 5 and 0 5 ewes from Group D + G, L + G and L, respectively, and estrus was recorded in 5 5 , 1 5 and 0 5 of these ewes, respectively. The LH surges began earlier (P < 0.05) (43.2 +/- 6.8 h vs 77.0 +/- 1.0 h) and the ovulation rate was greater (2.2 +/- 0.37 vs 1.00 +/- 0.00) in Group D + G than Group L + G. In the Finn x Dorset ewes mean LH concentrations increased (P < 0.05), to a similar level following minipump insertion in Groups D + G and L + G, but not Group L. The elevated LH levels were accompanied by increased (P < 0.05) plasma estradiol levels in Group D + G, but not in Group L + G. The GnRH infusion culminated in an LH surge and estrous behavior in 5 5 , 1 5 and 0 5 ewes from Groups D + G, L + D and L, respectively. The interval to the LH surge was similar between Group D + G (48.4 +/- 6.6 h) and Group L + G (46.0 h). Ovulation was evident in those ewes which exhibited an LH surge plus one additional ewe from Group L + G. The mean ovulation rate was greater in Group D + G (4.00 +/- 1.05) than in Group L + G (1.5 +/- 0.50). These data show that continuous GnRH infusion can consistently induce out of season breeding in the nonlactating Mule and Finn x Dorset ewe but can not break combined seasonal and lactational anestrous in these breeds. Further, between-breed differences are evident in the site along the hypothalamic-pituitary-ovarian axis at which reproduction is compromised in ewes at the same chronological stage post partum.     

Stag exposure advances the LH surge and behavioral estrus in Eld's deer hinds after CIDR device synchronization of estrus

The impact of male presence or absence on the timing of the preovulatory LH surge and estrus was studied in 3 experimental groups (n = 6/group) of Eld's deer hinds pretreated with intravaginal progesterone-releasing devices (CIDR-type G) as follows: Group 1 = indirect male contact barn; Group 2 = direct male contact barn; and Group 3 = male isolation barn. For all hinds, the duration of the preovulatory LH surge averaged 2.5+/-0.5 h, whereas mean peak preovulatory and basal LH concentrations were 2.9+/-0.2 ng mL(-1) and 0.27+/-0.03 ng mL(-1), respectively. Nine of 12 male-exposed hinds exhibited a preovulatory LH surge within 24 to 32 h postCIDR device withdrawal, whereas 0 of 6 male-isolated hinds exhibited a preovulatory LH surge during the same time period. Onset of behavioral estrus (45.2+/-2.3, 52.7+/-5.7 and 66.3+/-1.8 h, respectively) was significantly advanced (P<0.05) after CIDR device withdrawal in male exposed hinds (Groups 1 and 2) compared with male isolated hinds (Group 3). These data suggest that stag exposure is important for modulating the timing of the preovulatory LH surge and behavioral estrus after synchronization of estrus with exogenous progestagens.     

Effect of adrenocorticotrophic hormone (ACTH1-24) on ovine pituitary gland responsiveness to exogenous pulsatile GnRH and oestradiol-induced LH release in vivo.

The present experiment was designed to determine if and how exogenous ACTH replicates the effects of stressors to delay the preovulatory LH surge in sheep. Twenty-four hours after oestrous synchronisation with prostaglandin in the breeding season, groups of 8-9 intact ewes were injected with 50 microg oestradiol benzoate (0 h) followed 8 h later by 3 injections of saline or GnRH (500 ng each, i.v.) at 2 h intervals (controls). Two further groups received an additional 'late' injection of ACTH (0.8 mg i.m.) 7.5 h after oestradiol, i.e., 0.5 h before the first saline or GnRH challenge. To examine if the duration of prior exposure to ACTH was important, another group of ewes was given ACTH 'early', i.e. 2.5 h before the first GnRH injection. The first GnRH injection produced a maximum LH response of 1.9+/-0.4 ng/ml which was significantly (p < 0.01) enhanced after the second and third GnRH challenge (7.1+/-1.5 ng/ml and 7.0+/-1.7 ng/ml, respectively; 'self-priming'). Late ACTH did not affect the LH response after the first GnRH challenge (1.9+/-0.4 vs. 1.8+/-0.3 ng/ml; p > 0.05) but decreased maximum LH concentrations after the second GnRH to 35% (7.1+/-1.5 vs. 4.6+/-1.1 ng/ml; p = 0.07) and to 40% after the third GnRH (7.0+/-1.7 vs. 4.0+/-0.8 ng/ml; p = 0.05). When ACTH was given early, 4.5 h before the second GnRH, there was no effect on this LH response suggesting that the effect decreases with time after ACTH administration. Concerning the oestradiol-induced LH surge, exogenous GnRH alone delayed the onset time (20.5+/-2.0 vs. 27.8+/-2.1 h; p > 0.05) and reduced the duration of the surge (8.5+/-0.9 vs. 6.7+/-0.6 h; p > 0.05). The onset of the LH surge was observed within 40 h after oestradiol on 29 out of 34 occasions in the saline +/- GnRH treated ewes compared to 11 out of 34 occasions (p < 0.05) when ACTH was also given, either late or early. In those ewes that did not have an LH surge by the end of sampling, plasma progesterone concentrations during the following oestrous cycle increased 2 days later suggesting a delay, not a complete blockade of the LH surge. In conclusion, we have revealed for the first time that ACTH reduces the GnRH self-priming effect in vivo and delays the LH surge, at least partially by direct effects at the pituitary gland.     

Estrus synchronization and artificial insemination of hair sheep ewes in the tropics

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2alpha (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2alpha injection or at CIDR or sponge removal, sterile rams were placed with the ewes. Jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at -20 degrees C until LH, and progesterone concentrations were determined by RIA. There was no difference (P>0.10) in time to estrus among the CIDR-, PGF2alpha- or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2alpha-treated ewes showed signs of estrus by this time (P<0.06). The time from ram introduction to ovulation was not different (P>0.10) among the CIDR-, PGF2alpha- or sponge-treated ewes. The time to the preovulatory LH surge was similar (P>0.10) among CIDR, PGF2alpha and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P>0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2alpha and bred by TAI using frozen-thawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9% (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2alpha. Even though there were no significant differences in the timing of ovulation or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technician.     

A new strategy for superior reproductive performance of ewes bred out-of-season utilizing progestagen supplement prior to withdrawal of intravaginal pessaries

Two experiments were conducted to examine the effect of progestagen supplement 24h prior to intravaginal pessary withdrawal on reproductive performance of seasonal anestrous ewes. Ewes in each experiment were allocated to treatment and control and all were induced to estrus using either intravaginal MAP (Exp. 1; n=24) or CIDR-G (Exp. 2; n=28) pessaries for 12 days. Half of the ewes in each experiment were supplemented 24h before withdrawal of pessaries with either 10mg oral MAP tablets (Exp. 1) or 25mg i.m. progesterone (P(4)) administration (Exp. 2; P(4)-supplement-treated group). Fertile rams were allowed with the ewes at sponge removal (Day 0, 0h) and estrus was monitored at 6-h intervals for 3 days. Blood samples were collected for measurements of P(4) (Exp. 1 and Exp. 2) and LH (Exp. 2). In both experiments, the percent of ewes in estrus was greater (P<0.05) and intervals to estrus were longer (P<0.05) in progestagen-supplement-treated than control ewes. In Exp. 2, the occurrence and magnitude of LH surges were greater (P<0.01) and intervals to onset of LH surge were longer (P<0.01) in P(4)-supplement-treated than control ewes. In Exp. 2, P(4) supplement elevated P(4) levels from 1.8+/-0.1ng/mL on Day -1 to 4.2+/-0.3 on Day 0 (P<0.001). Following pessaries removal, P(4) concentrations fell to basal values on Day 1 in both groups and remained low until Day 5. Then, P(4) concentrations increased and remained elevated through Day 19 in all (100%) progestagen-supplement-treated in Exp. 1 (12/12) and Exp. 2 (14/14) and in only 5/12 (41.7%) and 6/14 (42.9%) control ewes, respectively. These ewes were confirmed pregnant by ultrasonography and lambed on Day 149.2+/-0.2 following Day 0. In conclusion, progestagen supplement 24h prior to removal of pessary can be used successfully to improve reproductive performance of ewes bred out-of-season.     

Effect of estrus synchronization with Syncro-Mate-B((R)) on serum luteinizing hormone, progesterone and conception rate in Brahman heifers

Twenty-two estrous cyclic, 2-yr-old Brahman heifers were randomly assigned to receive either estrus synchronization with Syncro-Mate-B((R)) (SMB; 11) or no treatment (Control; 11). Blood samples were collected via tail vessel puncture at onset of estrus and daily thereafter until Day 11 after estrus. Blood samples were also collected from five SMB and five Control heifers at 0, 4, 8 and 12 h after the onset of estrus. All samples were processed to yield serum and stored at -20 degrees C until radioimmunoassay. Heifers were inseminated by one technician using semen from a single ejaculate of a Brahman bull 12 h after the onset of estrus. All SMB heifers exhibited estrus within 72 h of implant removal. All heifers had corpora lutea (CL) detected by rectal examination 8 to 12 d following estrus. Serum luteinizing hormone (LH) was not affected by treatment, time (4 - h intervals) or an interaction of treatment by time (P > 0.10). Independent analysis with h indicated that at h 12, SMB (2.2 +/- 0.06 ng/ml) had lower LH than did control heifers (8.9 +/- 2.1 ng/ml). Serum progesterone increased from Day 1 through Day 12 in all heifers, which is indicative of functional CL. Serum progesterone was affected by treatment (P < 0.0001) and time (d intervals; P < 0.10). Progesterone elevation was lower (P < 0.05) and area under the progesterone curve was lower (P < 0.03) in SMB (5.6 +/- 0.5 ng/ml, 32.0 +/- 4.5 units, respectively) when compared with control heifers (7.0 +/- 4 ng/ml, 43.7 +/- 2.4 units, respectively). Conception rate was lower (P < 0.01) in SMB heifers (2 of 11) than in control heifers (8 of 11). The lowered conception rate in SMB treated Brahman heifers may be due to altered timing of LH release following estrus, resulting in an altered time of ovulation.     

How the FSH/LH ratio and dose numbers in the p-FSH administration treatment regimen, and insemination schedule affect superovulatory response in ewes

We wished to evaluate the effects of FSH/LH ratio and number of doses of p-FSH during a superovulatory treatment on ovulation rate and embryo production (Experiment I). In Experiment II, we studied the efficacy of fertilization after various insemination schedules in superovulated donors. In Experiment I estrus was synchronized in 40 ewes (FGA, for 9 days plus PGF2alpha on Day 7) and the ewes were randomly assigned to four treatment groups as follows (n = 10 ewes each): Group A: four p-FSH doses with the FSH/LH ratio held constant (1.6); Group B: four p-FSH doses with the FSH/LH ratio decreasing (FSH/LH 1.6-1.0-0.6-0.3); Group C: eight p-FSH doses with the FSH/LH ratio held constant (1.6); Group D: eight p-FSH doses and FSH/LH ratio decreasing (1.6-1.6, 1.0-1.0, 0.6-0.6, 0.3-0.3). p-FSH administrations were performed twice daily 12 h apart. The ewes were mated at the onset of estrus and again after 12 and 24 h; then, one ram per four ewes was maintained with the ewes for two additional days. Ovarian response and embryo production were assessed on Day 7 after estrus. Experiment II. Three groups (n = 10 each) of superovulated ewes were inseminated as follows: Group M: mated at onset of estrus; Group AI: artificial insemination 30 h after onset of estrus; M + AI) mating at onset of estrus and intrauterine AI performed 30 h from estrus with fresh semen. Results of Experiment I showed that treatment (D) improved (P < 0.05) ovulatory response in comparison to Groups (C) and (A). The fertilization rate was lower (P < 0.01) in Group D) than Group (A). Also the proportion of transferable embryos was lower in Group (D) in comparison to all the other treatments (P < 0.01). Group A gave the best production of embryos (7.3/ewe; 89.0% transferable). In Experiment II, combined mating plus AI improved fertilization rate (80.3%) compared to both mating (P < 0.01) and AI (P < 0.02) alone.     

Effect of alfaprostol on postpartum reproductive efficiency in brahman cows and heifers

Brahman cows (n = 54) and heifers (n = 18) were randomly allotted by calving date, sex of calf and age to one of four treatment groups. Group 1 received no treatment (control), Group 2 received 5 mg alfaprostol (AP) i.m. on Day 21 postpartum, Group 3 received 5 mg AP i.m. on Day 32 postpartum and Group 4 received 5 mg AP i.m. on both Days 21 and 32 postpartum. Blood samples were collected via tail vessel puncture at 30 min-intervals for 8 h from half the animals in each group on Days 21 and 32 postpartum, with AP injection administered 2 h after sampling had begun. All cows were bled at weekly intervals. Samples were processed to yield serum and stored at -20 degrees C until assayed for luteinizing hormone (LH) or progesterone (P(4)). All cattle were maintained with epididymectomized marker bulls and were artificially inseminated (A.I.) at first estrus. Serum P(4) was below 1 ng/ml prior to AP treatment in all animals and did not differ (P > 0.10) between treatments. Alfaprostol treatment affected mean postpartum interval (from parturition to return to standing estrus and subsequent corpus luteum formation with serum progesterone concentrations > 1 ng/ml; P < 0.08). The control group (84.8 +/- 7.9 d) did not differ from Group 2 (86.3 +/- 11.1 d) or Group 3 (66.7 +/- 5.5 d) but did differ (P < 0.09) from Group 4 (65.1 +/- 6.4 d). Cattle injected on Day 32 had a shorter (P < 0.01) postpartum interval than those not receiving treatment on that day (65.9 +/- 4.2 vs 85.7 +/- 6.8 d). Pregnancy rate was affected (P < 0.05) by AP treatment. The control group (72.2%) did not differ (P > 0.10) from any group but, Group 2 (50.0%) was lower (P < 0.04) than Group 3 (83.3%) and (P < 0.02) Group 4 (88.9%). Cattle treated on Day 32 (Groups 3 and 4) had a higher (P < 0.02) pregnancy rate (86.1%) than those not treated on Day 32 (Groups 1 and 2; 61.1%). Serum LH was affected by day (P < 0.0003) and treatment by day (P < 0.07) but not by time (P > 0.10). Treatment Group 3 (P < 0.08) and Group 4 (P < 0.0003) mean LH concentrations differed between Days 21 and 32 postpartum. Cattle receiving AP treatment on Day 32 postpartum had a higher (P < 0.04) cumulative frequency of return to estrus by 100 days postpartum than nontreated cattle.     

Endocrine events during the periestrous period and the subsequent estrous cycle in ewes after estrus synchronization

The aim of the present study was to investigate the endocrinology of the periestrus period and that of the subsequent estrous cycle in ewes synchronized during the breeding season. Animals were treated for 14 days with either MAP intravaginal sponges or subcutaneous progesterone implants, followed by administration of 500 IU PMSG at the time of withdrawal. The time to estrus occurrence following progestagen withdrawal differed significantly between groups (45.3+/-2.7h for the MAP and 21.5+/-1.2h for the implant group, P<0.001). Estradiol levels around estrus did not differ between groups, but a significant difference was detected for the interval from peak estradiol to estrus, with a shorter interval for the implant group (26.7+/-0.7 and 2.7+/-0.9h, P<0.001). Progesterone implants shortened the interval from removal to LH surge, compared to the MAP group (31.2+/-4.4 and 56.5+/-3.6h, respectively, P<0.05). An earlier response was also observed for the interval from estradiol peak to LH peak in the implant group (12.1+/-3.3 and 37+/-2h, respectively, P<0.005), but no difference was observed for the interval from estrus to LH surge. Progesterone levels, particularly during the Days 6 to 10 of the subsequent estrous cycle were significantly higher (P<0.05) in the implant group. It is concluded that the kind of progesterone treatment may affect the time of estrus and the LH peak as well as the progesterone levels of the subsequent cycle.     

Serum hormone profiles and return to estrus in early-postpartum spring-lambing ewes treated with somatostatin

After parturition, 10 mature spring-lambing fine-wool ewes producing twins were allotted to one of two treatments. Five ewes received sterile saline (i.v.) twice daily on Days 12 to 15 post partum (PP) while 5 ewes were treated similarly except each injection contained 500 mug somatostatin (SRIF). Jugular blood samples were collected at 15-min intervals for 1 h before to 3 h after morning treatment on Days 12 and 15 PP. Animals were observed twice daily for signs of estrus using vasectomized rams beginning on Day 31 PP and continuing until ewes returned to estrus. Interval from parturition to estrus (mean +/- SEM) was similar (P > 0.40) in ewes receiving SRIF (119 +/- 6.2 d) and in control ewes (113 +/- 6.2 d). Ewes receiving 500 mug SRIF had lower (P < 0.10) serum insulin during the first 45 min after treatment on Day 12 PP; however, on Day 15 PP, serum insulin did not differ (P > 0.40) between treatment groups. Serum growth hormone (GH) did not differ (P > 0.40) between treatment groups 1 h before treatment on Day 12 PP; however, ewes treated with SRIF had lower (P < 0.05) GH levels before treatment on Day 15 PP than control ewes (4.4 and 9.9 +/- 1.5 ng/ml, respectively). After administration of SRIF, serum GH was higher (P < 0.05) in SRIF-treated ewes than in controls (8.2 and 5.3 +/- 2.7 ng/ml, respectively) on Day 12 PP but no differences (P > 0.80) were noted between treatment groups on Day 15 PP. These data indicate that 500 mug SRIF given twice daily from Days 12 to 15 PP neither lowered serum GH nor influenced return to estrus in lactating fine-wool ewes.     

Can audio–visual or visual stimuli from a prospective mate stimulate a reproductive neuroendocrine response in sheep?

Stimuli from a prospective mate increase the secretion of luteinising hormone (LH) in sheep. This 'male effect' in ewes and 'female effect' effect in rams is predominantly mediated by olfactory signals, though it is thought that non-olfactory signals play synergistic or substitutive roles. In this study, we tested whether exposure to visual or audio-visual stimuli from a prospective mate would stimulate an increase in LH secretion in ewes (Experiment 1) and rams (Experiment 2). In Experiment 1, groups of eight Merino ewes were exposed to one of three stimuli midway through a frequent blood-sampling regimen: full ram contact, still images of rams, a video of ewes and rams mating. Control ewes (n = 8) were completely isolated from rams. Exposure to still images of rams appeared to stimulate an increase in mean LH concentrations (P < 0.05) and tended to increase LH pulse frequency (P < 0.1), but the response was significantly smaller than that observed in ewes exposed to rams (P < 0.01). Audio-visual stimuli had no effect on any parameters of LH secretion (P > 0.1). In Experiment 2, Merino rams were allocated to either an Exposure (n = 7) or a Control (n = 7) group. Exposure rams underwent two exposure periods midway through a frequent blood-sampling regimen; exposure to still images of ewes and audio recorded during mating of ewes and rams (audio-visual exposure); exposure to oestrous ewes (ewe exposure). Control rams were sampled at the same frequency but remained isolated from ewe stimuli. Exposure of rams to the audio-visual stimuli did not affect any parameters of LH secretion (P > 0.1). In contrast, exposure to oestrous ewes increased LH pulse frequency (P < 0.05) and advanced the onset of the next LH pulse (P < 0.05). In conclusion, visual signals appear to be involved in eliciting the neuroendocrine response of ewes to rams and are of greater importance to this phenomenon in ewes (male effect) than rams (female effect). However, overall the visual and audio-visual signals used in this study were far less effective than stimulus animals, suggesting that these stimuli are less important than olfactory signals, or a combination of olfactory and audio-visual signals.     

Effect of transport on pulsatile and surge secretion of LH in ewes in the breeding season.

The aim of this study was to elucidate the mechanism(s) involved in stress-induced subfertility by examining the effect of 4 h transport on surge and pulsatile LH secretion in intact ewes and ovariectomized ewes treated with steroids to induce an artificial follicular phase (model ewes). Transport caused a greater delay in the onset of the LH surge in nine intact ewes than it did in ten ovariectomized ewes (intact: 41.0 +/- 0.9 h versus 48.3 +/- 0.8 h, P < 0.02; ovariectomized model: 40.8 +/- 0.6 h versus 42.6 +/- 0.5 h, P < 0.02). Disruption of the hypothalamus-pituitary endocrine balance in intact ewes may have reduced gonadotrophin stimulation of follicular oestradiol production which had an additional effect on the LH surge mechanism. In the ovariectomized model ewes, this effect was masked by the exogenous supply of oestradiol. However, in these model ewes, there was a greater suppression of maximum LH surge concentrations (intact controls: 29 +/- 4 ng ml-1 versus intact transported 22 +/- 5 ng ml-1, P < 0.02; ovariectomized model controls: 35 +/- 7 ng ml-1 versus model transported 15 +/- 2 ng ml-1, P < 0.02). Subsequent exposure to progesterone for 12 days resulted in the resumption of a normal LH profile in the next follicular phase, indicating that acute stress leads to a temporary endocrine lesion. In four intact ewes transported in the mid-follicular phase, there was a suppression of LH pulse amplitude (0.9 +/- 0.3 versus 0.3 +/- 0.02 ng ml-1, P < 0.05) but a statistically significant effect on pulse frequency was not observed (2.0 +/- 0.4 versus 1.7 +/- 0.6 pulses per 2 h). In conclusion, activation of the hypothalamus-pituitary-adrenal axis by transport in the follicular phase of intact ewes interrupts surge secretion of LH, possibly by interference with LH pulsatility and, hence, follicular oestradiol production. This disruption of gonadotrophin secretion will have a major impact on fertility.     

Duration of the breeding season and response to reproductive manipulation in five breeds of sheep under northern prairie conditions

The breeding season was 157, 154, <126, 210 and 217 days for Rambouillet, Columbia, Suffolk, Rambouillet x Finnish Landrace and Columbia x Finnish Landrace ewes respectively. Treatment of cyclic ewes with pregnant mare serum gonadotropin (PMSG) (500 IU), following a 12-day treatment with progestin-containing intravaginal sponges, did not affect fertility, but did decrease the time from sponge removal to estrus, (control 48.0 +/- 3.1 hr; PMSG 39.4 +/- 1.8 hr) to the preovulatory surge of LH (control 52.7 +/- 2.8 hr; PMSG 39.0 +/- 1.7 hr) and FSH (control 52.3 +/- 2.9 hr; PMSG 42.8 +/- 1.6 hr) and caused an elevation of serum LH levels prior to the preovulatory surge (control 1.25 +/- 0.18 ng/ml; PMSG 2.31 +/- 0.22 ng/ml). Exposure of the purebred ewes to 18 hours of daylight in January, decreasing by 30 minutes a week subsequently, counteracted the seasonal reduction in the number of ewes lambing following induced breeding under natural daylight in May. Prolificacy was greatest in crossbred ewes and their fertility was not affected by season. Gestation period was longer for fall-bred ewes and varied with breed.     

Effect of recombinant alpha interferons on fertility and interestrous interval in sheep

In Experiment 1, 12 unmated cyclic ewes received twice-daily intrauterine injections on Days 12 to 14 of one of the following treatments: 1) ovine conceptus secretory proteins (oCSP) containing 25 mug of ovine trophoblast protein-1 (oTP-1) as determined by RIA; 2) 25 or 50 mug recombinant human interferon alpha1 (rhlFN); or 3) 1500 ug of serum proteins (oSP) from a Day-16 pregnant ewe (estrus = Day 0) per uterine horn. Ewes receiving oCSP had longer interestrous intervals (27 +/- 2 days; P<0.05) than ewes receiving oSP (17 +/- 2 days). Ewes receiving either dose of rhlFN had an interestrous interval of 16 +/- 2 days which did not differ (P>0.10) from that of oSP-treated ewes. In Experiment 2, 59 normally cycling ewes, mated on Day 0, received twice-daily intramuscular injections of either 2 mg recombinant bovine interferon alpha1 (rblFN) or placebo on Days 12 to 15 post estrus. On Day 16, pregnancy was confirmed by flushing a morphologically normal conceptus from the uterus. Pregnancy rates for rblFN-treated (80%) and placebo-treated (62%) ewes were not different (P>0.10). Uterine flushings and conceptus-conditioned medium were assayed for oTP-1. Total oTP-1 in conceptus-conditioned culture medium was higher (P<0.02) when conceptuses were from placebo-treated (104 +/- 14 mug/conceptus) than from rblFN-treated (56 +/- 12 mug/conceptus) ewes; while total oTP-1 in uterine flushings was similar (P>0.10) for placebo-treated (132 +/- 15 mug/conceptus) and rblFN-treated (147 +/- 17 mug/conceptus) ewes. The interval from mating to subsequent estrus following conceptus removal was 31 +/- 1 and 28 +/- 1 days for pregnant ewes treated with rblFN and placebo, respectively. Interestrous intervals for nonpregnant ewes were longer (P<0.02) for rblFN-treated (27 +/- 3 days) than for placebo-treated (18 +/- 2 days) ewes.     

Follicular growth, endocrine response and embryo yields in sheep superovulated with FSH after pretreatment with a single short-acting dose of GnRH antagonist

The objective of this study was to characterize follicular development, onset of oestrus and preovulatory LH surge, and in vivo embryo yields of sheep superovulated after treatment with a single dose of 1.5mg of GnRH antagonist (GnRHa). At first FSH dose, ewes treated with GnRH antagonist (n=12) showed a higher number of gonadotrophin-responsive follicles, 2-3mm, than control ewes (n=9, 13.5+/-3.8 versus 5.3+/-0.3, P<0.05). Administration of FSH increased the number of >or=4mm follicles at sponge removal in both groups (19.3+/-3.8, P<0.0005 for treated ewes and 12.7+/-5.4, P<0.01 for controls). Thereafter, a 25% of the GnRHa-treated sheep did not show oestrous behaviour whilst none control sheep failed (P=0.06). The preovulatory LH surge was detected in an 88.9% of control ewes and 66.7% of GnRHa-treated sheep. A 77.8% of control females showed ovulation with a mean of 9.6+/-0.9 CL and 3.3+/-0.7 viable embryos, while ewes treated with GnRHa and showing an LH surge exhibited a bimodal distribution of response; 50% showed no ovulatory response and 50% superovulated with a mean of 12.2+/-1.1 CL and 7.3+/-1.1 viable embryos. In conclusion, a single dose of GnRHa enhances the number of gonadotrophin-dependent follicles able to grow to preovulatory sizes in response to an FSH supply. However, LH secretion may be altered in some females, which can affect the preovulatory LH surge and/or can weak the terminal maturation of ovulatory follicles.     

Influence of the type of energy supply on lh secretion, follicular growth and response to estrus synchronization treatment in feed-restricted suckler beef cows

The present experiment aimed to compare the efficiency of supplementation (+17.5 MJ Net Energy/d starting 47 +/- 4 days after calving) with concentrate (CS, maize grain, n = 10) or with forage (FS, maize silage, n = 10) in estrus-synchronized (Norgestomet implant 10 days inserted 60 +/- 4 days postpartum + PMSG at implant removal) beef cows previously restricted (47 MJ Net Energy/d, 785 g CP/d, 70% of requirements). The type of diet had no significant effect on basal LH concentrations (CS: 0.18 +/- 0.12 vs FS: 0.11+/- 0.02 ng/mL), LH pulse frequency (CS : 0.7 +/- 0.3 vs FS: 0.8 +/- 0.2 pulse/10 h), LH pulse amplitude (CS: 0.55 +/- 0.50 vs FS : 0.62 +/- 0.50 ng/mL) or estradiol (E2) concentrations (CS: 3.3 +/- 0.8 vs FS: 4.6+ /- 0.8 pg/mL) 13 days after the beginning of energy supplementation. No differences between CS and FS cows were observed for the number of small, medium and large follicles nor on the size of the largest follicle from 11 days before implant insertion to implant removal (IR). After IR, an LH surge was observed in 2 of the CS and 4 of the FS cows. The type of energy supplementation had no significant effect on LH (CS: 0.16 +/- 0.06 ng/mL vs FS 0.48 +/- 0.06 ng/mL; P > 0.05) or on estradiol concentrations (CS : 7.8 +/- 0.2 vs FS : 8.9 +/- 0.2 pg/mL, P > 0.10) measured hourly from 29 to 49 h after IR. Cows that ovulated after IR tended to have higher E2 concentrations than cows that did not ovulate (9.4 +/- 0.2 vs 6.3 +/- 0.2 pg/mL, P = 0.08). Similar ovulation and pregnancy rates were observed in CS and FS cows (CS: 6/10 vs FS: 7/10 and CS: 6/10 vs FS: 5/10 respectively, P > 0.05). To conclude, energy supplementation with forage was as effective as energy supplementation with concentrate to influence follicular growth, ovulation and pregnancy percentage after estrus synchronization treatment in diet-restricted beef cows.     

Effects of oestradiol on LH, FSH and prolactin in ovariectomized ewes

This study was conducted to test the hypothesis that the rate (dose/time) at which oestradiol-17 beta (oestradiol) is presented to the hypothalamo-pituitary axis influences secretion of LH, FSH and prolactin. A computer-controlled infusion system was used to produce linearly increasing serum concentrations of oestradiol in ovariectomized ewes over a period of 60 h. Serum samples were collected from ewes every 2 h from 8 h before to 92 h after start of infusion, and assayed for oestradiol, LH, FSH and prolactin. Rates of oestradiol increase were categorized into high (0.61-1.78 pg/h), medium (0.13-0.60 pg/h) and low (0.01-0.12 pg/h). Ewes receiving high rates of oestradiol (N = 11) responded with a surge of LH 12.7 +/- 2.0 h after oestradiol began to increase, whereas ewes receiving medium (N = 15) and low (N = 11) rates of oestradiol responded with a surge of LH at 19.4 +/- 1.7 and 30.9 +/- 2.0 h, respectively. None of the surges of LH was accompanied by a surge of FSH. Serum concentrations of FSH decreased and prolactin increased in ewes receiving high and medium rates of oestradiol, when compared to saline-infused ewes (N = 8; P less than 0.05). We conclude that rate of increase in serum concentrations of oestradiol controls the time of the surge of LH and secretion of prolactin and FSH in ovariectomized ewes. We also suggest that the mechanism by which oestradiol induces a surge of LH may be different from the mechanism by which oestradiol induces a surge of FSH.     

Ewe effect: endocrine and testicular changes in experienced adult and inexperienced young Corriedale rams used for the ram effect

Changes in LH, FSH, and testosterone concentrations, testicular firmness and resilience, and scrotal circumference were monitored in 16 Corriedale rams (8 experienced adult and 8 inexperienced young) for 20 days during which they were used to stimulate ewes. The experiment was conducted during November (mid-non-breeding season). Increases (P<0.05) were observed in LH and testosterone concentrations and in testicular firmness and resilience during the first 4 days when rams were in permanent contact with estrual ewes. During the following 13 days, when rams were in contact with non-estrual ewes (i.e. initially estrual ewes were no longer in estrus), LH and testosterone concentrations decreased. When initially anestrous ewes exhibited estrus 17 to 20 days later, concentrations of testosterone increased. Testicular firmness and resilience remained high throughout the period. We conclude that: (1) rams used to stimulate anestrous ewes show an increase in LH and testosterone concentrations beginning at 12 h after joining, and greater concentrations are maintained while estrual ewes and mating are allowed; and (2) estrous and mating activity are probably the most important stimuli for the increase in hormone concentrations.