Effect of GnRH administration, combined with the ram effect, on the occurrence of ovulation and pregnancy during the transition period from anoestrus in crossbred ewes

The effect of a GnRH analogue (buserelin) combined with the ram effect on the reproductive efficiency of ewes was investigated in 105 cross-bred fat tailed ewes, during the transition period from anoestrus to the natural breeding season. Plasma progesterone concentration was used in the assessment with regard to ovulation and pregnancy. Ewes were maintained on natural pastures composed of medium to low quality forages, and received supplementation (40% alfalfa hay: 60% wheat straw) ad libitum, plus 100–300 g barley grain per head per day. Ewes were isolated from the rams for at least two months and then kept in close proximity of the rams for one week, before the introduction of the rams. The ewes were randomly divided into three equal groups (n = 35 per group). With the introduction of the rams into the flock (day 1) one group was considered as the control and the other two groups were treated with 4.2 μg (low dose) and 8.4 μg (high dose) buserelin on days 5 and 19, respectively. Blood samples were collected on days 5, 12, 26 and 120 after ram introduction for determining the plasma progesterone levels. On day 12 (one week after treatment) the high dose GnRH group recorded significantly lower plasma P4 concentrations (P < 0.05), compared with the control group (1.0 ± 0.1 ng/ml versus 2.4 ± 0.4 ng/ml). On the same day the low GnRH dose group recorded intermediate P4 concentrations, recording no significant differences with the other two groups. The high dose group recorded a significantly (P < 0.05) higher proportion of non-ovulated ewes (61.8%), compared to the control (32.3%) and low dose (31.4%) groups on day 12 of the study. At days 5 and 26 these differences were not significant, but the proportion of non-ovulated ewes was higher in the high dose buserelin treatment group. The percentage of pregnant (plasma P4 > 2.5 ng/ml) and non-pregnant (plasma P4 ≤ 2.5 ng/ml) ewes at day 120 of the study was not statistically different between the treatment groups. The pregnancy rate was highest in the control group (97.1%), when compared to the treated ewes (94.3% and 88.6% in low dose and high dose treatment groups, respectively). Treatment with buserelin combined with the male effect during the breeding season negatively affected the plasma P₄ concentration, reducing the reproductive performance of the ewe treatment groups.     

Response of anestrous ewes to the ram effect after follicular wave synchronization with a single dose of estradiol-17beta

Anestrous ewes respond to the introduction of rams with either an ovulation within 2-3 days that may be followed by luteal phases of normal or short length, with delayed ovulations (5-6 days later), or with the luteinization of follicles. The aim of this work was to study the relationship between the growth status of the largest follicle present when rams are introduced and the type of ovarian response in non-treated ewes and in ewes treated with estradiol-17beta before ram introduction. Thirteen anestrous Corriedale ewes were divided into 2 groups: E2 (n = 7) and C (n = 6). The E2 ewes received a single dose of 50 microg estradiol-17beta 5 days before the introduction of the rams to synchronize the onset of their follicle waves, while C ewes remained untreated. When the rams were introduced, all E2 ewes had the largest follicle in a growing stage in contrast with the C ewes (3 out of 6; P < 0.05). Five C and 4 E2 ewes ovulated after the introduction of the rams (Day 3.4 +/- 0.4 for C vs. 4.8 +/- 0.3 for E2 ewes, respectively, P < 0.05). Only one ewe from each group developed a normal luteal phase: 4 C and 3 E2 ewes had short luteal phases. One C ewe and 2 E2 ewes had short luteal phases originating from follicles that did not ovulate. After the first luteal phase, all ewes returned to anesirus without a second ovulation or luteal phase. The remaining E2 ewe did not ovulate or show any changes in progesterone serum concentrations. We conclude that the growth status of the largest follicle alone does not determine the ovarian responding pattern of anestrous ewes to the ram effect.     

Reproductive response and LH secretion in ewes treated with melatonin implants and induced to ovulate with the ram effect

Two experiments were conducted to examine the effects of treating seasonally anoestrous ewes with melatonin before ram introduction on reproductive response, and on LH secretion in anoestrous ewes induced to ovulate by rams.In Experiment 1, a total of 667 ewes from three flocks involving Merino (Flock 1, N = 149), Merino entrefino (Flock 2, N = 325) and Rasa Aragonesa (Flock 3, N = 203) breeds were used. Within each flock, ewes isolated from rams since the previous lambing were assigned at random to receive melatonin implants of Regulin (75, 175 and 105 in Merino, Merino entrefino and Rasa Aragonesa flocks, respectively) or to serve as untreated controls (74 in Merino, 150 in Merino entrefino and 98 in Rasa Aragonesa flocks). Fertile rams were introduced into all flocks 5 weeks after implantation in March (Flocks 1 and 2) or April (Flock 3), and remained with the ewes for a 50 day mating period. Percentage of ewes with luteal activity at ram introduction did not differ between melatonin treated and control ewes in any flock. There were no significant differences in either the mean interval from ram introduction to lambing or the distribution of lambing. Implantation with melatonin resulted in an improvement of prolificacy in all three flocks, although this only reached statistical significance in the Merino flock (1.15 vs. 1.03 in treated and control ewes, respectively, P < 0.05). Fertility was increased significantly (P < 0.05) in the Merino entrefino flock (64.5% in treated vs. 51.3% in control ewes).In Experiment 2, two trials were undertaken utilizing a total of 63 ewes. Trial 1 involved 24 mature Manchega ewes and Trial 2 involved 39 Merino ewe lambs. Half of the animals in each trial received a Regulin implant on 28 February (Trial 1) or 12 March (Trial 2) and the remaining half acted as controls. Rams were introduced 5 weeks after implantation and remained with the ewes for a 25 day period. In both trials, anoestrous ewes at ram introduction were bled at 20 min intervals for 3 h before and 5 h after ram introduction and then at 3 h intervals over the next 24 h for assessment of plasma concentrations of LH. Secretion of LH before or following introduction of rams was not affected by melatonin. Both treated and control anoestrous ewes in each trial responded to introduction of rams with an increase in the frequency of the LH pulses (P < 0.05), but no significant changes were detected in pulse amplitude or mean levels of LH. A preovulatory surge of LH was detected between 8 and 26 h after ram introduction, but neither mean interval from ram introduction to the peak of LH surge, nor the magnitude of the LH peak, was influenced by melatonin treatment.Results from this study show that: (1) melatonin implants administered during early seasonal anoestrus have the potential to improve reproductive performance in Spanish breeds of sheep, but the response is conditioned by breed, management system and environmental factors; (2) melatonin did not modify the secretion of LH in anoestrous ewes induced to ovulate by the ram effect under our experimental conditions.     

Effects of the presence of rams during pregnancy on lambing performance in ewes

Ewes of the Préalpes-du-Sud breed (n=112) were mated with fertile rams and were used to investigate the effect of the presence of vasectomized rams during pregnancy on reproductive outcomes. Ewes in the control group (n=56) were isolated from rams during the whole period of pregnancy, whereas those in the experimental group (n=56) were kept with vasectomized rams from day 10 post-mating until lambing. At day 10 post-mating, a series of blood samples was collected every 15 min for 8 h from five control ewes and five experimental ewes to determine their patterns of LH secretion. The introduction of the ram was associated with a rapid increase of pulsatile LH release. The lag between the introduction of the ram and the onset of the first episodic LH release was less than 15 min. The mean(±sem) number of LH pulses/4 h after the introduction of the ram (2.8±0.4) was significantly higher (P<0.01) than that observed/4 h before the introduction of the ram (1.4±0.2). Although more ewes were pregnant in the control group (87.5%) than in the ram-exposed group (82.1%), the difference was not significant. The presence of rams did not affect gestation length (145.8 days), overall lamb mortality (3.5%) or birth weights of single (3.96 kg), twin (3.24 kg) or triplet (2.59 kg) lambs. The proportion of ewes with multiple births in the control group (69.4%) was significantly greater (P<0.05) than that in the ram-exposed ewes (47.8%). The ewes in the control group had significantly more (P<0.01) twin lambs born alive (72.3%) than the ewes in the ram-exposed group (50.0%). In conclusion, the presence of vasectomized rams during early pregnancy affected the incidence of multiple births but did not affect pregnancy rate or gestation length. The altered fertility of ewes exposed to vasectomized rams may reflect changes in embryonic loss during early pregnancy.     

Progesterone exposure of the preovulatory follicle in the seasonally anestrous ewe alters the expression of angiogenic growth factors in the early corpus luteum

Gonadotrophin releasing hormone (GnRH)-induced ovulation in seasonally anestrous ewes is associated with a high incidence of defective corpora lutea (CL), which can be completely eliminated by priming ewes with progesterone before GnRH treatment, but the physiological basis of this has remained elusive. This study tested the hypothesis that progesterone priming eliminates defective luteal function by altering the expression of Vascular Endothelial Growth Factor (VEGF), its receptor VEGFR-2, and angiopoietin (ANG)-1, ANG-2 and their receptorTIE-2 in the early CL. Fifteen seasonally anestrous ewes were treated by i.m. injection with 20 mg of progesterone 3 days before the start of GnRH treatment, while another 15 animals served as controls. Intravenous injections of 500 ng GnRH were given to all the ewes every 2 h for 28 h, followed by a 300 μg GnRH bolus injection to synchronize the preovulatory luteinizing hormone (LH) surge. Corpora lutea were collected 1, 2 and 4 days after ovulation and analyzed for protein and mRNA expression of VEGF, VEGFR-2, ANG-1, ANG-2 and Tie-2 using Western Immunoblotting and in situ hybridization. VEGF, VEGFR-2 and ANG-1 expression was significantly higher (P ≤ 0.05) in the CL of progesterone-primed animals compared to non-primed ones. However, no differences were observed in the ANG-2 or Tie-2 expression levels between the two treatment groups. These data suggest that progesterone priming of the preovulatory follicle alters the expression of some angiogenic growth factors in the early CL, leading to greater vascular stability and thereby normal luteal function.     

Corpus luteum function in ewes stimulated by rams

In late February Dorset rams were introduced (day = 0) to 40 mature Romney ewes that were observed by laparoscopy to be anovular. The ovaries of 20 of these ewes were examined by laparoscopy every second day while the remaining 20 ewes served as unoperated controls. Jugular blood samples were taken daily and plasma progesterone concentrations assayed to provide information on the functional status of any corpora lutea (CL) arising from ovulations stimulated by introduction of the rams. Eighty-five percent (-17/20) of the ewes that were repeatedly laparoscoped had ovulated within 4 days of ram introduction and premature regression of the CL had occurred between days 4 and 8 in 8 ewes and days 6 to 10 in 2 ewes. A second ovulation was observed after or during the premature regression of the first CL and this subsequent CL was maintained for the normal duration. The prematurely regressing CL produced a small peak in progesterone concentration on days 4 to 5 but the concentrations declined on days 6 to 7. In the unoperated ewes 50% (-10/20) appeared, from the progesterone profiles, to have ovulated by day 4 and half of these appeared to have premature CL regression. The interval from introduction of the ram to first oestrus was 23 days in ewes with premature regression of the CL and 19 days in ewes ovulating within 4 days but having no premature regression. From the results it was concluded that the premature regression of the CL is the cause of the delayed interval from ram introduction to first oestrus in Romney ewes and is a major factor contributing to the two peaks of oestrous activity observed after ram introduction.     

Effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus

The effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus were investigated in two experiments. In the first experiment, the introduction of rams induced an increase in the levels of LH in entire ewes. The mean levels increased from 0.68 +/- 0.04 ng/ml (mean +/- s.e.m.) to 4.49 +/- 1.32 ng/ml within 20 min in ewes not treated with progesterone (n = 10). In ewes bearing progesterone implants that provided a peripheral concentration of about 1.5 ng progesterone per millilitre plasma, the LH response to the introduction of rams was not prevented, but was reduced in size so that the concentration was 1.38 +/- 0.15 ng/ml after 20 min (n = 5). Progesterone treatment begun either 2 days before or 6 h after the introduction of rams and maintained for 4 days prevented ovulation. In the second experiment ovariectomized ewes were used to investigate further the mechanism by which the ram evoked increases in tonic LH secretion. In ovariectomized ewes treated with oestradiol implants, the introduction of rams increased the frequency of the LH pulses and the basal level of LH. In the absence of oestradiol there was no significant change in pulse frequency but a small increase in basal levels. Progesterone again did not prevent but reduced the responses in ewes treated with oestradiol. It is suggested that following the withdrawal of progesterone treatment, the secretion of LH pulses in response to the ram effect would be dampened. This effect could be a component of the reported long delay between the introduction of rams and the preovulatory surge of LH in ewes treated with progesterone. Continued progesterone treatment prevented ovulation, probably by blocking positive feedback by oestradiol.     

Effects of Melatonin,Progestagens, and the Ram on Out-of-Season Reproduction in Swedish Landrace Finewool Sheep

One hundred twenty-one Swedish Landrace Finewool ewes were treated with progestagen sponges (P), teaser ram stimulation (R), or melatonin implants plus teaser ram stimulation (M) in preparation for breeding with whole rams in August. Blood progesterone analyses from ewes in the R and M groups gave no evidence of luteal activity before the introduction of teaser rams. There were no significant differences between treatments for pregnancy rate (~90%). The P group had the most compact lambing season, while median breeding dates for M and R groups were delayed by one cycle. In those groups, the introduction of breeding rams was later found to have been too late. M and R differed significantly for probable conception date but not for lambing dates. Circa 30% of M ewes did not have a short 6 day ovulation cycle after the first ovulation, which resulted in a less concentrated lambing season than the other methods. Although no significant differences in litter size were seen among the 3 treatments, M had the highest group average, 2.25. The ewes in this study were not in very deep anestrous in the middle of August. This supports the conclusion that treatment with exogenous hormones is not necessary to breed Swedish Landrace Finewool ewes successfully in late August/early September.     

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.     

Effects of lactation status, progestogen and ram exposure on response to cloprostenol in ewes during the anestrous season

Progestogen pretreatment and introduction of rams were used to prepare 432 Rasa aragonesa ewes for synchronization of estrus with prostaglandin (PG) during anestrus. The experiment was a 3 x 3 x 2 factorial with lactation status, ram/progestogen treatment and treatment with pregnant mare's serum gonadotropin (PMSG; 250 IU) at injection of PG as main effects. At ram introduction (Day 0), ewes were dry (Group 1), weaned (Group 2) or suckling a lamb (Group 3). They received either norgestomet implants for 12 d (Days -12 to Day 0 (Group A), ram introduction for 12 d (Days 0 to 12) (Group B) or both (Group C). Half the ewes received PMSG with PG (100 mug Cloprostenol) on Day 12. Pregnancy rate was higher at first service in dry (Group 1; 42%) than in recently-weaned ewes (Group 3; 24% and Group 2; 31%; P<0.01). Occurrence of estrus and conception and pregnancy rates to first service were higher (P<0.01) in ewes previously exposed to rams (Groups B and C) than in ewes treated only with progestogen (Group A). There were no effects of PMSG, no interactions among the three variables and no differences in prolificacy (1.12 at first service).     

Synchronization of estrus with short- and long-term progestagen treatments and the use of GnRH prior to short-term progestagen treatment in ewes

The objective of the present study was to determine the efficacy of the synchronization of estrus using short- and long-term progestagen treatments in ewes at the onset of the breeding season, and to evaluate the effect of the exogenous GnRH administration immediately prior to short-term progestagen treatment on the reproductive performance. A total of 240 Tahirova cross-bred ewes, aged 18–24 months, and 40 rams, aged 2–4 years-old, were used in the trial. Ewes were divided equally into 3 groups (n = 80 per group). Intravaginal progestagen sponges containing FGA (30 mg) were inserted in the ewes for 7 d in the FGA1 (short-term) and GnRH treatment groups, and for 12 d in the FGA2 group (long-term). The ewes in the GnRH group received 10.5 μg busereline acetate i.m. at the time of sponge insertion. Tiaprost tromethamol (PGF_2α; 0.294 mg) and eCG (400 IU) were injected i.m. on the 6th day of progestagen treatment in the GnRH and FGA1 groups, and on the 11th day in the FGA2 group following sponge insertion. All ewes were hand-mated once at the detection of estrus. The estrous response, fertility rate, multiple birth rate and litter size recorded was 88.7, 87.3, 51.6% and 1.6 in the FGA1 group, 92.5, 71.6, 50.9% and 1.5 in the FGA2 group, and 96.2, 89.6, 71.0% and 1.8 in the GnRH group, respectively. No significant difference in estrous response between the groups was recorded, but the fertility rate in the FGA1 and GnRH groups was significantly (P < 0.05) higher than in the FGA2 group. The occurrence of multiple births and litter sizes were significantly (P < 0.05) higher in the GnRH group, compared to both the FGA1 and FGA2 groups, with the number of single lambs being significantly (P < 0.05) higher in the FGA1 (48.4%) and FGA2 (49.0%) groups than in the GnRH (29.0%) group. However, the differences recorded between any of the groups in terms of the number of twin and triplet lambs were insignificant. In conclusion, it can be said that estrous synchronization using the 12-d-FGA-eCG-PGF_2α regimen could be replaced with the 7-d-FGA-eCG-PGF_2α regime in sheep at the onset of the breeding season. However, the combination of GnRH with the latter regimen (7-d-GnRH-FGA-eCG-PGF_2α) increased the multiple birth rate and litter size in the ewes.     

Effect of early and late exposure to estrual ewes on ram sexual performance classifications

This study was conducted to determine whether exposure of ram lambs to estrual ewes during their first autumn and again as adults just before serving capacity tests (SCT) affected the outcome of the sexual performance tests. Treatments were either early exposure of Polypay ram lambs (i.e., 7-8-mo-old rams with ewes for 17 d [n=30] or no early exposure [n=30]), and late exposure (i.e., 16-19-mo-old rams with estrual ewes for 3 d) or no exposure to estrual ewes in a 2x2 factorial arrangement. Three serving capacity tests were conducted immediately after the early exposure period for individual ram lambs that were exposed to ewes early. Three sham sexual performance tests (i.e., four ram lambs placed in test pens for 30-min without ewes) were conducted with ram lambs that were not exposed to ewes early. All rams were evaluated during nine 30-min serving capacity tests over a 2-mo period at 16-19 mo of age to determine sexual performance. Prior to serving capacity tests, one half of the rams from each early exposure treatment were exposed to estrual-induced ewes for 3 d. Specific sexual behaviors (e.g., sniffs, flehmens, foreleg kicks, vocalizations, mount attempts, mounts, and ejaculations) were recorded during serving capacity tests. Number of sniffs, flehmens, foreleg kicks, vocalizations, and mount attempts were summed without estimating the value of importance and analyzed as courtship behaviors. Sexual performance data were analyzed with Mixed model procedures for repeated measures. During serving capacity tests, the early exposed rams exhibited more courtships (40.3+/-8.0 versus 23.4+/-4.6; P<0.05; LSM+/-estimated SE), mounts (11.3+/-1.0 versus 7.7+/-0.9; P<0.01), and ejaculations (3.3+/-0.2 or 2.4+/-0.2; P<0.01) than rams not exposed to ewes as ram lambs, respectively. We conclude that early exposure of 7-8-mo-old ram lambs to estrual ewes improves sexual performance in serving capacity tests at 16-19 mo of age in most rams whereas, late exposure to estrual ewes for 3 d prior to serving capacity tests did not improve sexual performance scores.     

Endocrine and Ovarian Changes in Response to the Ram Effect in Medroxyprogesterone Acetate-primed Corriedale Ewes During the Breeding and Nonbreeding Season

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4–6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17β were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17β concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17β, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17β levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17β concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.     

Effects of recent sexual experience and melatonin treatment of rams on plasma testosterone concentration, sexual behaviour and ability to induce ovulation in seasonally anoestrous ewes

The aim of this study was to determine whether advancing the seasonal changes associated with rams by treatment with exogenous melatonin and allowing the rams previous sexual experience would increase the proportion of anoestrous ewes ovulating in early July. North Country Mule ewes (n = 225) were grouped by live body weight and body condition score and allocated randomly to the following treatments: (i) isolated from rams (control; n = 25); (ii) introduced to rams (treatment 2); (iii) introduced to rams that had mated with ewes during the previous 2 days (treatment 3); (iv) introduced to rams implanted with melatonin (treatment 4); and (v) introduced to rams that were implanted with melatonin and had mated with ewes during the previous 2 days (treatment 5). Treatments 2-5 were replicated (2 x 25 ewes) and two rams were introduced to each replicate group. Introductions began on 4 July and were completed by 11 July. The rams were withdrawn from the ewes after 8 days. Melatonin was administered as a subcutaneous implant (Regulin((R))) on 22 May and again on 20 June. Blood samples were taken from all rams to determine plasma melatonin and testosterone concentrations (19 samples in 6 h). The behaviour of the sheep was videotaped continuously during the first 3 h after the ram was introduced. Ovulation was detected by an increase in plasma progesterone concentrations from < 0.5 ng ml(-1) to > 0.5 ng ml(-1). Mean +/- SE plasma melatonin concentrations were 649.7 +/- 281.4 and 18.3 +/- 2.4 pg ml(-1) in rams with and without melatonin implants, respectively (P < 0.001). Melatonin implants also increased plasma testosterone concentrations from 4.30 +/- 1.88 to 10.10 +/- 1.10 ng ml(-1) (P < 0.01), the libido of the rams and the proportion of ewes that ovulated in response to the rams (43 and 56% (treatments 4 and 5) versus 24% (treatments 2 and 3)). In conclusion, implanting rams with melatonin before introducing them to seasonally anoestrous ewes increases the proportion of ewes that ovulate in response to introduction of a ram, but previous sexual experience of rams appears to have little or no effect.     

Short-term treatment with a controlled internal drug releasing (CIDR) device and FSH to induce fertile estrus and increase prolificacy in anestrous ewes

The objectives were to evaluate, in anestrous ewes, the effectiveness of a CIDR-G device (0.3 g progesterone) administered for 5 d to induce estrus; and FSH (Folltropin; 55 mg NIH-FSH-P1 equivalent) in saline:propylene glycol (1:4) 24 h before insert removal (Day 0), to increase ovulation rate and prolificacy. Ewes of mixed breeding were assigned at random to 3 treatments: control (C; n = 125), 5 d progesterone (P5; n = 257) and 5 d progesterone plus FSH (P5F; n = 271). Intact rams were joined at insert removal and ewes were observed every 24 h for 3 d. On Day 14, the ovulation rates of all ewes detected in estrus in the treated groups were determined using transrectal ultrasonography. Rams were removed on Day 26 to 31. Ewes were examined for pregnancy then, and again 20 to 25 d later to detect ewes that conceived to the second service period. Percentage of ewes marked by rams was higher in progesterone-treated (77%) than in C (20%; P < 0.01), but did not differ between P5 and P5F. The ovulation rate (1.95+/-0.04) did not differ due to FSH. Conception (68%) and pregnancy (52%) rates were higher in progesterone-treated (P < 0.01) than in C (0%) ewes. Estrous response varied quadratically with time after ram introduction, and the conception rate varied quadratically with the time of observation of onset of estrus. Over two service periods more progesterone-treated than C ewes lambed (65 vs 45%; P < 0.01). Lambs born per ewe exposed (0.7+/-0.1, 1.0+/-0.1, and 1.1+/-0.1 for C, P5 and P5F, respectively) was increased by progesterone (P < 0.05). Litter size to the first service period (1.59+/-0.04) and overall (1.54+/-0.03) did not differ among treatment groups. FSH-treated ewes tended to have more lambs (1.67+/-0.1) than did ewes receiving progesterone alone (1.5+/-0.1; P = 0.06) and than did ewes lambing to the second service period (1.5+/-0.1; P = 0.06). In summary, a 5-d progesterone pre-treatment of anestrous ewes induced estrous cycles and increased the pregnancy rates. A single injection of FSH only tended to increase litter size.     

Prior exposure of maiden ewes to rams enhances their behavioural interactions with rams but is not a pre-requisite to their endocrine response to the ram effect

In this study, we tested whether prior experience with rams would modify the behavioural and endocrine responses of maiden ewes to rams. During mid-anoestrus, sexually na?ve, maiden ewes were exposed to rams for 7 days (ram experienced, RE; n=61) or isolated from rams (ram na?ve, RN; n=63). All ewes were subsequently isolated from rams. In Experiment 1, RE (n=55) and RN (n=57) ewes were introduced to rams during late anoestrus. RE ewes had more total and positive interactions with rams than RN ewes (P<0.001). RE ewes showed more ram seeking behaviour and spent more time in proximity of rams than RN ewes (at least; P<0.05). In Experiment 2, RE (n=6) and RN (n=6) ewes were introduced to rams midway through a frequent blood sampling regime in late anoestrus. Ram introduction stimulated an increase in LH pulse frequency and basal LH in both RE and RN ewes (at least P<0.05). RE ewes had an increase in mean LH concentrations (P<0.01) that failed to reach significance in RN ewes (P<0.1). There was no significant effect of prior experience with rams on LH pulse frequency, amplitude or whether ewes had an LH surge. In conclusion, prior experience with rams is important in developing appropriate ewe-ram interactions but is not a pre-requisite to the endocrine response to the ram effect.     

Continuous exposure to sexually active rams extends estrous activity in ewes in spring

Sexual activity in sheep is under photoperiodic control, which is the main environmental factor responsible for the seasonality of reproduction. However, other natural environmental factors such as presence of conspecifics can slightly influence the timing of onset and offset of the breeding season. In goats, we have found that the continuous presence of bucks that were rendered sexually active out of season by previous exposure to long days, prevented goats from displaying seasonal anestrus, which suggests that the relative contribution of photoperiod in controlling seasonal anestrus should be reevaluated in small ruminant species. The aim of this study was to assess whether the presence of sexually active rams that had been stimulated by artificial photoperiod and melatonin implants, reduces seasonal anestrus in sheep, by prolonging ovulatory activity in spring. Ewes were assigned to one of two groups (n = 16 and 15), which were housed in two separate barns, and kept in contact, either with the treated or the control rams between March and July. Vasectomized rams were either exposed to 2 months of long days followed by the insertion of three subcutaneous melatonin implants (treated rams, n = 8), or exposed to natural light conditions (control rams, n = 2). Estrus was monitored daily, and weekly plasma progesterone analyses indicated ovulatory activity. Ewes that were exposed to treated rams exhibited a higher proportion of monthly estrus than ewes exposed to the control rams (P < 0.05). Thirteen of 15 ewes (one ewe was not considered because of the presence of persistent CL) exposed to stimulated rams exhibited estrous behavior in a cyclic manner. In contrast, all ewes exposed to control rams stopped estrous activity for a period of time during the study, such that this group exhibited a significantly longer anestrous season (mean ± standard error of the mean 89 ± 9 days) than did the ewes housed with treated rams (26 ± 10 days; P < 0.0001). Among 15 ewes housed with treated rams, 13 of them exhibited continuous ovulatory activity between March and July, whereas one stopped in June and two in July. All ewes kept with control rams stopped ovulating for some time; consequently, those ewes had a longer anovulation period than did the group exposed to treated rams (3 ± 3 vs. 18 ± 7 days, respectively; P < 0.05). In conclusion, continuous exposure to sexually activated rams induced by artificial photoperiod and melatonin implants in spring extended the ovarian activity of ewes in spring, which results in an increase in estrous expression.     

Effect of rumen degradable bolus containing melatonin or progestagen pessary plus pregnant mare serum gonadotropin on estrus response and lambing rate in ewes

Two practical regimens designed to induce estrus and ovulation in ewes in late anestrus were compared. Forty ewes were given a soluble glass rumen bolus containing 150 mg melatonin on July 9 and were joined with two vasectomized rams on July 23 and with three fertile rams on August 6. A second group of 40 ewes was treated with an intravaginal progestagen pessary (60mg medroxy-progesterone acetate) on July 23. Following pessary removal after 12 d, ewes were given 750 IU of pregnant mare serum gonadotropin (PMSG). Five fertile rams were joined with these ewes 48 h after progestagen removal. Melatonin concentrations were determined in single blood samples collected in early afternoon of July 21. Mating dates, lambing dates and litter sizes were recorded. Date of mating was significantly later in ewes treated with melatonin compared with those treated with progestagen plus PMSG (P<0.0001). All ewes given melatonin were mated within 4 wk, and those on progestagen plus PMSG treatment within one day of fertile ram introduction. Thirty-four ewes (85%) allocated to melatonin treatment and 36 (90%) allocated to progestagen plus PMSG treatment lambed (P>0.05). Mean (+/-SEM) lambing date was later in melatonin-treated ewes (January 17+/-1.2 d) compared to those given progestagen plus PMSG (December 30+/-0.6 d; P<0.0001). Mean litter size was lower in melatonin-treated ewes (1.5+/-0.1) compared with those given progestagen plus PMSG (2.0+/-0.1; P<0.001). Plasma melatonin concentrations indicated that 9 of 40 ewes treated with melatonin had circulating melatonin concentrations of less than 16 pg/ml. It is concluded that under conditions that existed in this experiment, treatment with progestagen plus PMSG in late anestrus resulted in more synchronous mating and lambing and a higher litter size than that following administration of a soluble glass rumen-degradable bolus containing melatonin.     

Effect of monensin and progesterone priming on ram-induced reproductive performance of boutsiko mountain breed ewes

The effects of monensin and progesterone priming on reproductive performance (estrous response, lambing rate and prolificacy) of grazing Boutsiko mountain breed adult and 18-mo.-old ewes at the end of seasonal anestrus were investigated. In Experiment 1 the feed supplement with or without monensin was offered for 21 d after introduction of vasectomized rams (Day 0). Progesterone was administered to the ewes in the respective groups as a single injection at Day -3. Ewes of both age groups were assigned randomly to 1 of 4 treatments: C, C+P, C+M and C+M+P. In Experiment 2 the supplement C or M was offered from Day -26 to Day 21. The treatments consisted of C, C+P and C+M+P. Blood samples were taken 50 h after ram introduction for determination of plasma concentrations of P and insulin-like growth factor-I (IGF-I). There was a greater increase in estrous response at Days 17 to 19 and at Days 0 to 19 when supplementation was offered before rather than after ram introduction in both age groups. In the adult group ewes synchronization of estrus at Days 17 to 19 was significantly increased by administration of monensin (P<0.05) and progesterone (P<0.01) compared with the control group in the first but not the second experiment. The incidence of estrus at Days 17 to 19 or at Days 0 to 19 was highest in the adult groups treated with monensin and progesterone in both experiments. In 18-mo.-old ewes progesterone was effective in synchronizing estrus only in Experiment 2. Mean plasma IGF-I concentrations were increased by monensin treatment (P<0.05) in adult ewes that were at the periovulatory stage at blood sampling time. Correlation coefficients between IGF-I and progesterone concentrations in monensin plus progesterone group adults were -0.715 (P<0.02) and -0.516 (P<0.01), respectively across all treatments. The results suggest that monensin and progesterone priming improved reproductive performance, and the monensin-induced increase in plasma IGF-I levels at the periovulatory stage may be causally related to the ability of ovulatory follicles to develop into functional corpora lutea (CL).     

Diurnal variation in the response of anoestrous ewes to the ram effect

The re-introduction of rams after a period of separation was used to stimulate LH secretion and induce ovulation in seasonally anovulatory ewes maintained under natural photoperiod. In 2 experiments, the rams were introduced in the morning or the evening to test for diurnal variations in responsiveness to the treatment. In the first experiment, with Romanov ewes, the ram-induced increase in tonic LH secretion was significantly earlier in the ewes treated (N = 6) at 07:30 h (mean +/- s.e.m. delay to first pulse: 20 +/- 6 min) than in those (N = 5) treated at 19:30 h (66 +/- 15 min; P = 0.006). The pulse interval after the ram effect was significantly shorter in ewes that subsequently ovulated (120 +/- 10 min) than in ewes that did not ovulate (288 +/- 108 min; P = 0.043). There was a significant decline in pulse amplitude from 6.7 +/- 1.2 to 3.4 +/- 0.6 ng/ml (both groups combined) after the introduction of rams (P = 0.040). Of the 11 ewes, 7 subsequently ovulated and a preovulatory LH surge was observed in 6 of these 30-36 h after ram introduction. In the second experiment, with seasonally anoestrous Préalpes-du-Sud ewes, the effect of the timing of the introduction of rams on the periovulatory events was tested. The delay to the onsets of oestrus and the LH surge was not affected, but the ovulation rate was higher after ram introduction in the morning (1.42) than in the evening (1.14). In the 12-h period before the introduction of the rams in the first experiment, there was a difference between the groups in the secretion of LH, but the existence of diurnal rhythms in the concentrations of LH or FSH were not confirmed in a later study in which 7 ewes were sampled every 20 min for 36 h. In contrast, there was a distinct diurnal variation in the secretion of prolactin, with the highest values being recorded at night and the lowest around midday (P = 0.025). The rise and fall in prolactin values did not appear to coincide with dawn or dusk.(ABSTRACT TRUNCATED AT 400 WORDS)