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.     

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.     

Strategies for rapid rebreeding of lactating ewes in the spring

Rapid rebreeding of winter- and spring-lambing ewes is essential if ewes are to lamb more than once per year, but fertility of lactating ewes is often low and early weaning of lambs may be undesirable in forage-based production systems. Selection to improve fertility in spring matings has been successful and led to development of ewes with a reduced seasonal anestrus. Potential for rapid rebreeding of lactating out-of-season breeding (OOS) ewes was tested in three studies. In Experiment 1, effects of short-term lamb removal on rebreeding was evaluated over 2 years using 71 January-lambing OOS ewes. At an average of 63 days postpartum, 36 ewes had lambs removed for 72 h, and all ewes were joined with rams. Circulating progesterone levels indicated that 74% of ewes ovulated before lamb separation; 91% of ewes mated within 5 weeks of ram exposure, 85% were diagnosed as pregnant and 75% lambed. The average interval between lambings was 225 days. In contrast to results observed in cattle, none of the measured variables was affected by lamb separation (P>0.20). Experiment 2 compared rebreeding performance of 24 OOS and 23 St. Croix ewes that lambed in January and averaged 60 days postpartum at ram introduction. More OOS ewes ovulated, mated and became pregnant during the first 21 days of ram exposure (83.3%, 58.3%, and 41.7%, respectively; P<0.001) compared with St. Croix ewes (26.1%, 0%, and 0%, respectively). After 39 days of ram exposure, pregnancy rates still favored OOS ewes (66.7% v. 39.1%; P=0.06), but the percentage of ewes that lambed did not differ (P>0.20) between OOS (47.8%) and St. Croix ewes (34.8%). In the third study, 34 March-lambing OOS ewes were exposed to rams on May 3 at an average of 40 days postpartum to characterize their reproductive performance. After 39 days of ram exposure, 52.9±8.7% of the ewes had mated, and 38.2±8.5% were diagnosed as pregnant. However, only 20.6±7.0% of the ewes produced viable lambs, suggesting a high level of uterine insufficiency. Spring fertility of lactating OOS ewes in these studies was one of the highest reported in the literature and indicated that selection for fertility in spring mating would improve reproductive performance in accelerated lambing programs. However, exposure of lactating OOS ewes to rams at 30 to 50 days postpartum was associated with high prenatal lamb mortality.     

Ram-induced oestrus and ovulation in lactating and weaned Corriedale ewes

Two experiments were conducted in consecutive years in which recently (Experiment 1) or temporarily (Experiment 2) weaned ewes and matched post-partum non-lactating flockmates (DRY) were exposed to a stimulus group of rams and oestrous ewes (10 and 20 in Experiment 1, 20 and 20 in Experiment 2) for 28 days in spring. Lactating ewes (n = 130) in Experiment 1 were isolated from their lambs 4 (W-4), 2 (W-2), 1 (W-1) or 0 (W-0) days in advance and exposed along with a group of 32 DRY flockmates. Lactating ewes in Experiment 2 (n = 230) were allocated to an unreplicated factorial of two levels of temporary weaning before stimulation (B0: control; B24: lambs removed 24 h before stimulation) by four levels of ewe-lamb contact imposed at the start of the stimulation (A0: control; A12, A24 and A36: lamb-ewe separation during the initial 12, 24 or 36 h of exposure); DRY ewes (n = 54) acted as an augmented factorial control. Oestrus (rump marks) and ovulation (laparoscopy on day 5 and on day 28 (Experiment 1) or day 32 (Experiment 2)) were recorded. Ovulation and oestrous responses in Experiment 1 were similar for DRY (90.6% and 55.2%, respectively) and recently weaned ewes (83.8% and 53.7%, respectively). Amongst recently weaned ewes, the immediate ovulation response to the rams and the proportion of ewes still cycling by day 28 tended to be lower (P = 0.065 and P = 0.011) in ewes weaned on the day of ram exposure (71.9% and 54.8% v. 87.8% and 80.0%, respectively). Ovulation rate was lower (P < 0.003) in W-2 ewes (1.3 ± 0.10) than in the other recently weaned groups. In Experiment 2, ovulation (83.3%) and oestrous (68.9%) responses in DRY ewes were higher (P = 0.022 and P = 0.053, respectively) than in lactating ewes (66.2% and 51.0%, respectively). More ewes ovulated (P = 0.036) in B24 (70.5%) than in B0 (61.8%). Ewes having their lambs returned 12 h after the onset of stimulation (A12) had poorer ovulation responses (54.9%) than control ewes (A0, 72.9%, P < 0.05); this was probably associated to lamb restitution after the sunset. Main conclusions were that (i) the presence of the lambs is a depressing factor of both ovulation and oestrous responses to the ram effect in lactating ewes, (ii) the ovulation response of lactating ewes will probably benefit from removing lambs for a period of 24 h before the onset of stimulation, (iii) until additional information becomes available, temporary weaning protocols should be designed avoiding lamb restitution during the night.     

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.     

Ram-induced growth of ovarian follicles and gonadotrophin inhibition in anoestrous ewes

Southdown ewes in mid-seasonal anoestrus were exposed to rams for 0 h (control group), 2 h, 24 h, 40 h, 3 days, 10 days or 20 days. Serial blood samples were then taken to determine LH and FSH levels. Ewes with greater than 24 h ram exposure were ovariectomized immediately after bleeding, and all follicles greater than 1 mm diameter were dissected from the ovaries and measured. LH basal concentrations and pulse frequency increased significantly within 2 h of ram introduction, but by 24 h fell, and then remained low. FSH concentrations fell within 2 h of ram introduction and remained low. Control group ewes (isolated) had no follicles greater than 4 mm diameter, whereas all ewes exposed to rams had large follicles, with CL or preovulatory follicles present at 40 h after ram introduction. Ram introduction was also associated with follicle recruitment (antrum formation to less than 2 mm). Follicular recruitment and development to the large follicle stage therefore occurred during a period of low plasma gonadotrophin levels and suppressed LH pulsing.     

The effects of ram exposure during progestagen oestrus synchronisation and time of ram introduction post progestagen withdrawal on fertility in ewes

Three experiments were undertaken to investigate the effect of a pre-mating ram exposure during progestagen synchronisation treatment on time of breeding, ovulation rate, embryo quality and fertility and any interaction with time of ram introduction for breeding post sponge withdrawal. Crossbred ewes in experiment 1a (n = 348), 1b (mule; n = 133) and 2 (n = 58) underwent a 12-14 days synchronisation protocol. Three days prior to sponge withdrawal ewes were divided into Control (ewes in continued isolation from rams) or +Ram (ram-exposed) groups. Rams were introduced to +Ram ewes and remained with ewes until sponge withdrawal. Ewes in experiments 1a and 2 received eCG at sponge withdrawal and were reintroduced to rams at either 36 or 48 h post sponge removal (PSR). In experiment 1b, ewes did not receive eCG and were reintroduced to rams at 24 h PSR. In experiments 1a and 1b time of breeding, date of lambing and litter size were recorded. In experiment 2, ewes were slaughtered 5 days post breeding, reproductive tracts flushed and corpora lutea, ova and embryos assessed. Fewer +Ram ewes were mated by 96 h PSR (P < 0.001) than Control ewes in experiment 1a but not when rams were introduced earlier in experiment 1b. In experiment 1a, ram introduction at 36 h PSR improved conception to first service compared to introduction at 48 h PSR (P < 0.01) in both +Ram and Control groups. In experiments 1a and 1b, +Ram ewes had reduced litter size caused by more single births (1a; P < 0.001, 1b; P < 0.01). In experiment 2, +Ram ewes had fewer corpora lutea than Control ewes (P < 0.001) but embryo quality was similar. However, more good embryos were produced when rams were introduced for breeding at 36 h compared to 48 h PSR (P < 0.001). We conclude that a pre-mating ram exposure during the synchronisation treatment reduced the number of ewes mated at and conceiving to the first service. This was partially overcome by introducing rams for breeding earlier (24 or 36 h compared to 48 h PSR) but the most dramatic decrease in fertility was due to a reduction in ovulation rate in the ram-exposed ewes.     

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.     

Combination of the ram effect with PGF2α estrous synchronization treatments in ewes during the breeding season

The role of the ram effect on the reproductive performance of ewes that have initiated estrous cycles following lambing in combination with synchronization of estrus using PGF(2α) was examined. A total of 1264 Corriedale × Merino ewes in the breeding season (March-April) were allocated to one of three treatments. The control group (PG2) of ewes (n=415) were in permanent direct contact with vasectomized rams throughout the experiment from 60 d prior to the administration of the first luteolytic dose of PGF(2α) which was followed by a second dose 13 d later (Day 0 of the experiment). Ewes assigned to the other two treatments remained isolated from rams until Day 0. In the second treatment, ewes (PG2RE; n=445) were administered PGF(2α) in the same manner and were joined with vasectomized rams at Day 0. Ewes allocated to the third treatment (PGRE; n=404) did not receive the second dose of PGF(2α) but were introduced to vasectomized rams on Day 0. Sexual receptivity, as indicated by tail-head marking, was recorded until d 11. More PG2RE ewes (407/445; 92%) were observed in estrus by Day 11 than occurred for PG2 ewes (353/415; 85%; P=0.003). The accumulated frequency of PG2RE ewes in estrus was greater than for PG2 ewes for each period from Day 3 (P<0.001) to Day 11 (P<0.01). The onset of estrus was earlier in PG2RE ewes (2.98±0.07 d) than for PG2 ewes (3.31±0.07 d; P<0.0001). In contrast, the total frequency of PGRE ewes observed in estrus by Day 11 (356/404; 88%) was similar to that observed for PG2 ewes. However, the trajectory of the accumulated frequency of the incidence of estrus was less for the PGRE ewes initially, particularly during the period of Days 3-6 of observation (P<0.0001). Consequently, onset of estrus was earlier in PG2 ewes (3.31±0.07 d) than for PGRE ewes (5.30±0.11 d; P<0.0001). It was concluded that the introduction of vasectomized rams simultaneously with the second administration of PGF(2α) advanced the onset of estrus and increased the number of ewes that responded. The introduction of rams 13 d after a single dose of PGF(2α) did not substitute for the second administration of PGF(2α).     

Supplementation of Merino ewes with vitamin E plus selenium increases α-tocopherol and selenium concentrations in plasma of the lamb but does not improve their immune function

Vitamin E and selenium have been reported to improve immune function across a range of species. Ewes lambing on poor-quality dry pasture in autumn in Western Australia are at risk of being deficient in vitamin E and selenium at lambing thus predisposing their lambs to deficiencies and increasing the risk of infection and disease. This study tested the hypotheses that (i) supplementation of autumn-lambing ewes with vitamin E plus selenium in late gestation will increase the concentrations of vitamin E and selenium in plasma in the ewe and lamb and (ii) that the increased concentrations of vitamin E and selenium in plasma in the lambs will improve their innate and adaptive immune responses and thus survival. Pregnant Merino ewes were divided into a control group (n=58) which received no supplementation or a group supplemented with vitamin E plus selenium (n=55). On days 111, 125 and 140 of pregnancy ewes in the vitamin E plus selenium group were given 4 g all-rac-α-tocopherol acetate orally. On day 111 the ewes were also given 60 mg of selenium as barium selenate by subcutaneous injection. The concentrations of α-tocopherol and selenium were measured in ewes and/or lambs from day 111 of pregnancy to 14 weeks of age±10 days (weaning). Immune function of the lamb was assessed by analysing the numbers and phagocytic capacities of monocytes and polymorphonuclear leucocytes and plasma IgG and anti-tetanus toxoid antibody concentrations between birth and 14 weeks of age±10 days. Maternal supplementation with vitamin E plus selenium increased the concentration of α-tocopherol in plasma (1.13 v. 0.67 mg/l; P<0.001) and selenium in whole blood (0.12 v. 0.07 mg/l; P<0.01) of the ewes at lambing compared with controls. Supplementation also increased the concentration of α-tocopherol (0.14 v. 0.08 mg/l; P<0.001) and selenium (0.08 v. 0.05 mg/l; P<0.01) in lambs at birth compared with controls. There was no significant effect of supplementation on immune function or survival in the lambs.     

The introduction of rams induces an increase in pulsatile LH secretion in cyclic ewes during the breeding season

Application of the ram effect during the breeding season has been previously disregarded because the ewe reproductive axis is powerfully inhibited by luteal phase progesterone concentrations. However, anovulatory ewes treated with exogenous progestagens respond to ram introduction with an increase in LH concentrations. We therefore tested whether cyclic ewes would respond to ram introduction with an increase in pulsatile LH secretion at all stages of the estrous cycle. We did two experiments using genotypes native to temperate or Mediterranean regions. In Experiment 1 (UK), 12 randomly cycling, North of England Mule ewes were introduced to rams midway through a frequent blood-sampling regime. Ewes in the early (EL; n=3) [corrected] and late luteal (LL; n=6) phase responded to ram introduction with an increase in LH pulse frequency and mean and basal concentration [corrected] of LH (at least P<0.05). In Experiment 2 (Australia), the cycles of 32 Merino ewes were synchronised using intravaginal progestagen pessaries. Pessary insertion was staggered to produce eight ewes at each stage of the estrous cycle: follicular (F), early luteal (EL), mid-luteal (ML) and late luteal (LL). In all stages of the cycle, ewes responded to ram introduction with an increase in LH pulse frequency (P<0.01); EL, ML and LL ewes also had an increase in mean LH concentration (P<0.05). In conclusion, ram introduction to cyclic ewes stimulated an increase in pulsatile LH secretion, independent of ewe genotype or stage of the estrous cycle.     

Reproductive performance of ewes exposed to vasectomized rams before breeding and forced exercise before lambing

Twenty days before a regular fall breeding season, 93 mature Rambouillet ewes were randomly allotted to one of two groups to examine the response of cycling females exposed to sterile rams. Six vasectomized rams were joined with 46 ewes during the 20-day period while 47 ewes remained separate from the rams. All ewes were judged to have been cycling either by paint marks from rams on treated animals or by the cyclic nature of progesterone (sampled at four-day intervals) profiles in control ewes. After removal of sterile males, fertile Debouillet rams remained with the ewes during a 34-day breeding season. Approximately six weeks before beginning the lambing season, one-half the females in each sterile ram treatment group were forced to walk 0.8 km per day to examine the influence of exercise on subsequent reproductive performance. Presence of sterile males did not alter (P>0.10) lambing rate, average lambing date or percentage of ewes lambing during thirds of the lambing season suggesting that prebreeding exposure of cycling ewes to vasectomized rams does not enhance lambing rate or shorten the lambing season. Forced exercise resulted in increased daily feed consumption which was reflected in heavier (P<0.05) lamb birth weights. Percentage of ewes experiencing either dystocia or pregnancy toxemia was similar (P>0.20) in exercised and unexercised animals. Moderate exercise during late gestation may increase lamb birth weight without increasing lambing difficulty.     

Heritability of ram mating success in multi-sire breeding situations

Multi-sire mating of a mob of ewes is commonly used in commercial sheep production systems. However, ram mating success (defined as the number of lambs sired by an individual) can vary between rams in the mating group. If this trait was repeatable and heritable, selection of rams capable of siring larger numbers of lambs could reduce the number of rams required for mating and ultimately lead to increased genetic gain. However, genetic correlations with other productive traits, such as growth and female fertility, could influence the potential for ram mating success to be used as a selection trait. In order to investigate this trait, parentage records (including accuracy of sire assignment) from 15 commercial ram breeding flocks of various breeds were utilised to examine the repeatability and heritability of ram mating success in multi-sire mating groups. In addition, genetic and phenotypic correlations with growth and female fertility traits were estimated using ASReml. The final model used for the ram mating success traits included age of the ram and mating group as fixed effects. Older rams (3+years old) had 15% to 20% greater mating success than younger rams (1 or 2 years of age). Increasing the stringency of the criteria for inclusion of both an individual lamb, based on accuracy of sire assignment, or a whole mating group, based on how many lambs had an assigned sire, increased repeatability and heritability estimates of the ram mating success traits examined. With the most stringent criteria employed, where assignment of sire accuracy was >0.95 and the total number of lambs in the progeny group that failed to have a sire assigned was<0.05, repeatability and heritability for log_e(number of lambs) was 0.40±0.09 and 0.26±0.12, respectively. For proportion of lambs sired, repeatability and heritability were both 0.30±0.09. The two ram mating traits (log_e(nlamb) and proportion) were highly correlated, both phenotypically and genetically (0.88±0.01 and 0.94±0.06, respectively). Both phenotypic and genetic correlations between ram mating success and growth and other female fertility traits were low and non-significant. In conclusion, there is scope to select rams capable of producing high numbers of progeny and thus increase selection pressure on rams to increase genetic gain.     

Effects of duration and timing of progesterone priming on the incidence of corpora lutea with a normal life-span in Merino ewes induced to ovulate by the introduction of rams

Three experiments investigated the effects of progesterone on the incidence of corpora lutea with normal life-span in seasonally anovulatory Merino ewes induced to ovulate by the introduction of rams.Ewes in Experiments 1 and 2 received a single i.m. injection of 20 mg progesterone in oil at the time of, or up to 5 days before introduction of rams. Control ewes received oil alone. Progesterone priming had no effect on the proportion of ewes induced to ovulate, and the beneficial effect of progesterone on the proportion of corpora lutea with a normal life-span (P < 0.001, Experiment 1; P < 0.01, Experiment 2) was independent of its time of administration. Injection of progesterone delayed the time of the surge of LH compared to control ewes (P < 0.001), but only when given at the time of introduction of rams.Administration of progesterone by intravaginal devices for 6 h, 12 h or 24 h before introduction of rams (Experiment 3) increased the incidence of corpora lutea with normal luteal life-span compared to control ewes, but had no effect on the proportion of ewes ovulating. The incidence of corpora lutea with a normal life-span was significantly and positively correlated (R=0.977; P < 0.05) with duration of progesterone priming before introduction of rams.These results indicate that duration but not timing of progesterone priming is critical to its ability to eliminate short life-span corpora lutea. Such findings support the concept of a mode of action of progesterone at the ovarian level.     

Effects of melatonin implants in ram lambs

Sixteen pinealectomized and 19 unoperated ewes were exposed to constant light for about 4 weeks before and 4 weeks after lambing. Six ram lambs born to unoperated ewes were implanted s.c. with melatonin sachets while 8 ram lambs were implanted with empty sachets. The 8 ram lambs born to pinealectomized dams also received empty implants. Ewes and lambs were then returned to the field. Analysis of weekly blood samples indicated that prolactin secretion was significantly decreased in the ram lambs with empty implants between 44-51 weeks of age whereas lambs treated with melatonin failed to show a significant change during development. All 3 groups of rams had elevated LH levels between 7 and 17 weeks of age, and a second period of high LH between 27 and 40 weeks. There were no significant differences between groups in the patterns of FSH secretion; FSH was highest between 7 and 17 weeks of age, and again between 27 and 40 weeks of age. Plasma testosterone levels in all groups increased gradually between 4 and 35 weeks. Between 38 and 48 weeks of age testosterone concentrations were markedly elevated in all groups. Growth was not affected by melatonin treatment. These results indicate that neonatal melatonin treatment has subtle endocrine effects; whether these effects are sufficient to compromise fertility in the ram, however, remains to be established.     

Ovulation rate in ewes after single oral glucogenic dosage during a ram-induced follicular phase

A 2-factor factorial array with three replicates (N = 280) was used to simultaneously assess the effects on ovulation rate of two alternative doses of medroxy-progesterone acetate (MPA) (10 or 60 mg), applied during a 6-day priming period, and the effect of a single dosage of a glucogenic formulation, administered immediately before ram exposure to groups of adult seasonally anovular Corriedale ewes. The glucogenic formulation contained 1,2,3-propanetriol (glycerol; 70% $\text{v}\text{v}$), 1,2-propanediol (propylene glycol; 20% $\text{v}\text{v}$) and distilled water (10% $\text{v}\text{v}$). At sponge withdrawal, a single oral dose of 100 ml of this formulation or the same volume of distilled water was administered to treated and control groups, respectively, and ewes were immediately exposed to rams and hormonally-induced oestrous ewes. Data from an ancillary experiment (n = 10) showed significantly (P < 0.005) above normal plasma glucose levels in treated animals at 3 and 6 h after dosage. A significant interaction (P = 0.0006) between MPA priming doses and glucogenic supplementation was detected. Supplemented ewes, among those exposed to the lower dose of MPA, exhibited a higher (P = 0.0098) mean ovulation rate (1.56 ± 0.076) than ewes that did not receive glucogenic treatment (1.31 ± 0.060). In contrast, ovulation rate was significantly decreased (P = 0.021) from 1.30 ± 0.058 to 1.13 ± 0.042 after glucogenic treatment in ewes that were primed with sponges containing 60 mg of MPA. Ewes exposed to 60 mg of MPA were marked by the rams at a significantly later (P < 0.00001) mean time (54.8 ± 1.44 h) than ewes receiving 10 mg sponges (43.6 ± 1.08 h). These results reveal the potential for modifying ovulation rate through short-term glucogenic manipulations, at least during the compressed follicular phase typical of ram-induced ovulations.     

The continuous presence of ewes in estrus in spring influences testicular volume,testicular echogenicity and testosterone concentration,but not LH pulsatility in rams

The continuous presence of active male small ruminants prevents seasonal anestrus in females, but evidence of the same mechanism operating from the females to the males is scarce. This study assessed the effects of the continuous presence of ewes in estrus in spring on ram sexual activity, testicular size and echogenicity, and LH and testosterone concentrations. On 1 March, 20 rams were assigned to two groups (n = 10 each): isolated (ISO) from other sheep, or stimulated (STI) by 12 ewes, which were separated from the rams by an openwork metal barrier, allowing contact between sexes. Each week, four ewes were induced into estrus by intravaginal sponges. Live weight, scrotal circumference, testicular width (TW) and length (TL) were recorded at the beginning and at the end of the experiment, and testicular volume (TV) was calculated; at the same time, testicular ultrasonography and color Doppler scanning were performed. Blood samples (March to May) were collected once per week for testosterone determinations, and at the end of the experiment, blood samples were collected for 6 h at 20-min intervals for LH analysis. Rams were exposed to four estrous ewes in a serving-capacity test. Scrotal circumference, TW and TL were higher in the STI than in the ISO rams (P < 0.05) in May, and TV was higher (P < 0.05) in the STI (391 ± 17 cm³) than in the ISO rams (354 ± 24 cm³). In ISO rams, the number of white pixels was higher (P < 0.01) in May (348 ± 74) than in March (94 ± 21) and differed significantly (P < 0.01) from that of the STI rams in May (160 ± 33). In ISO rams, the number of grey pixels was higher (P < 0.05) in May (107 ± 3) than it was in March (99 ± 1). Stimulated and ISO rams did not differ significantly in mean LH plasma concentrations (0.8 ± 0.5 v. 0.9 ± 0.4 ng/ml), LH pulses (2.1 ± 0.5 v. 2.2 ± 0.2) and amplitude (2.0 ± 0.4 v. 3.2 ± 0.7 ng/ml, respectively). Stimulated rams had significantly higher testosterone concentrations than ISO rams from April to the end of the experiment. Stimulated rams performed more (P < 0.05) mountings with intromission (3.0 ± 0.4) than did ISO rams (1.5 ± 0.5). In conclusion, after 3 months in the continuous presence of ewes in estrus in spring, rams had higher TV and some testicular echogenic parameters were modified than isolated rams. Although exposed rams also had higher levels of testosterone after 2 months in the presence of estrous ewes, their LH pulsatility at the end of the study was not modified.     

Developmental programming in sheep: administration of testosterone during 60-90 days of pregnancy reduces breeding success and pregnancy outcome

Evidence suggests that exposure to excess steroids during critical periods of fetal development leads to reproductive disorders. Exposure of female lambs to excess testosterone (T) from Days 60 to 90 of gestation (T60-90; term, 147 days) delayed onset of the LH surge and resulted in to male-typical reproductive behavior. The objectives of this study were to test the ability of T60-90 ewes to mate, conceive and lamb during the first three breeding seasons (Years 1, 2 and 3). Pregnant Suffolk ewes were injected with T propionate in cottonseed oil (100mg, im twice weekly) or vehicle (control; C) from Days 60 to 90 of gestation. In Year 1, ewes (C=12, T60-90=12) were kept with a vasectomized ram for 3 months and markings/visual observation of copulations were recorded. Rams had paint applied to their chest to facilitate detection of estrus and mating. All C but only three T60-90 ewes were marked (P<0.001). All ewes were then estrus-synchronized with two injections of prostaglandin F2alpha (20mg, im) given 11 days apart and allowed to mate with a painted, fertile ram. Nine of 12 C and 4 of 12 T60-90 ewes (P=0.1) were mated. Based on estrus and long-term monitoring of progesterone, more C than T60-90 became pregnant (82 and 18%, respectively; P<0.01). In Year 2, to maximize ram exposure, two C and two T60-90 estrus-synchronized ewes were placed with a painted, fertile ram at a time and mated ewes were removed to a nearby pen to force mating with others. Twenty-four hour video monitoring revealed the rams mated more C than T60-90 ewes (83 and 25%, respectively; P=0.01). In both Years 1 and 2, the rams preferred C over T60-90 ewes; therefore in Year 3 rams were given access only to T60-90 ewes. Only four T60-90 estrus-synchronized ewes were placed with a painted ram at a time. Not given an option, 91% of the T60-90 ewes were marked resulting in 4 of 11 (36%; first-service pregnancy rate in the breeding herd was 91%) ewes becoming pregnant to the synchronized estrus. Collectively these studies showed that fertility in T60-90 females was severely compromised, even after overcoming ram preference for controls.     

The Effect of oestradiol benzoate on the duration of oestrus and release of LH in ovariectomized Galway ewe lambs and adult ewes

The oestrous and LH responses by ovariectomized adult ewes (N=23) and 8-month-old ewe lambs (N=24) to i.m. injection of 10, 25, 62.5 or 156.25 μg oestradiol benzoate (ODB) were compared. The animals were primed by six daily injections of progesterone and ODB was administered 48 h after the last progesterone injection. The interval between ODB injection and onset of oestrus declined linearly (P<0.01) as the dose of ODB increased and was similar for the two age groups. The mean (±SEM) intervals to oestrus for levels of 10, 25, 62.5 and 156.25 μg ODB were 22.9±1.90, 18.0±1.33, 14.5±1.26 and 13.5±1.32 h, respectively. The duration of oestrus, determined by checking with Finnish Landrace rams at 3-h intervals, increased linearly (P<0.01) as the dose of ODB was raised and was significantly longer for ewe lambs (63.1±2.95 h) than for adult ewes (50.4±3.52 h). The overall mean (±SEM) durations of oestrus for levels of 10,25, 62.5 and 156.25 μg ODB were 16.9±5.91, 37.0±4.13, 75.2±3.94 and 97.8±4.13 h, respectively. A “pre-ovulatory” -type LH surge was observed in 32 of the 47 animals studied. The interval between injection of ODB and the beginning of the LH release declined as the dose of ODB increased (P<0.01) and was shorter (P<0.01) for ewe lambs (19.8±0.74 h) than for adult ewes (23.2±0.90 h). There was no evidence for an effect of either ewe age or dose of ODB on the maximum LH concentration observed, duration of LH discharge or total quantity of LH released. The sensitivity of the two age groups to the negative feedback effects of ODB on LH secretion was similar.     

The ovarian and hormonal response of the ewe to stimulation by the ram early in the breeding season

The introduction of Dorset rams to Romney ewes at the beginning of the breeding season (February 14 to March 1) stimulated 39% to 70% of the non-cycling ewes to ovulate. Most of the ewes that ovulated did so within 65 to 72 hours of ram introduction. The ovulations were preceded by LH peaks, the mean onset of which was 35.0±4.8 (SE) hours after ram introduction. The mean oestradiol-17β concentration per ewe ranged from 0.3 to 14.9 pg/ml plasma and there were large fluctuations among the samples collected every 3 hours. All ewes, irrespective of treatment, had similar mean concentrations of oestradiol-17β and ovarian follicular activity, and there were no changes in oestradiol-17β concentration that could be attributed to the presence of the rams.