Use of gonadotropin releasing hormone to improve reproductive responses of ewes introduced to rams during seasonal anestrus

Three experiments were conducted on anestrous ewes of Suffolk, Dorset, and Katahdin breeding to examine the potential value of GnRH to improve ovulation and pregnancy in response to introduction of rams. In Experiment 1, treatment with GnRH 2 d after treatment with progesterone (P₄; 25 mg i.m.) at introduction of rams was compared to treatment with P₄ alone at the time of introduction of rams. Treatment with GnRH did not increase percentages of ewes with a corpus luteum (CL) 14 d after introduction of rams, pregnant 32 d after treatment with PGF₂α 14 d after introduction of rams, or percent of treated ewes lambing to all services. In Experiment 2, treatments with GnRH on day 2, 7, or both after introduction of rams were compared. Treatments did not differ in mean estrous response, percentages of ewes with a detectable CL or number of CL present on day 11, or mean pregnancy and lambing rates. Therefore, neither one nor two injections of GnRH at these times appeared to be effective to induce anestrous ewes to breed. In Experiment 3, treatments compared included GnRH 4 d before introduction of rams, GnRH 4 d before and 1 d after introduction of rams, ram introduction alone, and treatment with P₄ (25 mg i.m.) at the time of introduction of rams. Percentages of ewes with concentrations of P₄ greater than 1 ng/mL (indicating formation of CL had occurred) 7 d after ram introduction tended to be greater (P < 0.07) in ewes treated with GnRH or P₄ than in control ewes treated with ram introduction alone. However, there was no difference in P₄ concentrations between groups by day 11 or 12 after introduction of rams. Estrous response rates and percentages of ewes pregnant 95 d after PGF₂α was administered (on day 12 after introduction of rams) tended to be greater (P = 0.08 and 0.06, respectively) in ewes treated with GnRH or P₄ than in ewes exposed to rams only. There was no difference in response variables between ewes treated with GnRH 4 d before introduction of rams and ewes treated with GnRH 4 d before and 1 d after introduction of rams. In conclusion, treatment with GnRH 4 d before ram introduction showed promise as an alternative to treatment with P₄ to improve the ovulatory response and reproductive performance of ewes introduced to rams during seasonal anestrus.     

Effect of small doses of naloxone on the pulsatile secretion of prolactin in the crossbreed ewe during the non-breeding season

The objective of this work was to study the effect of small doses of naloxone (Nx) on the pulsatile secretion of prolactin (Pr). For this purpose 12 crossbreed ewes were selected and allocated to three groups of four. Group 1 was treated with two injections (at 7 and 19 h) of 40 microg of GnRH. Group 2 was treated with two i.m. injections (at 7 and 19 h) of 0.5mg of naloxone. And the control group 3 was sham treated with injections of 3 ml saline. Blood samples were taken at 20 min intervals during six consecutive hours after injections. When ewes were treated at 7h no significant changes were observed in concentrations of prolactin following treatment with GnRH. Values fluctuated between 200 and 210 ng/ml. In group 2 treated with naloxone there was no change in plasma Pr concentrations during the first 100 min of sampling, however 60 min after Nx treatment Pr decreased significantly (p<0.01) and thereafter Pr plasma levels were consistently less (p<0.001) than control and GnRH treated ewes for the duration of the experiment. The response of the three groups after the second injection (19 h): After the injection of GnRH plasma Pr levels followed much the same pattern observed after the initial treatment, Pr concentrations were similar to those of control ewes. Ewes treated with a second small dose of naloxone (0.5mg i.m.) however, showed a decrease in plasma Pr 60 min after the administration of the endogenous opioid antagonist. Thereafter Nx treated ewes had lesser (p<0.001) plasma Pr levels until the termination of the experiment. It was concluded that Nx an opioid antagonist administered in small intermittent doses can alter Pr plasma concentrations in the ewe, showing that endogenous opioids are important modulators of endocrine function and that the administration of small intermittent doses of opioid antagonists produce significant endocrine changes in ewes.     

The effect of method of administration on ovarian responses of seasonally anestrous ewes administered gonadotrophin releasing hormone

Mature ewes were treated during the anestrous season with saline (I) or GnRH either intramuscularly in saline (II), subcutaneously in carboxymethylcellulose (CMC) (III) or subcutaneously in gelatin capsules (IV). Fifty μg of GnRH or 1 ml of saline were administered to 22 ewes in experiment 1. In experiments 2 and 3, forty-seven and 10 ewes received 250 μg GnRH or 1 ml of saline. Ewes were bled for progesterone determination prior to treatment and up to 12 or 13 days after treatment. In experiment 3, ovaries were observed via mid-ventral laparotomy 4 days after treatment and ovarian structures recorded. Ewes were classified into one of four progesterone response categories: cyclic, transient, prolonged or no response. The only treatment that changed the progesterone response from the saline-treated controls was GnRH in gelatin capsules. More ewes in this group were classified with a prolonged progesterone response (40%) than in the saline control group (0%). GnRH (in gelatin capsules)-treated ewes in the prolonged progesterone response category had higher concentrations of plasma progesterone than GnRH (in saline or CMC)-treated ewes with a prolonged progesterone response. For the GnRH (in gelatin capsule)-treated ewes, the prolonged progesterone response was similar to progesterone in ewes during the estrous cycle and all ewes in the prolonged progesterone category had corpora lutea (experiment 3). In summary, implanting the GnRH in gelatin capsules subcutaneously in seasonally anestrous ewes increased the ovulation response and enhanced corpus luteum function over ewes administered GnRH in saline intramuscularly.     

Fetal growth and reproductive performance in ewes administered GnRH agonist on day 12 post-mating

Reproductive performance and fetal growth was determined in GnRH (4 microg synthetic GnRH agonist, Receptal) administered (i.m.) to ewes on day 12 post-mating (n = 103) compared to control ewes (n = 97) during the breeding season. Plasma progesterone and LH concentrations were analyzed. A total of 13 ewes was slaughtered on day 45 of pregnancy (six from control, seven from GnRH treated groups). GnRH administration on day 12 post-mating increased plasma progesterone concentration (4.39+/-0.25 ng/ml) compared to control group (3.43+/-0.15 ng/ml) on days 13-15 post-mating (P < 0.01). GnRH administration also increased plasma LH concentration between 1 and 4 h after GnRH administration (P < 0.01). Pregnancy rate was higher in GnRH treated group (84%) than control (66%) group (P < 0.05). The ewes in GnRH administered group had more twins (P < 0.05) than those in control group. The ovarian weights (P < 0.05) and the number of corpora lutea (CL) (P < 0.01) were greater in ewes slaughtered on day 45 of pregnancy in GnRH treated group than those in control group. GnRH administration on day 12 post-mating did not have any effect on products of conception at day 45 of pregnancy except on crown-rump length (CRL) of fetuses and cotyledon weight. CRL of fetuses and cotyledon weight in GnRH treated group was higher than those in control group (P < 0.05). In conclusion GnRH administration improved reproductive performance of ewes when administered on day 12 post-mating probably through its beneficial effect on embryo survival by enhancing luteal function, but not through stimulating fetal growth.     

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.     

Sex behavior and hormone responses in ewes administered testosterone propionate

Two ewes were administered testosterone propionate and subsequent plasma testosterone concentrations determined and male sex behavior recorded. Initially ewes were administered 50 mg of testosterone propionate every other day for 20 days. Within 6 days following the first injection, concentrations of testosterone in plasma increased to 8.0 to 10.0 ng/ml. A 50 mg injection of testosterone propionate administered every 10 days thereafter maintained concentrations of plasma testosterone at 1.0 to 3.0 ng/ml. Sex behavior tests conducted with non-estrus and estrus ewes showed that both testosterone treated ewes developed male sex behavior similar to a ram. Ewes in estrus were mounted by testosterone treated ewes an average of 6.7 ± 1.2 times during a 10 minute test whereas none of the non-estrus ewes were ever mounted. Silastic implants containing testosterone propionate placed in the ewes 83 days following the first injection maintained concentrations of plasma testosterone at 6.0 to 8.0 ng/ml for a 20 day period. Therefore, administration of testosterone propionate to ewes effectively stimulates male sex behavior and would obviate the necessity for vasectomized rams for estrus detection.     

The effects of GnRH analogue (buserelin) or hCG (Chorulon) on Day 12 of pregnancy on ovarian function, plasma hormone concentrations, conceptus growth and placentation in ewes and ewe lambs

The objectives of this study were to determine the effect of GnRH analogue (buserelin) or human chorionic gonadotrophin (hCG, Chorulon) treatment on Day 12 of pregnancy on ovarian function, plasma hormone concentrations, conceptus growth and placentation in ewes and ewe lambs. After oestrus synchronization with progestagen sponges and eCG, all the animals were mated with fertile rams. Both ewes and ewe lambs (20 per treatment group) were given either normal saline or 4 microg GnRH or 200 IU hCG on Day 12 post-mating. Pre- and post-treatment plasma hormone concentrations were determined in seven pregnant animals per treatment group in samples collected 1h before and 0, 2, 4, 6, 8, 24, 48 and 72 h after treatment. Overall mean progesterone concentrations were higher (P<0.001) in ewes as compared with ewe lambs in saline-treated controls. GnRH or hCG treatment increased (P<0.001) mean plasma progesterone concentrations in both age groups, however, post-treatment concentrations were significantly (P<0.05) higher in ewes than in ewe lambs. Oestradiol concentrations were similar in the two control groups. In ewes, but not in ewe lambs, both GnRH and hCG treatments significantly (P<0.05) increased the mean oestradiol concentrations above pre-treatment levels. Moreover, post-treatment oestradiol concentrations in GnRH- and hCG-treated animals were significantly (P<0.05) higher than those in the saline-treated controls. LH release in response to GnRH treatment was greater (P<0.05) in ewes than in ewe lambs, whereas FSH release in ewes was less (P<0.05) than that of ewe lambs. The effects of GnRH or hCG on conceptus growth and placentation was determined at slaughter on Day 25. In ewes, GnRH treatment increased (P<0.05) luteal weight, amniotic sac width and length, and crown-rump length compared with controls, but had no effect on these parameters in ewe lambs. In ewes, hCG treatment also enhanced (P<0.05) luteal weight, amniotic sac width and length, crown-rump length, embryo weight and number of placentomes as compared with controls. In ewe lambs, there was no difference (P<0.05) between hCG and control groups in luteal weight, embryo weight and amniotic sac width but crown-rump length, amniotic sac length and the number of placentomes forming the placenta were greater (P<0.05). In conclusion, GnRH or hCG treatment on Day 12 of pregnancy can increase ovarian function, conceptus growth and placental attachment in ewes. However, these treatments were less effective in ewe lambs.     

Treatment of seasonally anestrous Romney ewes with continuous infusion of low doses of GnRH: effects on estrus, ovulation and plasma progesterone concentration

The aim of this study was to assess the suitability of a GnRH infusion regimen (125 ng/h or 250 ng/h) to induce estrous behaviour, ovulation and normal corpus luteum function in progesterone-primed Romney ewes each month of seasonal anestrus (i.e. September to February inclusive) over two years. None of the progesterone-primed control ewes (i.e. no GnRH treatment; N = 120 observations) ovulated, showed normal corpus luteum function or displayed estrous behaviour at any time during anestrus. Approximately 27 and 50% of the respective 125 ng/h and 250 ng/h GnRH-treated ewes (N = 120 observations per GnRH treatment) ovulated and showed normal luteal function. Of those which ovulated 59.2% and 52.4% in the respective 125 ng/h and 250 ng/h GnRH treatment groups showed estrous behaviour. There was a significant effect of GnRH dose on the median number of ovulations (250 ng/h > 125 ng/h; P<0.01) but no overall difference (when both treatment years and GnRH doses were pooled) in the median number of ovulations per month of anestrus. The frequency of ewes with an ovulation rate >2 was low with only 4/95 treated ewes with more than 2 corpora lutea (CL). Treatment of progesterone-primed ewes with 250 ng/h GnRH increased plasma LH (P<0.01) but not FSH concentrations; a significant increase in LH pulse amplitude (P<0.05) but not LH pulse frequency was observed. The plasma gonadotropin levels in the 125 ng/h GnRH treatment groups were not studied. We suggest that in breeds such as the Romney which have a strict (i.e. 5-6 month) anovulatory interval, the GnRH-infusion technique may be of limited practical use for inducing pregnancies during the non-breeding season.     

Pituitary receptors for gonadotropin-releasing hormone in relation to changes in pituitary and plasma luteinizing hormone in ovariectomized-hypothalamo pituitary disconnected ewes. I. Effect of changing frequency of gonadotropin-releasing hormone pulses

Studies were undertaken to determine if changes in the amplitude of luteinizing hormone (LH) pulses that occur in response to changes in the frequency of gonadotropin-releasing hormone (GnRH) pulses are due to an alteration in the number of GnRH receptors. Ewes were ovariectomized (OVX) and the hypothalamus was disconnected from the pituitary (HPD). Ewes were then given pulses of GnRH at a frequency of 1/h or 1/3 h. Two control groups were included: OVX ewes not subjected to HPD, and HPD ewes that were not OVX. At the end of one week of treatment, blood samples were collected to determine the amplitude of LH pulses. The treated ewes were killed just before the next scheduled pulse of GnRH, and the content of LH and number of GnRH receptors were measured in each pituitary. The amplitude of LH pulses was highly correlated with the amount of LH in the pituitary gland (r = 0.71, p less than 0.01), and both LH content and pulse amplitude (mean + SEM) were higher in ewes receiving GnRH once per 3 h (189.7 +/- 39.3 microgram/pituitary, 10.3 +/- 1.1 ng/ml, respectively) than in ewes receiving GnRH once per h (77.8 +/- 11.4 microgram/pituitary, 5.2 +/- 1.3 ng/ml). The pituitary content of LH was highest in the OVX ewes (260.2 +/- 57.4 micrograms/pituitary) and lowest in the nonpulsed HPD ewes (61.7 +/- 51.2 micrograms/pituitary). The number of GnRH receptors was similar in all groups, and was not correlated with any other variable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of a single injection of hCG or GnRH agonist on day 12 post mating on fetal growth and reproductive performance of sheep

Reproductive performance and fetal growth were determined when hCG (150 i.u. Pregnyl; n=44), GnRH (4 microg synthetic GnRH agonist, buserelin, Receptal; n=43) or saline (control, n=45) was administered (i.m.) to ewes on day 12 post mating during the breeding season. A total of 12 ewes was slaughtered on day 45 of pregnancy (four from each treatment group). Non-return rate and lambing rate were higher for ewes in the hCG (0.89 and 84%) and GnRH treated groups (0.86 and 79%) than for ewes in the control (0.69 and 62%) group (P<0.05). The ewes in the hCG and GnRH groups also had more twins (P<0.05). Birth weights of these twin lambs in the hCG and GnRH groups were heavier than those in the control group (P<0.05), but this difference had disappeared at weaning 60 days later. Lamb mortality was similar among treatment groups resulting in a higher number of lambs weaned in the hCG and GnRH groups. The ovarian weights and the number of corpora lutea (CL) and luteal weights of ewes slaughtered on day 45 of pregnancy were greater (P<0.05) in the hCG and GnRH treated groups than those measured in the control group. Administration of hCG on day 12 post mating increased gravid uterus weight, crown-rump-length (CRL), chorioallantois weight and total cotyledon weight (P<0.05) of conceptuses recovered on day 45 of pregnancy compared to the control group. The weights of caruncules (P<0.05) and placenta (P<0.01) were higher in conceptuses of both the hCG and GnRH groups. The weights of fetuses in the hCG group were higher than those in both the GnRH and control groups (P<0.05). In conclusion, both hCG and GnRH administration improved reproductive performance of ewes when administered on day 12 post mating. However, hCG and GnRH appeared to act differently on embryo survival because only hCG administration increased fetal growth.     

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.     

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.     

Effect of hysterectomy on the short life-cycle corpus luteum produced after GnRH-induced ovulation in the anoestrous ewe

Anoestrous Romney Marsh ewes were treated with small-dose (250 ng) multiple injections of GnRH. Ewes in Groups 1 and 3 were hysterectomized 2 weeks before treatment, while those in Groups 2 and 4 were intact controls. Groups 1 and 2 were primed with progesterone (+P) and treated with 2 h injections of GnRH (250 ng) for 36 h, while Groups 3 and 4 were not pretreated (-P) but were given 2 h injections of GnRH (250 ng) for 18 h. Both treatment regimens were terminated with a bolus injection of GnRH (125 micrograms), given to synchronize the timing of the LH surge and subsequent luteal progesterone production. The plasma progesterone profiles of 5/5 animals in Group 2 (+P controls) and 2/5 animals in Group 4 (-P controls) were indicative of normal luteal function, while the remaining 3/5 animals in Group 4 produced plasma progesterone profiles typical of abnormal luteal function. However, in all the hysterectomized animals (Groups 1 and 3) peripheral plasma progesterone concentrations rose to reach a mean peak value of 1.3 ng/ml plasma on Day 8 which was maintained in all animals irrespective of progesterone pretreatment. The absence of a fall in progesterone concentrations precluded the identification of any animal in Group 4 showing abnormal luteal function. It was also noted that, after hysterectomy, although the corpus luteum was maintained, it was with reduced secretory capacity. The prevention of the expected proportion (70%) of -P animals from displaying a decline in plasma progesterone concentration after hysterectomy provides firm evidence that the uterus is involved in the premature regression of the short-cycle corpus luteum.     

Reproductive potential and endocrinological responses of sheep kept under controlled lighting. II. Pituitary and gonadal responses of ewes and rams to a six-monthly light cycle

Forty entire ewes of mixed breeds were kept in environmentally-controlled rooms with a 6-monthly light cycle. Six mature spayed Border Leicester × Merino ewes and four mature intact Poll Dorset rams were kept under the same conditions.Over a period of 2 years (four light cycles) estimates were made of ovarian, testicular and pituitary activity in response to the artificial light regime. Non-pregnant ewes were bled twice weekly: peripheral plasma progesterone levels > 1 ng/ml were taken as indicative of ovarian activity. Testicular activity was estimated by weekly tests for peripheral plasma testosterone and scrotal sac volume. Pituitary activity was estimated monthly by the response to the injection (i.v.) of 75 μg gonadotrophin releasing hormone (GnRH) to rams and of 18.75, 37.5 or 75 μg to ewes, using peripheral plasma luteinising hormone (LH) estimations from the time of GnRH injection.Data for ovarian and testicular measurements were classed into categories according to week and for pituitary response to month of the light cycle.Cubic regression analyses were conducted on the percentage of ewes with progesterone levels > 1 ng/ml and on ram testosterone and scrotal measurements.Despite the irregularity of the curve for the light cycle, the ovarian and testicular responses of rams and ewes followed an alternating curve with peaks and troughs of activity separated by approximately 13 weeks in the 26-week cycle. The peak of ovarian activity occurred during the long daylength period which followed a 22-week period of decreasing daylength and was preceded by 1 month by peak ram testosterone and scrotal sac volume.The pattern of pituitary response was related to that of the actual light cycle and the data were subjected to time-lag regression. This econometrical technique revealed that there was a delay in pituitary response to daylength changes of 2 months for rams, and between 2 and 3 months for the spayed ewes. The peak pituitary response to the GnRH test occurred one month earlier for rams than for the spayed ewes, and coincided with the corresponding troughs of gonadal activity of each sex.The results showed that the breeding season of sheep can be compressed into 6 months and that the pattern of pituitary response follows the daylength pattern more closely than do measurements of gonadal activity. Peak reproductive activity in rams, as measured by pituitary and gonadal activity, precedes that of ewes by approximately 1 month.     

The effect of late pregnancy supplementation of ewes with vitamin E on lamb vigour

The experiment measured lamb responses to supplementation of the pregnant ewe diet with vitamin E above requirement. Crossbred ewes were mated with either Suffolk or Texel rams. Twin-bearing ewes were randomly allocated (approximately 21 months of age at allocation) to one of four treatment groups (20 ewes per group, 10 mated with Suffolk and 10 with Texel rams). Treatments imposed were 50, 100, 150 or 250 IU supplementary vitamin E per ewe per day to give a four treatment by two sire-type factorial experimental design. Ewes were fed concentrates to meet energy requirements for stage of pregnancy and hay ad libitum. Diets were introduced approximately 6 weeks before lambing. Blood samples were obtained prior to introduction of diets, 17 days after introduction of diets and within 24 h of lambing from a subset of eight ewes per treatment (32 total). Colostrum samples were obtained from 10 ewes per treatment, 12 h after birth of the first lamb. All births were observed and a lamb vigour score was assigned to each lamb 5 min after birth. At 1 and 12 h after birth, rectal temperature, and at 12 h after birth, sex, crown-rump length and BW of each lamb were recorded. Mean ewe plasma α-tocopherol concentration prior to introduction of the diets was 1.5 μg/ml (s.e.m. 0.09) and did not differ between groups. There were positive linear (P < 0.001) effects of dietary vitamin E on plasma (17 days after introduction of diets) and colostrum (12 h after birth) α-tocopherol concentrations. Lamb vigour scores were superior (P < 0.001) for lambs sired by Texel rather than Suffolk rams but there were no differences as a result of vitamin E supplementation. Lamb mortality was low and unrelated to either sire or supplementary vitamin E. Lamb birth and weaning weights were also unaffected by vitamin E supplementation. Supplementing the ewe with vitamin E therefore had no effect on any lamb measurements.     

Estrogen and progesterone receptor content in the pituitary gland and uterus of progesterone-primed and gonadotropin releasing hormone-treated anestrous ewes

The objective of this work was to investigate the effect of progesterone (P) and gonadotropin-releasing hormone (GnRH) treatment on estrogen receptor (ER) and P receptor (PR) concentrations in the pituitary gland and uterus of anestrous ewes. Ewes were either not treated (group C, n = 4); were treated with 0.33 g P-controlled internal drug release (P-CIDR) for 10 days (group P, n = 4), with GnRH, 6.7 ng i.v. injections every 2 h for 18 h followed by a 4 microg bolus administration of Receptal at 20 h (group GnRH, n = 4), or with a combination of the P and GnRH treatment (group P + GnRH, n = 3). Ewes were humanely killed either at the beginning of the experiment (group C), when the CIDR was removed (group P), or 24 h after the GnRH bolus treatment (groups GnRH and P + GnRH). Progesterone treatment increased serum P concentrations, indicating that the treatment was effective. All GnRH treated ewes had similar luteinizing hormone (LH) surges, which lasted 8 h. At slaughter, estradiol (E2) concentrations in the GnRH group were higher than in groups C, P, and P + GnRH. Treatment with GnRH increased more than 10-fold the content of ER and PR in the pituitary gland without altering steroid receptor concentrations in the uterus. When GnRH was combined with P the uterine receptor contents were higher than with P treatment alone. The treatment with P decreased ER and PR content in the uterus, but had no effect on the pituitary gland. The results show that regulation by P and GnRH of ER and PR content in anestrous ewes is tissue-specific.     

Pulsatile GnRH administration stimulates the number of pituitary GnRH receptors in seasonally anoestrous ewes

Twenty seasonally anoestrous ewes were pretreated with progesterone for 4 days and divided into four equal groups. Ewes in Group 1 received no GnRH treatment and were slaughtered immediately after progesterone removal. Ewes in Groups 2, 3 and 4 received i.v. injections of 250 ng GnRH every 2 h for 36 h starting at the time of progesterone removal. Ewes in Group 2 were slaughtered immediately after the 36 h GnRH pulsing, while ewes in Groups 3 and 4 were given a bolus injection of 125 micrograms GnRH at this time and were slaughtered 2 and 10 h after the bolus injection, respectively. Blood samples were collected every 30 min from ewes in Group 4 only, from 4 h before the start of GnRH treatment until 10 h after the bolus injection. Pulsing with GnRH resulted in episodic release of LH, and the bolus injection of GnRH was immediately followed by a preovulatory type LH surge in those ewes in which an endogenous surge had no already begun. The pituitary GnRH receptor numbers were significantly higher for the ewes in Group 2 than for any of the other treatment groups, while there was no significant difference in the receptor numbers between Groups 1, 3 and 4. The results suggest an up-regulation of GnRH receptors resulting from pulsatile GnRH therapy.     

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.     

A new approach to enhance reproductive performance in sheep using royal jelly in comparison with equine chorionic gonadotropin

The objective was to compare the effects of royal jelly (RJ) and eCG treatments on reproductive performance of ewes synchronized using intravaginal progesterone-releasing devices. Forty-two cycling Awassi ewes were treated intramuscularly (i.m.) with 15 mg PGF2alpha. On the following day, all ewes were administered with CIDR-G for 12 days and were randomly allocated to three (RJ, eCG and control) groups of 14 ewes each. Ewes in the RJ-treated group received daily i.m. treatments of 400mg RJ during the period of CIDR-treatment. Each ewe in the eCG-treated group received an i.m. treatment of 500 IU eCG at the time of CIDR-G removal (day 0) and no further treatment was given to ewes in the control group. Ewes were exposed to four fertile rams for 72 h, from the time of CIDR-G removal, and checked for breeding marks at 6-h intervals. Blood samples were collected from day -13 until day 0 and thereafter until day 19 for progesterone analysis. Royal jelly treatment resulted in a greater rate of decline and lower (P<0.02) progesterone concentrations between days -10 and 0 than eCG-treated and control ewes. Expression of estrus was similar among the three groups and intervals to onset of estrus were shorter (P<0.01) in RJ-treated (31.3h) and eCG-treated (29.8h) than control (41.3h) ewes. First-cycle pregnancy and lambing rates were greater (P<0.05) in RJ-treated (71.4 and 71.4%) and eCG-treated (85.7 and 78.6%) than in control (42.9 and 35.7%) ewes, respectively. Results demonstrate that the treatments of RJ and eCG in conjunction with CIDR-G were similarly effective in induction of estrus and improvement of pregnancy and lambing rates.     

Effect of post-mating GnRH analogue (buserelin) treatment on PGF2alpha release in ewes and ewe lambs

The objectives of this study were to determine the effects of buserelin or saline treatment on ovarian function (Experiment 1), plasma PGFM concentrations and oxytocin stimulated prostaglandin F(2alpha) (PGF(2alpha)) release (Experiment 2) in ewe lambs and ewes. Welsh Halfbred ewes (n=26) and ewe lambs (n=24) were mated to vasectomised rams at synchronised oestrus and on Day 12 post-mating each animal was injected intramuscularly either normal saline or 4 microg buserelin. In Experiment 1, plasma progesterone and oestradiol concentrations were determined in samples collected by jugular venepuncture 1h before and at 0, 2, 4, 6, 8, 24, 48 and 72 h after treatment (n=7 per treatment group). Progesterone concentrations increased (P<0.05) from 2 to 8h after buserelin treatment and returned to basal levels after 72 h, whereas oestradiol concentrations were maximal at 2h post-treatment and returned to basal levels after 24h (P<0.05). Oestradiol concentrations were lower (P<0.05) in buserelin-treated animals than controls at 72 h post-treatment. Basal and post-treatment progesterone concentrations were greater (P<0.05) in ewes than in ewe lambs but oestradiol levels were similar for both age groups. Ovulation rate, determined by laparoscopy on Day 14, was similar for both age groups (ewes 1.1; ewe lambs 1.0). Buserelin treatment induced accessory corpora lutea in ewes (4/7; 57%) but not in ewe lambs (0/7; 0%). In the Experiment 2, plasma PGFM concentrations were determined in samples collected at 20-min intervals for 6h on Day 14 and at 20-min intervals for 1h before and at 10-min intervals for 1h and then at 20-min intervals for a further 3h period after an intravenous injection of oxytocin (1IU/kg body weight) on Day 15 post-oestrus. In this experiment there were five ewe lambs and six ewes per treatment group. There was no effect of buserelin treatment or age on basal PGFM concentrations on either Day 14 or 15. Although peak PGFM concentrations tended to be lower in buserelin-treated animals, the difference was not significant (P>0.05). However, peak duration following oxytocin challenge on Day 15 post-mating was shorter (P<0.05) in control ewes compared with control ewe lambs. In conclusion, buserelin treatment given on Day 12 post-oestrus enhances luteal function more in ewes than ewe lambs and after a transitory increase, reduces oestradiol concentrations in both ewes and ewe lambs. However, buserelin treatment does not significantly attenuate the luteolytic signal.