hCG treatment on day of mating improves embryo viability and fertility in ewe lambs

An investigation was conducted to determine the effects of supplementing hCG at mating time on luteal function, conceptus growth, placentation and reproductive performance in TexelxClun Forest ewe lambs. After oestrus synchronisation with progestagen sponges and PMSG ewe lambs were treated either with normal saline (n=102) or 150 IU of hCG (n=105) at mating time. At 25 days after mating 24 animals were slaughtered from each group to determine embryo viability and placental development. hCG significantly (P<0.05) increased crown-rump length (saline: 11.9+/-0.2 mm; hCG: 12.7+/-0.2 mm), amniotic sac width (saline: 11.4+/-0.4 mm; hCG: 12.0+/-0.3 mm) and the number of placentomes (saline: 90.8+/-7.3; hCG=122.4+/-6.3). Among the pregnant animals that were slaughtered on 25 days post-mating, ovulation rate tended to be higher in the hCG group compared to controls (saline: 1.16; hCG: 1.54). However, no difference was observed either in mean plasma progesterone concentrations (saline: 4.6 ng/ml; hCG: 4.9 ng/ml; sed 0.56) or in progesterone production from luteal slices when cultured in vitro (saline: 239.6+/-11.8 ng/mg; hCG: 263.2+/-13.6 ng/mg) between controls and hCG treated animals. Reproductive performance was also recorded in ewe lambs that were either treated with saline (n=78) or hCG (n=81). The total number of lambs born (saline: 38; hCG: 58) was significantly (P<0.05) greater in hCG group compared to saline-treated controls. Both lambing percentage (saline: 36%; hCG: 48%) and litter size (saline: 1.35; hCG: 1.48) tended to be greater (P<0.10) in hCG-treated animals compared to the controls. In conclusion, these data suggest that treatment of ewe lambs with hCG at the time of mating improves conceptus growth, placentation and number of lambs born.     

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.     

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.     

Effects of high ambient temperature on respiration rate, rectal temperature, fetal development and thyrold gland activity in tropical and temperate breeds of sheep

The effects of high ambient temperatures on rectal temperature (RT), respiration rate (RR), fetal development and serum thyroxine (T(4)) concentrations were stuaied in two experiments involving 35 ewes and 26 lambs from the following ewe groups: 1) Barbados Blackbelly (B), a tropical breed; 2) Dorset (D), a temperate breed; and 3) Blackbelly x Dorset crosses (BxD). Data were obtained on four B, five D and five BxD ewes exhibiting estrus during the summer (Exp. 1). In Exp. 2, eight B, seven D and six BxD ewes were maintained in two environmental chambers (cool, 22.2C; hot, 33.8C) from day 125 of gestation to seven days before the expected lambing date for each breed group (D and BxD, 140+/-1; B, 144+/-1 day of gestation). The B and BxD ewes were more heat-tolerant than D ewes as measured by significantly lower RT and RR in each experiment. Mean lamb birth weight, crown-rump length, number of functional uterine caruncles and caruncle weight and size did not vary significantly among breed groups or temperature chamber (Exp. 2), and there was no indication that the high temperature imposed caused fetal dwarfing in lambs removed from the uterus at a standard age of seven days before expected parturition. Serum T(4) varied markedly among breed groups (P<0.05) in each experiment with B ewes having the lowest and BxD ewes the highest concentration. In Exp. 1, follicular stage T(4) concentrations in B and BxD ewes were lower (P<0.02) than those during the luteal stage of the estrous cycle. The decrease in D ewes was not significant. High ambient temperature (Exp. 2) depressed T(4) levels in D ewes (P<0.05) and also depressed the pituitary-thyroid response to thyrotropin releasing hormone in D lambs. Such was not the case in B and BxD ewes and their lambs.     

The effect of hCG treatment on Day 12 post-mating on ovarian function and reproductive performance of ewes and ewe lambs

The objectives of this study were to determine the effect of a single intramuscular injection of 200 IU hCG (Chorulon) on Day 12 post-mating on ovarian function and subsequent lambing performance in ewes and ewe lambs bred at synchronised oestrus during the breeding season and on the lambing performance of ewes induced to breed during late anoestrus. All animals were mated to rams at synchronised oestrus and on Day 12 post-mating given normal saline or 200 IU hCG.In Experiment 1, laparoscopic results showed that hCG treatment induced accessory corpora lutea in ewes (control = 0/7; hCG = 5/7) but not in ewe lambs (control = 0/7; hCG = 0/7).In Experiment 2, hCG treatment did not improve the lambing rate (control = 50; hCG = 57) or the litter size (control = 1.80; hCG = 1.96) in ewes (control = 100; hCG = 91). However, hCG treatment significantly (P > 0.05) improved the lambing rate (control = 29; hCG = 58; P < 0.05) in ewes conceiving at the first oestrus after treatment. hCG treatment (control = 42; hCG = 42) also failed to improve the lambing rate in ewe lambs (control = 48; hCG = 41).In Experiment 3, hCG treatment had no significant (P > 0.05) effect on the lambing rate (control = 72; hCG = 62) or the litter size (control = 1.59; hCG = 1.58) in ewes (control = 111; hCG = 115) induced to breed during anoestrus or on ewes returning to oestrus and conceiving after treatment (lambing rate: control = 86; hCG = 72; litter size: control = 1.44; hCG = 1.35). In conclusion, the data obtained in this study suggest that during the breeding season hCG may, by stimulating ovarian function, improve embryo survival in ewes conceiving at the first post-treatment oestrus. This effect, however was not observed in ewe lambs.     

Effect of diet and GnRH administration on post-partum ovarian cyclicity in autumn-lambing ewes

Using autumn-lambing ewes, this study investigated (i) the effects of diet on gonadotrophin secretion and responsiveness of the hypothalamic-pituitary-ovarian axis to exogenous GnRH during the early post-partum period; and (ii) whether ovulation prior to completion of uterine involution results in an increased incidence of aberrant ovarian cycles. Thirty-two ewes rearing 1.9+/-0.12 lambs were equally allocated to two dietary treatments at lambing (22 October +/-0.2 day). Diets comprised ad libitum hay and 1.5 kg per ewe per day of one of two concentrates (11.5 MJ ME, 195 g CP per kg) containing 300 g kg(-1) cracked maize grain (M) or 300 g kg(-1) sugar beet pellets (S). Half of the ewes on each diet (G) received 25 i.v. injections of 250 ng GnRH in 2 ml 0.9% saline at 2 h intervals from days 12-14 post-partum while remaining ewes (N) were monitored for the resumption of spontaneous ovarian cyclicity. Blood samples were obtained from all ewes throughout the study (lambing to 18 December) for measurement of circulating hormone concentrations and the uteri and ovaries of all ewes were examined via laparoscopy on day 21 post-partum. There were no effects of dietary treatment on ewe daily live weight loss, lamb daily live weight gain or the immediate post-partum increase in circulating FSH concentrations. Diet did not affect insulin concentrations or LH pulse frequency on day 12 post-partum but LH pulse amplitude was lower in ewes fed concentrate M compared to concentrate S (1.4+/-0.10 versus 1.7+/-0.12 ng ml(-1), respectively, P<0.05) and this was associated with an increased interval to the resumption of spontaneous ovarian cycles (35+/-3.1 versus 26+/-2.1 day, respectively, P<0.05). Administration of exogenous GnRH increased (P<0.05) the proportion of ewes on both diets that ovulated within 20 days of parturition and advanced the onset of ovarian cyclicity in ewes fed concentrate M by 9.5 days (significance of interaction, P<0.05). Four ewes, all of which ovulated before day 22 post-partum, had extended luteal activity while in remaining ewes, duration of the first luteal phase was inversely related to the time of first ovulation (r(2)=0.16, P<0.05). Results demonstrate that (i) the onset of ovarian cyclicity is influenced by diet and can be advanced by administration of exogenous GnRH; and (ii) ovulation during the early post-partum period is associated with an increased incidence of extended luteal activity.     

Luteal function and blastocyst development in ewes following treatment with PGF(2)alpha and GnRH

Luteal function and blastocyst development were compared in ewes treated with GnRH (100 mug) on Day 1 (Day 0 = day of estrus) or in ewes previously induced into estrus with PGF(2)alpha. In Experiment 1, the duration of estrous cycles of ewes previously treated with PGF(2)alpha were longer (P<0.06) than those that received PGF(2)alpha plus GnRH, GnRH alone, or remained untreated (control) ewes. Progesterone concentrations were lower (P<0.07) on Day 1 and higher (P<0.01) on Days 16 and 17 of the estrous cycles following PGF(2)alpha treatment relative to those of the natural (control) cycles. In Experiment 2, blastocysts of ewes treated with PGF(2)alpha were less developed (P<0.06) by Day 13 of pregnancy than those of the control ewes. The GnRH treatment did not influence any of these characteristics. Treatment with PGF(2)alpha delayed luteal formation during the subsequent estrous cycle, increased the duration of the estrous cycle and slowed the rate of blastocyst development relative to GnRH-treated and untreated ewes.     

Function of abnormal corpora lutea in vitro after GnRH-induced ovulation in the anoestrous ewe

Normal and abnormal corpora lutea were recovered from anoestrous Romney Marsh ewes on Days 3, 4, 5 and 6 after treatment with small-dose (250 ng) multiple injections of GnRH followed by a bolus injection (125 micrograms) with (+P) and without (-P) progesterone pretreatment and a study made of their characteristics in vitro. Plasma progesterone concentrations initially rose concurrently in all animals but abnormal luteal function occurred in 70% of the -P ewes and was defined on Day 5 when plasma progesterone concentrations declined relative to those in the +P ewes. All corpora lutea recovered on Days 3 and 4 appeared macroscopically similar and there were no significant differences between the +P and -P groups in terms of luteal weight, progesterone content and binding of 125I-labelled hCG on these days. However, corpora lutea from the -P animals only exhibited a decline in progesterone production in vitro on Day 4 (P less than 0.01), and morphological differences became apparent on Days 5 and 6 when the abnormal corpora lutea from the -P animals also decreased in weight (P less than 0.01) and progesterone content (P less than 0.001). Binding of 125I-labelled hCG increased on Day 5 in the normal corpora lutea only. These results show that, although abnormal luteal function induced by GnRH treatment of anoestrous ewes could not be distinguished from normal corpora lutea before Day 5 by measurement of progesterone in peripheral plasma, a significant decline in progesterone production in vitro occurred on Day 4 in the abnormal corpora lutea. This was followed by significant decreases in weight and progesterone content and a failure to increase 125I-labelled hCG binding. Abnormal corpora lutea are therefore capable of some initial growth and progesterone production, before undergoing a rapid and premature regression from Day 4, which has similar characteristics to natural luteolysis.     

Effects of exogenous progesterone on gestation length, foetal survival and colostrum yield in ewes

Twin bearing mature ewes (n=40) were treated with exogenous progesterone (100mg daily in oil) or vehicle (oil control) from Day 143 of gestation until lambing to investigate the effects on gestation length, foetal survival and colostrum yield and composition. Compared to control ewes, progesterone treated ewes had increased (P<0.05) serum progesterone concentrations (by 4.3 ng/ml) before lambing and in the first day post-partum (by 10 ng/ml). Progesterone treatment increased gestation length (150.4+/-0.6 days versus 147.8+/-0.6 days, P<0.05) and colostrum yield at 1h after lambing (P<0.05) but the colostrum had a lower concentration of IgG (P=0.02). In the first 24h after lambing, total colostrum and IgG yields were not different between groups. Four (20%) of the progesterone treated ewes produced either one or two dead lambs, while one ewe died on day 155 without initiating the birth process. We conclude that the daily administration of 100mg progesterone resulted in extended gestation length and reduced lamb survival but did not lower colostrum yield.     

Effects of hormonal manipulation on the resumption of postpartum reproductive activity in Texel ewes

Three experiments were conducted on Texel ewes to study the influence of prostaglandin F(2alpha) (PGF(2alpha)), prolactin (PRL), estradiol (E(2)), and gonadotrophin releasing hormone (GnRH) on postpartum reproductive activity. In Experiment 1, oral administration of indomethacin (25 to 50 mg/day/ewe) from Day 3 post partum to the first detected estrus inhibited plasma 13, 14-dihydro-15-keto, PGF(2alpha) (PGFM) concentrations (P < 0.0001). This treatment resulted in an earlier rise in the frequency and amplitude of luteinizing hormone (LH) pulses and a resumption of estrous behavior (P < 0.05), while ovarian activity estimated by progesterone (P(4)) concentrations resumed to the same extent in treated ewes and controls. Bromocriptine treatment (2.5 mg/day/ewe) reduced plasma PRL levels (P < 0.0001) but had no effect on ovarian activity as evidenced by P(4) and resumption of estrus or on either the frequency or amplitude of the LH pulse. In Experiment 2, a single injection of GnRH agonist (42 mcg of buserelin/ewe) on Day 16 post partum resulted in an abrupt elevation of plasma LH concentrations; mean LH values were 18 to 27 times higher when compared with those of the control ewes. Two days after this treatment, ovulations occurred in 5 of the treated ewes and in 2 of the control ewes. This induced ovarian activity was not associated with estrous behavior; however, after an adequate subsequent luteal phase all the treated ewes displayed estrus, the resumption of estrus thus being earlier in treated than in control ewes (P < 0.01). In Experiment 3, E(2) supplementation from Day 16 to Day 28 post partum increased the number of LH pulses per 6 hours in suckling ewes (P < 0.05) and induced earlier resumption of estrus in dry ewes but not in suckling ewes (P < 0.01). Luteal function was detected about 5 and 8 days after the insertion of E(2) implants in 4 dry ewes and in 2 suckling ewes, respectively.     

Ovarian function in Nelore (Bos taurus indicus) cows after post-ovulation hormonal treatments

Maternal recognition of pregnancy in the cow requires successful signaling by the conceptus to block luteolysis. Conceptus growth and function depend on an optimal uterine environment, regulated by luteal progesterone. The objective of this study was to test strategies to optimize luteal function, as well as prevent a dominant follicle from initiating luteolysis. Nelore (Bos taurus indicus) beef cows (n=40) were submitted to a GnRH/PGF(2alpha)/GnRH protocol. Cows that ovulated from a dominant ovarian follicle (ovulation=Day 0) were allocated to receive: no additional treatment (G(C); n=7); 3000IU of hCG on Day 5 (G(hCG); n=5); 5mg of estradiol-17beta on Day 12 (G(E2); n=6); or 3000IU of hCG on Day 5 and 5mg of estradiol-17beta on Day 12 (G(hCG/E2); n=5). Ultrasonographic imaging of the ovaries, assessment of plasma progesterone concentration, and detection of estrus were done daily from Day 5 to the day of subsequent ovulation. Treatment with hCG induced an accessory CL, increased CL volume, and plasma progesterone concentration throughout the luteal phase (P<0.01). Estradiol-17beta induced atresia and recruitment of a new wave of follicular growth; it eliminated a potentially estrogen-active, growing ovarian follicle within the critical period for maternal recognition of pregnancy, but it also hastened luteolysis (Days 16 or 17 vs. Days 18 or 19 in non-treated cows). In conclusion, the approaches tested enhanced luteal function (hCG) and altered ovarian follicular dynamics (estradiol-17beta), but were unable to extend the life-span of the CL in Nelore cows.     

Induction of precocious puberty in ewe lambs by pulsatile administration of GnRH

Circhoral administration (250 ng/h, i.v.) of GnRH induced a preovulatory-like surge of LH and subsequent luteal function in 4 of 4 ewe lambs 1 month before expected date of puberty. Within 12h of the start of pulsatile delivery of GnRH, mean concentrations of immunoactive and bioactive LH increased significantly (P less than 0.05) and the LH surge occurred by 1.8 +/- 0.6 days of treatment. Mean concentrations of serum progesterone were elevated significantly (P less than 0.001) 3 days after the surge. The biopotency of LH (bioactive LH/immunoactive LH) before the GnRH-induced surge of LH did not differ from LH biopotency in ewe lambs receiving circhoral delivery of saline (0.41 +/- 0.05 and 0.46 +/- 0.04, respectively). Biopotency of LH declined markedly at the GnRH-induced LH surge (0.25 +/- 0.04), but biopotency of serum LH was significantly augmented (P less than 0.05) during the period of luteal activity (0.70 +/- 0.07). Regular oestrous cycles were observed in 3 of 4 ewe lambs after the 10-day GnRH treatment period. These results indicate that pulsatile delivery of GnRH is effective in inducing precocious puberty in ewe lambs. Increase in LH biopotency does not appear to be required in the pubertal transition to reproductive cyclicity in this species. Augmented LH biopotency may be important in support of luteal function after first ovulation.     

Use of a homologous radioimmunoassay (RIA) to evaluate the effect of maternal and foetal parameters on pregnancy-associated glycoprotein (PAG) concentrations in sheep

Early pregnancy detection and prediction of the number of lambs would be profitable for sheep breeders because it enables them to adjust nourishment of pregnant ewes according to the individual needs in order to prevent health problems around parturition. The concentration of ovPAG has previously been reported to be related with maternal parameters (farm, breed and age) as well as foetal parameters (number of lambs, their sex and birth weight), but contradictory results were obtained in different small-scale studies. This large-scale study evaluates the effect of these parameters on the ovPAG concentration, determined by a homologous radioimmunoassay (RIA), and it further investigates the possibility to predict the number of lambs by means of homologous ovPAG determination. Eighty-three and ninety-five ewes of the Suffolk and Texel breed, respectively, housed on four different farms (experiment 1) and 68 ewes of the Suffolk breed, housed on two different farms (experiment 2) were included in this study, and their estrous cycles were synchronised using a progesterone analogue. On the day of synchronisation (D-14) and at 25 (D25), 35 (D35) and 45 (D45) days after insemination, blood samples were taken and a homologous radioimmunoassay (RIA) was used to determine the ovPAG concentrations. At parturition, age of the ewe, number and sex of the lambs (experiment 1) and birth weight (experiment 2) were registered. OvPAG concentrations were not affected by age of the ewe and sex of the lambs. Farm and breed of the ewes, number and birth weight of the lambs had a significant effect on ovPAG concentrations at all time points (P<0.05). The odds of multiple lambs increased significantly with increasing ovPAG concentration, although prediction of litter size based on ovPAG concentration at the individual ewe level was not useful due to small sensitivity and/or specificity whatever the cutoff value used. In conclusion, the ovPAG concentration is affected by farm and breed of the ewes, and number and birth weight of the lambs.     

Serum luteinizing hormone, growth hormone and insulin-like growth factor-I after releasing hormone challenge in prepubertal ewe lambs selected for twinning

Two studies evaluated hormonal markers as indicators of the onset of puberty in Debouillet sheep selected for twinning. In Trial 1, 29 ewe lambs (50 +/- 0.5 kg, 159 to 187 d of age) were given 10 microg GnRH (i.v.) on September 15 and blood was collected at 30 min intervals after the injection for 2 h. Additional samples were taken twice weekly and progesterone (P4) was measured. The day that serum P4 was greater than 1 ng/mL for 2 consecutive sampling days was classified as the day of puberty. Average day of puberty was October 12 (average age at puberty was 199 d) and ewes with values less or greater than the average were classified as early or late, respectively. Average weight at GnRH challenge was 50 kg and ewes weighing less or more were classified as light or heavy, respectively. Early ewes weighed more (P = 0.01) and reached puberty sooner (P = 0.01) than late ewes. Heavy lambs reached puberty earlier, weighed more at GnRH challenge, and had more LH area under the curve (AUC, P < 0.05) than light ewes. In Trial 2, we gave 27 ewe lambs (54 +/- 0.9 kg, 173 to 189 d of age) a single i.v. injection of 10 microg GnRH and 10 microg GHRH on September 17. Average day of puberty was October 13, average weight was 54 kg, and average age at puberty was 208 d. Categories were designated as described for Trial 1. Early lambs reached puberty sooner (P = 0.01) and weighed more (P = 0.01) than late lambs, but the puberty groups had similar LH AUC (P = 0.64) and GH AUC (P = 0.75), whereas IGF-I was greater (P = 0.01) in early puberty ewes than in late puberty ewes. Heavy lambs reached puberty earlier (P = 0.06), weighed more (P = 0.01), and tended (P = 0.11) to have more GH AUC than light ewes. No difference was observed in LH AUC or IGF-I between weight groups (P > 0.15). Results suggest that serum LH after GnRH is not a reliable indicator of the onset of puberty in ewe lambs selected for twinning, but heavier ewes tended to produce more GH after a GHRH challenge and reach puberty earlier than lighter ewe lambs.     

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.     

Growth response, endocrine profiles and reproductive performance of fine-wool ewe lambs treated with ovine prolactin before breeding

Twenty-four 6-mo-old ewe lambs received one of two ovine prolactin (oPRL) treatments 28 d before fall breeding. Beginning on the first day of treatment (Day 0), 12 lambs received a subcutaneous injection (12 ml) of a carrier vehicle (0 mg oPRL) on alternate days for 28 d while 12 lambs received injections containing 5 mg oPRL. On Days 0 and 28, jugular blood was collected from six lambs in each group before treatment and at 30-min intervals for 6 h thereafter. Neither feed intake, efficiency of gain nor animal weights differed (P > 0.20) between groups. One hour after treatment on Day 0, ewe lambs receiving 5 mg oPRL had greater (P < 0.10) serum PRL levels than did controls (121.9 and 61.5 +/- 24.7 ng/ml, respectively). Differences in serum PRL persisted throughout remaining sampling intervals on both Days 0 and 28. Serum samples obtained on alternate days during the 28-d treatment period revealed no differences (P > 0.20) in PRL concentrations between control (48.3 +/- 5.3 ng/ml) and oPRL-treated (55.7 +/- 5.3 ng/ml) ewes. Neither serum insulin nor growth hormone responded (P > 0.05) to exogenous oPRL on either Day 0 or 28. No difference (P > 0.30) in percentage of ewe lambs cycling during treatment or breeding was detected between groups. Subsequent lambing percentages were similar (P > 0.30), with 36.4% of control and 25.0% of oPRL-treated ewes producing offspring. Administering 5 mg oPRL on alternate days for 28 d before breeding did not enhance growth and(or) reproductive performance in virgin ewe lambs.     

Effects of medroxyprogesterone acetate (MAP) on ovarian antral follicle development, gonadotrophin secretion and response to ovulation induction with gonadotrophin-releasing hormone (GnRH) in seasonally anoestrous ewes

When ovulation is induced with gonadotrophin-releasing hormone (GnRH) in anoestrous ewes, a proportion of animals fail to form normal (full-lifespan) corpora lutea (CL). Progesterone treatment before GnRH prevents luteal inadequacy. It remains uncertain whether a similar effect, achieved with medroxyprogesterone acetate (MAP) from intravaginal sponges, is mediated by influences on growing ovarian follicles and/or secretion of gonadotrophic hormones, before and after GnRH treatment. Two experiments were performed, on 13 and 11 anoestrous Western white-faced ewes, respectively. Seven and six ewes, respectively, received MAP-containing sponges (60 mg) for 14 days; the remaining ewes served as untreated controls. To test the effect of timing of GnRH administration after pre-treatment with MAP-releasing sponges, GnRH injections (250 ng every 2h for 24h followed by a bolus injection of 125 microg of GnRH i.v.) were given either immediately (Experiment 1) or 24h after sponge removal in the treated ewes (Experiment 2). Ovarian follicular dynamics (follicles reaching >or=5mm in size) and development of luteal structures were monitored using transrectal ultrasonography. In Experiment 1, the mean ovulation rate (0.7+/-0.3 and 1.0+/-0.4) and proportion of ovulating ewes (57 and 67%, respectively) did not vary (P>0.05) between MAP-treated and control ewes. Normal (full-lifespan) CL were detected in 29% of treated and 67% of control ewes (P>0.05). In Experiment 2, the mean ovulation rate (2.3+/-0.2 and 1.2+/-0.6; P<0.05) and percentage of ewes with normal (full-lifespan) CL (100 and 40%, respectively; P<0.10) were greater in the treated compared to control ewes. In Experiment 1, the mean peak concentration of the GnRH-induced LH surge was lower (P<0.05) in MAP-treated than in control ewes. There were no significant differences between MAP-treated and control ewes in the characteristics of follicular waves, mean daily serum FSH concentrations, and secretory parameters of LH/FSH, based on intensive blood sampling conducted 1 day before sponging and 1 day before sponge removal. It is concluded that treatment with MAP has no effect on the tonic secretion of LH/FSH or follicular wave development in anoestrous ewes. However, the GnRH-stimulated LH discharge was attenuated in the ewes that received MAP-impregnated sponges for 14 days and were treated with GnRH immediately after sponge withdrawal. Ovulatory response and CL formation were increased when GnRH was administered 24 h after sponge removal.     

Effect of oestradiol treatment during GnRH-induced ovulation on subsequent PGF2alpha release and luteal life span in anoestrous ewes

In sheep, induction of ovulation during anoestrus is accompanied by a high incidence of short luteal phases, though pre-treatment with progesterone can overcome this problem. We have investigated the effects of supplementing oestradiol during GnRH-induced ovulation on subsequent PGF2alpha release and luteal life span. Thirty anoestrous crossbred ewes received 250 ng GnRH i.v. at 2 h intervals for 48 h to induce ovulation either alone (group 1; n=10) or in association with either an i.m. injection of 20 mg progesterone 3 days earlier (group 2; n=10) or 3 i.m. injections of 10 microg oestradiol at 8 h intervals on the second day of GnRH treatment (group 3; n=10). Laparoscopy, performed 3 days following GnRH to confirm ovulation and 8 days later, coupled with plasma progesterone analysis were used to determine luteal life span. On day 4 following GnRH, plasma samples were collected at 20 min intervals for 8 h to monitor PGF2alpha release. One ewe from group 1 failed to ovulate and was excluded from further analysis. All groups showed an increase (P<0.01) in plasma oestradiol during GnRH treatment, with group 3 showing a marked (P<0.001) increase over that seen in the other two groups. In group 1 there were 1.4+/-0.2 PGF2alpha episodes/ewe/8 h. In group 2, pre-treatment with progesterone caused the complete inhibition of PGF2alpha episodes (0 episodes/ewe/8 h) while in group 3, treatment with oestradiol resulted in a significant reduction (0.3+/-0.1 episodes/ewe/8 h) compared with group 1 (P<0.01). In group 1, 9/9 ewes exhibited short cycles compared with 2/10 ewes in group 2 (P<0.01). In group 3 the proportion of ewes showing short cycles 7/10 ewes was not significantly different from the other groups. While treatment with oestradiol caused a significant attenuation of PGF2alpha release, this was associated with only a partial reduction in the incidence of short cycles.     

Effects of melengestrol acetate and P.G. 600 on fertility in Rambouillet ewes outside the natural breeding season

The effects of melengestrol acetate (MGA) and P.G. 600 on ewe fertility outside the natural breeding season were evaluated. Rambouillet ewes were assigned to one of four groups: (1) control (C; n=92); (2) PG600 (n=86); (3) MGA (n=99); and (4) MGA+PG600 (n=92). A pellet with or without MGA (0.3mg/ewe/d) was fed at 0.15kg/ewe/d for 7d. On the last day of pellet feeding, ewes were given either saline or 5mL of P.G. 600 i.m. (400IU equine chorionic gonadotropin (eCG) and 200IU human chorionic gonadotropin (hCG)). Ultrasonography was performed between Days 20 and 25 of gestation for ewes that were mated during the first 6 d of the breeding period from the MGA (n=15) and MGA+PG600 (n=8) groups, and the number of luteal structures and embryos were counted. During the first 6d of the breeding period, MGA increased (P<0.05) the percentage of ewes that mated and conceived when compared to C and PG600 (24.2% vs. 3.3% and 10.5%, respectively). Relative to MGA, the mean (+/-S.E.M.) number of luteal structures per ewe was enhanced (P<0.03) in MGA+PG600 (1.53+/-0.13 vs. 2.38+/-0.42, respectively), however as pregnancy progressed, the number of embryos (1.5+/-0.13 vs. 1.8+/-0.16, respectively) and lambs born (1.3+/-0.15 vs. 1.5+/-0.27, respectively) did not differ. Treatment with MGA reduced (P<0.01) the interval from ram introduction to lambing relative to groups that did not receive MGA (168+/-0.8d vs. 171+/-0.6d, respectively). In conclusion, treatment with MGA increased the percentage of ewes conceiving early in the breeding period. Although P.G. 600 increased the number of luteal structures present per ewe, it did not significantly enhance ewe prolificacy.