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.     

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.     

Influence of exogenous melatonin and altered day length on reproductive performance of Polypay ewes

Two experiments were designed to evaluate postlight treatment use of melatonin as a method of overcoming photorefractory response by measuring the lambing percentage of early-postpartum Polypay ewes bred either late in the breeding season or during anestrus. In Trial 1, pregnant ewes (n = 140) were assigned to one of three groups: 1) ambient photoperiod (control), 2) extended light (20 h) from October 21 to December 27, and 3) extended light as in Group 2, followed by supplemental feeding of 10 mg melatonin/head (hd) daily until April 6. Breeding started on February 3 and ended April 9. A greater percentage of ewes given extended light plus melatonin (54%) and extended light alone (45%) lambed than controls (24%) (P = 0.06). In Trial 2, pregnant ewes (n = 158) were assigned to groups as in Trial 1, except extended light was given to Groups 2 and 3 from January 1 to March 11, and Group 3 ewes were supplemented with 10 mg melatonin daily from March 12 to June 10. Breeding started April 18 and ended June 10. Lambing percentage was increased (P < 0.01) by extended light plus melatonin (54%) compared to controls (6%) or ewes given only extended light (10%). The shift from artificially extended light to the shorter ambient light with or without melatonin enhanced the lambing percentage of early postpartum ewes on an accelerated lambing program during breeding late in the season. However, only the more pronounced shift from long days to short days, accomplished with extended light plus melatonin, was effective in inducing out-of-season breeding in Polypay ewes.     

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.     

GnRH or eCG treatment fails to restore reproductive function in GnRH immunized ewes

This study was designed to evaluate the potential of using eCG or GnRH in restoring reproductive functions in GnRH immunized ewes. Thirty-three multiparous Kivircik ewes were randomly assigned into either control group (n=11) or immunization group (n=22). Ewes were immunized against GnRH by injecting with a cocktail of ovalbumin-LHRH-7 (ovalbumin-GnRH-7) and thioredoxin-LHRH-7 (thioredoxin-GnRH-7) fusion proteins generated by recombinant DNA technology in April. 500 IU eCG or 0.008 mg GnRH analogue was used to induce ovulations. Serum GnRH antibodies were present in animals of the immunized group beginning the second week after the first immunization and maintained throughout the study (14 months). Immunization caused anestrus in immunized ewes. eCG or GnRH analogue administration given after 14 days progestagen (20 mg fluorogestone acetate, FGA) treatment during breeding season (mid July) did not induce ovulation in these ewes. Two more attempts with single or multiple eCG injections failed to induce ovulation in this group as well. It appears that the gonadotropin stimulation was not of adequate time since neither eCG nor GnRH administration was able to restore reproductive function in immunized animals. The immunization effect lasted more than a year. These results suggest that GnRH immunization exerts its effect via the hypothalamo-pituitary axis and that more than such stimulation is required to overcome the reproductive suppression.     

Evaluation of GnRH treatment 12 days after AI in the reproductive performance of dairy cows

The objective of this study was to evaluate the effects of GnRH administered at Day 12 post-AI on the reproductive performance of dairy cows. Holstein-Friesian dairy cows (n=103) on a large Hungarian dairy farm were allocated randomly to treated (n=54) or control (n=49) groups. Twelve days after AI, treated cows received a GnRH agonist i.m., while the control group received a placebo (physiological saline). Progesterone radioimmunoassay was used to determine the correct timing of artificial insemination (Day 0) and the incidence of luteal insufficiency on Day 12. Ultrasonography and radioimmunoassay for pregnancy-associated glycoprotein were used to detect pregnancy and late embryonic/fetal mortality between Days 32 and 55 after AI. Three cows from each group were inseminated when progesterone concentrations were >1.0 ng/mL, and six cows (four from the treated and two from the control group) had luteal insufficiency (progesterone<1.0 ng/mL) on Day 12. Late embryonic/fetal mortality occurred in three treated cows and in two control cows. When these cows were removed from the model, calving rates after first service were 59.6% (28/47) and 59.1% (26/44) for treated and control cows, respectively (P>0.05). There was no significant difference between treated and control cows when they were inseminated before or after Day 100 from calving. In summary, administration of a GnRH agonist on Day 12 after AI did not improve reproductive performance in dairy cows. However, our approach may be used for the field evaluation of different treatment protocols.     

Effects of fasting ewes before mating on their reproductive performance

Border Leicester x Merino ewes were fasted for the first six days or the second six days of the oestrous cycle preceding mating in the fall season. When not fasting, ewes were grazed with a control group on irrigated pasture. Compared with control ewes, fasting had no effect on oestrous occurrence or fertilization but did reduce the ovulation rate, especially where fasting occurred early in the oestrous cycle.     

Response of seasonally anoestrous ewes to six-hour periods of GnRH infusion administered on six consecutive days

Twelve seasonally anoestrous Clun Forest ewes were infused i.v. with either 500 or 1000 ng GnRH/h for 6 h on each of six consecutive days in early or mid-anoestrus. Plasma LH concentrations were elevated during each GnRH infusion but returned to pretreatment levels when infusion ceased. The response to the first infusion was significantly greater (p<0.001) than that to subsequent infusions. In addition, both a GnRH dose and a seasonal influence were evident in the LH response, but these failed to reach statistical significance. Although 7 12 ewes ovulated, only two produced functionally normal corpora lutea.     

Effects of ageing and exogenous melatonin on pituitary responsiveness to GnRH in ewes during anestrus and the reproductive season

The study examined the effect of melatonin implants on in vivo pituitary responsiveness to GnRH in control, fully productive (5.7+/-0.4 years old, n=17) and aged (10.7+/-0.3 years old, n=14) ovariectomized, estradiol-treated Rasa Aragonesa ewes. On 27 February, eight ewes in each age group received a single implant containing 18 mg melatonin. On 10 April, blood samples to be assayed for LH were collected at 10-min intervals over 4h (starting at 09:00 and 22:00 h). After samples 6 and 18 were collected, ewes received a single i.v. injection of GnRH (20 ng/kg liveweight). The pituitary response to GnRH was assessed using the difference between plasma LH concentrations before and after (highest value) each injection (DLH1, DLH2)), and the area under the LH response curve for 1h after each GnRH injection (AUC1, AUC2). On 23 September, the previously implanted ewes received a new melatonin implant and, on 17 November, all of the ewes were subjected to the same diurnal and nocturnal sampling protocols, again. Generally, non-implanted aged ewes exhibited a lower pituitary response to GnRH than did non-implanted control ewes, particularly in November and after the first injection (P<0.05 for DLH1 and AUC1 in both the diurnal and nocturnal tests). The response was significantly affected by the interaction of age and melatonin treatment, particularly in the diurnal tests (P<0.1 for DLH1 and AUC1, and P<0.05 for AUC2 in April; P<0.05 for DLH1, AUC1 and AUC2 in November), which indicated that exogenous melatonin increased LH levels after GnRH injections in aged ewes compared to non-implanted ewes, this effect being the opposite in control females. Thus, melatonin can restore in ewes the functionality of the neuroendocrine system, after it has been reduced by senescence.     

Gonadotropin secretion in ovariectomized ewes: effect of passive immunization against gonadotropin-releasing hormone (GnRH) and infusion of a GnRH agonist and estradiol

Gonadotropin secretion was examined in ovariectomized sheep after passive immunization against gonadotropin-releasing hormone (GnRH). Infusion of ovine anti-GnRH serum, but not control antiserum, rapidly depressed serum concentrations of luteinizing hormone (LH). The anti-GnRH-induced reduction in serum LH was reversed by circhoral (hourly) administration of a GnRH agonist that did not cross-react with the anti-GnRH serum. In contrast, passive immunization against GnRH led to only a modest reduction in serum concentrations of follicle-stimulating hormone (FSH). Pulsatile delivery of the GnRH agonist did not influence serum concentrations of FSH. Continuous infusion of estradiol inhibited and then stimulated gonadotropin secretion in animals passively immunized against GnRH, with gonadotrope function driven by GnRH agonist. However, the magnitude of the positive feedback response was only 10% of the response noted in controls. These data indicate that the estradiol-induced surge of LH secretion in ovariectomized sheep is the product of estrogenic action at both hypothalamic and pituitary loci. Replacement of the endogenous GnRH pulse generator with an exogenous generator of GnRH-like pulses that were invariant in frequency and amplitude could not fully reestablish the preovulatory-like surge of LH induced by estradiol.     

Ovarian responses of seasonally anestrous ewes administered progesterone,PMS, HCG and(or) GnRH

One hundred and sixty ewes were assigned to sixteen groups in a 2 × 2 × 4 factoral design and were treated during the anestrous season. The main effects were progesterone pretreatment (non-implanted and implanted for 14 days), PMS pretreatment (no pretreatment and pretreatment with 500 IU at the time of progesterone implant removal) and treatments (none, GnRH in saline, GnRH in gelatin capsules and HCG). GnRH in saline (250 μg) and HCG (500 IU) were administered intramuscularly and GnRH in gelatin capsules (250 μg) was administered subcutaneously 24 hours after the time of progesterone implant removal.Ewes were classified into one of four progesterone response categories: cyclic, transient, prolonged and no response. An injection of GnRH in saline induced a prolonged progesterone response in only one ewe (13%) which was similar to the response in the untreated ewes (0%). More ewes administered GnRH in gelatin capsules (56%) and more ewes administered HCG (89%) had a prolonged progesterone response than GnRH (in saline) treated or untreated ewes. A higher percentage of ewes that were pretreated with PMS and treated with GnRH in saline (78%) had a prolonged progesterone response than ewes treated with either PMS (22%) alone or with GnRH (in saline; 13%) alone.     

Effect of insulin on feed intake and reproductive performance of well-nourished nulliparous ewes

Eighty-four nulliparous ewes were used to examine the effect of short-term insulin treatment on feed intake and reproductive performance. Following estrus synchronization, ewes were observed for estrus (= Day 0) and were penned individually beginning on Day 7. Ewes were fed twice daily and feed intake was recorded. On Days 9 through 13, ewes were treated s.c. with 1 IU/kg BW insulin (n = 44) or an equivalent volume of saline (n = 40). On Day 14, ewes were placed with fertile rams and number of ewes in estrus (bred) was recorded. Thirty days post-breeding, ewes were checked for pregnancy via ultrasonography. Feed intake and percentage of ewes in estrus did not differ between saline- and insulin-treated ewes. Similarly, neither pregnancy rate (69 +/- 8.7% vs. 80 +/- 8.1%, respectively) nor lambing rate (61 +/- 8.9% vs. 78 +/- 8.4%, respectively) differed between treatments. The number of lambs born per ewe was, however influenced by a breed-group effect (P < 0.0002). Romanov ewes had more (P < 0.001) lambs than the other breed groups in the study. Therefore, treating well-nourished, nulliparous ewe lambs with insulin did not increase reproductive efficiency, possibly because the ewes were already at a maximal nutritional and/or reproductive state.     

Effect of zinc supplementation on the reproductive performance of grazing Merino ewes

Two experiments were carried out in Western Australia to investigate the effect of zinc supplementation, on the reproductive performance of grazing Merino ewes. We found that supplemental zinc, when provided prior to mating and throughout pregnancy, increased the number of lambs produced by 14% (P<0.05) in both experiments. An intermediate zinc treatment, when supplementation was begun later in pregnancy gave a 9% (P<0.01) increase in the number of lambs in one experiment and no increase in the second. Lamb birth weights were increased by zinc supplementation in Experiment 1 and in Experiment 2, 12–20-week-old lambs from zinc supplemented ewes were 2.1 kg heavier than those from nonsupplemented controls. Plasma zince levels decreased significantly during pregnancy and lactation, but were increased at some sampling dates by 20–25% by zinc supplementation.     

Ultrasonography of the developing reproductive tract in ram lambs: effects of a GnRH agonist

In spring-born ram lambs, the testes (from 2 wk), prostate and vesicular glands (from 4 wk) were examined by ultrasonography every 2 wk up to 26 wk of age. Image analysis was done (numerical pixel values). Ram lambs were treated with a long acting formulation of a GnRH superagonist (Leuprolide acetate; 1.5 mg/kg) at 3 and 7 wk of age. In blood samples taken every 15 min for 8 h, mean serum LH, LH pulse amplitude, and basal and mean serum FSH concentrations were lower at 5 wk of age, and LH pulse frequency was lower at 15 wk of age in animals given Leuprolide acetate compared with those of the controls. There were no differences (P>0.05) in testis, prostate or vesicular gland development between treated and control animals. Testicular diameter of the left and right testes in transverse and longitudinal planes increased slowly to 8 wk of age, more rapidly to 18 wk of age, then more slowly to 26 wk of age (P<0.05). Numerical pixel values of testicular images decreased from 2 to 8 wk of age, increased to 22 wk of age and then plateaued. Width of the prostate increased from 4 to 26 wk of age, but length and width of the vesicular glands increased slowly to 8 wk of age, more rapidly to 18 wk of age and then plateaued (P<0.05). Numerical pixel values for the prostate declined from 4 to 8 wk and for the vesicular glands, declined from 4 to 10 wk of age; numerical pixel values increased to 12 wk and then decreased to a nadir at 18 wk, followed by a steady increase to 26 wk of age (P<0.05). We concluded that developmental patterns of numerical pixel values of the testes, prostate and vesicular glands in ram lambs reflect stages of development, but treatment with a GnRH superagonist at 3 and 7 weeks of age did not affect growth of testes, vesicular or prostate glands.     

Effects of retention of fetal membranes on subsequent reproductive performance of dairy ewes

Seventy-one pairs of ewes with or without retention of fetal membranes in the previous lambing season, were included in a cohort study of 25 flocks in southern Greece; in 27 pairs regulation of the breeding cycle was applied, whilst in the other 44 pairs natural mating took place. The following measures of reproductive performance were calculated: cycling rate, mating rate, return-to-estrus rate, abortion rate, lambing rate, total lambs born per ewe, liveborn lambs per ewe, stillbirth rate, lamb-bodyweight per ewe; furthermore, the incidence risk of retention of fetal membranes during the lambing examined in this study was also calculated. No statistically significant differences were observed in the reproductive performance among ewes that had or had not retained their fetal membranes during the previous lambing, whether the breeding cycle was regulated or not. The incidence risk of retention of fetal membranes among ewes that had not retained its placenta in the previous lambing was 1.6%, whilst that among ewes that had retained its placenta in the previous lactation was 0%. It is concluded that retention of fetal membranes did not appear to adversely affect subsequent reproductive performance of ewes.     

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

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

The effect of treatment with a GnRH analogue on postpartum reproductive performance in Friesian cows

Three trials were conducted to examine the effect of GnRH injection on the reproductive performance of Friesian cows. In the first trial 100 μg gonadotrophin releasing hormone synthetic analogue (GnRH) was given at the time of A.I. to 32 cows while a control group received a saline placebo injection. Conception rate to first insemination was significantly higher in treated than control cows (81.3 v. 54.8%, P < 0.05).In the second trial, two groups of 19 cows each received either 100 μg GnRH or saline at 15 days postpartum. A total of 60 cows was used in the third trial in which GnRH was given at either 50 or 100 μg dose level on either days 7 or 15 postpartum while control cows were untreated. In both trials GnRH treatment reduced the intervals from calving to complete uterine involution, first ovulation and first detected oestrus. The effect was most pronounced when 100 μg GnRH was given on day 7 postpartum. Number of days open and number of services per conception were appreciably reduced when 100 μg GnRH was given at either 7 or 15 days postpartum with maximal effect when given 15 days after calving.     

Pattern of gonadotropin-releasing hormone (GnRH)-like stimuli sufficient to induce follicular growth and ovulation in ewes passively immunized against GnRH.

The pattern of GnRH-like stimuli capable of inducing follicular growth, ovulation, and luteal function was evaluated in ewes passively immunized against GnRH. The estrous cycles of 30 regularly cyclic sheep were synchronized using vaginal pessaries impregnated with a synthetic progestogen. Animals were passively immunized against GnRH (groups 2-5, n = 6) or the carrier protein, keyhole limpet hemocyanin (KLH; group 1, n = 6), at the time of pessary removal (PR). Circhoral delivery of saline (groups 1, 2, and 5) or low amplitude GnRH agonist (des-Gly10 GnRH ethylamide [100 ng/hourly pulse]; groups 3 and 4) was initiated at PR and continued for 3 (groups 4 and 5) or 12 days (groups 1-3). In groups 4 and 5, the amplitude of the GnRH-like stimulus was increased to 800 ng/hourly pulse (stimulus-shift) during the 24-h period beginning 72 h after PR. The amplitude of the hourly stimulus was adjusted to 100 ng/pulse 96 h after PR and continued at that level to Day 12. The endocrine changes associated with follicle growth and maturation (serum concentrations of estradiol [E2] above 10 pg/ml), ovulation (surge-like secretion of LH and FSH), and normal luteal function (serum concentrations of progesterone [P] above 2 ng/ml) were evident in ewes passively immunized against KLH (group 1). In this group, the preovulatory surge of gonadotropins was noted 48.7 +/- 1.2 h after PR. These endocrine events were blocked by passive immunization against GnRH (group 2).(ABSTRACT TRUNCATED AT 250 WORDS)