Jugular levels of 13,14-dihydro-15-keto-prostaglandin F and progesterone around luteolysis and early pregnancy in the ewe

Six non-pregnant ewes at day 12 of the estrous cycle each had a day-12 embryo transferred into the uterine horn ipsilateral to the corpus luteum, and 4 non-pregnant ewes at day 13 each had a day-13 embryo similarly transferred. Four control ewes, 2 at day 12 and 2 at day 13 received sheep serum into the uterine horn ipsilateral to the corpus luteum. Jugular blood samples were taken at 2-hourly intervals for 3 days post-surgery, then twice-daily for a further 4 days, and the plasma radioimmunoassayed for progesterone and 13,14-dihydro-15-keto-prostaglandin F. All control ewes exhibited estrus within the expected time range and pulsatile peaks of 13,14-dihydro-15-keto-prostaglandin F occurred coincident with declining progesterone levels. With one exception, the recipient ewes had prolonged cycles and those ewes found pregnant at necropsy, 30 days after transfer, showed no progesterone decline and no pulsatile peaks of prostaglandin during days 12 to 16 after estrus. These observations suggest that the presence of the embryo at a critical stage after mating suppresses the release of uterine prostaglandin F_2α.     

Effect of prostaglandins on uterine contractions in the estrous ewe.

Administration of exogenous prostaglandins at the time of mating may improve fertility via their effects on uterine contractility. The present study was undertaken to compare the effects of three prostaglandins that affect either the male or female reproductive uterine contractility. Contractions in the uterine body of anesthetized ewes during estrus were studied before, during and after a 5 min interval of systemic infusion of prostaglandin F_2α-THAM salt (PGF_2α; 5 mg), prostaglandin E₁ (PGE₁; 5 mg), prostaglandin E₂ (PGE₂; 5 mg) or vehicle. Pressure changes were detected by the use of an open-ended intrauterine catheter and a transducer. Each of the three prostaglandins initially caused a single prolonged contraction that lasted about 10 minutes and had a maximum pressure of 50 mm Hg. Prior to the prolonged contraction, PGE₁ and E₂ caused a relaxation for about 1 minute. In addition, PGE₁ and E₂ caused more secondary contractions (15–20) during the prolonged contraction than did PGF_2α (7–9). The effects of prostaglandin (PG) treatment lasted for 20–30 minutes. The authors conclude that with the dose used the three prostaglandins studied do not have greatly different effects on uterine contractility in estrous ewes.     

Sheep breeding schemes utilising artificial insemination; large-scale simulation with a complex breeding goal

Alternative Norwegian sheep breeding schemes were evaluated by stochastic simulation of a breeding population with about 120 000 ewes, considering the gain for an aggregate genotype including nine traits and also the rate of inbreeding. The schemes were: a scheme where both young unproven rams (test rams) and proven rams (elite rams) are used in artificial insemination (AI scheme), a scheme with test rams in natural mating in ram circles and elite rams (from one and a half years of age) in AI across all flocks in the country (NMAI2 scheme), a scheme where, in addition to testing rams, the youngest elite rams (one and a half years of age) are also used in natural mating in ram circles, while older elite rams are used in AI (NMAI1 scheme), and a scheme, acting as a control, where both test and elite rams are used in natural mating (NM scheme). Within the NMAI- and AI-schemes, experimentation was performed for percent ewes inseminated to elite rams v. test rams (EM%), numbers of ewes inseminated per elite ram (EAIn), and numbers of ewes mated per test ram by natural service (TNMn) or by AI (TAIn), respectively. With a restriction on the rate of inbreeding (⩽0.8% per generation), the AI scheme gave similar gain to the NMAI2 scheme (and about 40% more than did the NM scheme). Less gain was generated by the NMAI1 scheme, but it was still considerably more than for the NM scheme (about 25%). In the AI scheme, relatively few ewes (200/300) should be inseminated to each test/elite ram, and a low EM% should be chosen (10%). In the NMAI schemes, TNMn should be relatively high (40 to 50), combined with average and somewhat larger than average EAIn (NMAI2: 700 ewes, NMAI1: 900 ewes), and EM% medium (30%).     

Seasonal variations in prolactin, growth hormone and thyroid hormones and the prolactin surge at ovulation do not affect litter size of ewes during pregnancy in the oestrous or the anoestrous season

Injection of bromocriptine from 5 days before until 5 days after mating clearly suppressed the periovulatory prolactin surge in ewes in the anoestrous and oestrous season but did not change the litter size significantly. Progesterone, GH, TSH or thyroid hormone concentrations were not influenced by the bromocriptine treatment. The progesterone concentrations were lower during the first weeks after mating in the anoestrous season compared to the oestrous season, while there was no difference between pregnant and non-pregnant ewes. During later gestation this seasonal difference was only observed in the non-pregnant ewes. At the same time there was a clear difference between pregnancy and non-pregnancy in both seasons. The prolactin, GH and thyroid hormone values also varied significantly during gestation. Since these patterns are identical in pregnant and non-pregnant ewes, the fluctuations are due to environmental factors and not to pregnancy or altered progesterone concentrations. In the anoestrous season prolactin, GH, T4 and T3 levels were higher than in the breeding season, while rT3 showed the opposite pattern. The TSH concentration did not differ between the two seasons. These results suggest that seasonal variations in prolactin, GH and thyroid hormones or the periovulatory prolactin surge do not affect litter size of ewes during pregnancy in the oestrous or the anoestrous season.     

Successful artificial insemination for indoor breeding in the Japanese monkey (Macaca fuscata) and the cynomolgus monkey (Macaca fascicularis)

Methods of artificial insemination (AI) for indoor breeding in the Japanese monkey and the Cynomolgus monkey were investigated. For the Japanese monkey AI was carried out in six females during the winter mating season and in six females during the summer non-mating season. During the mating season, semen was inseminated near ovulation time in natural menstrual cycles. In the mating season study, three females inseminated at the uterine cavity became pregnant. Three inseminated at the cervical canal failed to become pregnant. For the non-mating season study, ovulation was induced artificially by PMSG and hCG and AI was carried out near the induced ovulation time. In the non-mating season, no animals became pregnant. Of four Cynomolgus monkeys used, pregnancy occurred in two animals inseminated near ovulation time in natural menstrual cycles. AI occurred at the uterine cavity in one and cervical canal in the other. In both species ovulation was verified by laparoscopy. Semen was collected by penile electro-stimulation then diluted to 2.5 to 5.0×10⁷/ml with Whitten's medium. Diluted semen of 0.2l was inseminated at the uterine cavity or cervical canal. Our results indicate the usefulness of vaginal AI as a method of artificial indoor breeding.     

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.     

Investigations on the detrimental effect of prostaglandin F2α-regulated estrus on the number and condition of sperm in the reproductive tract of the ewe

Ewes in the luteal phase of the estrous cycle were treated with prostaglandin F₂α (PGF), mated to rams at the ensuing estrus 2 days later, and necropsied at 2 or 23 hr after mating. At 2 hr after mating, ewes in PGF-regulated estrus had significantly fewer sperm in the middle and anterior one-thirds of the cervix and in the uterus than did ewes mated during natural estrus. At 23 hr, soon after ovulation, significantly fewer ewes in PGF-regulated estrus had sperm in the oviducts than did ewes in natural estrus.In Experiment 2, ewes in PGF-regulated or natural estrus were laparotomized, inseminated by deposition of semen in the uterine lumen, and necropsied 2 or 23 hr later. Intrauterine insemination prevented most of the reduction in sperm numbers in the reproductive tract at PGF-regulated estrus.In Experiment 3, ewes in PGF-regulated or natural estrus were either mated to rams or inseminated in the uterine lumen and necropsied 2 hr later. Sperm were recovered from three segments of the cervix and were counted and evaluated for motility, response to live-dead staining, and acrosomal morphology. Intrauterine insemination again reduced the detrimental effect of PGF-regulated estrus on sperm numbers. However, the percentages of sperm recovered from the cervix that were motile, live, and had normal acrosomes were much lower in ewes in PGF-regulated estrus than in ewes in natural estrus. Compared with natural mating, intrauterine insemination reduced but did not eliminate the detrimental effects of PGF-regulated estrus on the viability and morphology of sperm. Regulating estrus with PGF resulted in damage to sperm in the cervix regardless of whether sperm reached the cervix from the vagina or from the uterus.     

Oestrogenic effects of ICI 182,780, a putative anti-oestrogen,on the secretion of oxytocin and prostaglandin F2α during oestrous cycle in the intact ewe

The effect of ICI 182,780, oestrogen antagonist, on the concentrations of oxytocin and uterine PGF_2α was investigated in intact Border Leicester Merino cross ewes during the late oestrous cycle. Twelve cyclic ewes (n=6 per group) were randomly assigned to receive, at 6 h intervals, intra-muscular injection of either peanut oil or ICI 182,780 (1.5 mg kg⁻¹ day⁻¹) in oil for 2 days, starting at 1900 h on day 13 until 1300 h on day 15 post-oestrus. Hourly blood samples were collected via a jugular catheter from 0800 h on day 14 for 37 h and then daily over days 16, 17 and 18 post-oestrus. Peripheral plasma concentrations of oxytocin, the metabolite of prostaglandin F_2α, 15-keto-13,14-dihydro-prostaglandin F_2α, (PGFM) and progesterone were measured by radioimmunoassay. All ewes treated with ICI 182,780 exhibited functional luteal regression as indicated by a marked reduction in plasma progesterone concentrations to less than 1000 pg/ml over the period of 18–36 h during sampling period on days 14 and 15 of the oestrous cycle. In five of six vehicle-treated ewes, progesterone concentrations declined between day 16 and day 18 post-oestrus. In the remaining control ewe, progesterone concentrations reach less than 1000 pg/ml within 36 h of the commencement of the sampling period. During the frequent sampling period, the number of oxytocin pulses in the ICI 182,780 treated ewes was significantly higher compared to control ewes (2.7±0.3 vs. 0.8±0.3). The mean amplitude of oxytocin pulses observed was also greater (70.4±19.5 pg/ml) in ewes treated with ICI 182,780, but was not significantly different from control ewes (33.5±12.9 pg/ml). Oxytocin pulses may however have occurred following the initial two ICI 182,780 injections but before commencing blood sampling. The oxytocin pulses were detected at a mean of 3.2±0.2 h following each injection with ICI 182,780 during blood sampling. In the ICI 182,780-treated ewes, the pulsatile pattern of plasma PGFM in jugular blood samples over the 37 h sampling period on days 14 and 15 post-oestrus had a higher amplitude (512.9±158.9 vs. 121.7±78.7 pg/ml) and pulse area (618.1±183.3 vs. 151.5±102.9 (pg/ml)τ) compared to the vehicle-treated ewes (P<0.05) respectively. The average number of PGFM pulses observed per ewe was 3.0±0.7 in the ICI 182,780-treated group and was significantly (P<0.02) higher than the number of pulses (0.5±0.3) observed in ewes treated with vehicle alone. The PGFM pulses were detected at 4.2±0.6 h following each injection with ICI 182,780 during blood sampling. The percentage of PGFM pulses that occurred coincidently with a significant elevation of oxytocin concentrations was 44.4% in ICI 182,780-treated compared to 66.6% in control ewes. We conclude that administration of oestrogen antagonist ICI 182,780 accelerated development of the luteolytic mechanism by enhancing pulsatile secretion of oxytocin and PGFM which suggests that ICI 182,780 acts as an agonist for oxytocin and prostaglandin F_2α release in intact ewes when administered at 1.5 mg/kg/day over Day 13 to 15 post-oestrus.     

Effect of injected bovine interferon-alpha I1 on estrous cycle length and pregnancy success in sheep

In Exp. 1 twice daily i.m. injections of 2 mg recombinant bovine IFN-alpha I1 (rboIFN-alpha I1) (N = 24) or placebo (N = 25) were administered to ewes from Day 12 to Day 16 during a normal oestrous cycle. Treatment did not increase (P greater than 0.10) oestrous cycle length (20.7 +/- 1.2 versus 18.5 +/- 1.4 days). In Exp. 2, ewes were injected twice daily with 2 mg IFN (N = 34) or placebo (N = 36) from Days 11 to 18 after natural mating. The rboIFN-alpha I1 significantly (P = 0.05) improved pregnancy rate (79% versus 58%) as determined by a failure of ewes to return to oestrus within 50 days. The number of ewes that lambed was greatest in the rboIFN-alpha I1-treatment group (71% versus 50%; P = 0.07), and no teratogenic effects were observed in the young born to IFN-treated ewes. The study was repeated a second year with a more fecund group of ewes (Exp. 3). More (P = 0.08) ewes injected with rboIFN-alpha I1 (58/65) than placebo-treated ewes (48/61) were judged pregnant by ultrasound. Again more ewes lambed (55 versus 45) and more lambs were born (98 versus 80) from the rboIFN-alpha I1-treated group. Combining the data from both studies revealed a significant (P = 0.01) effect of treatment. The amount of antiviral activity in jugular vein blood of ewes injected with rboIFN-alpha I1 (2 mg) was determined over time in Exp. 4. Activity rose to a maximum (approximately 450 IRU/ml) within 1-2 h and declined by over 75% in 24 h. Single injections of 1, 2 and 5 mg in buffer or 2 mg emulsified in sesame oil all gave similar profiles of antiviral activity in jugular blood over a 48-h period. In Exp. 5, antiviral activity was measured in uterine vein, ovarian artery and jugular vein serum of untreated pregnant (N = 7) and non-pregnant (N = 11) ewes at Day 15 after mating. Activity was detected in the uterine vein (58 +/- 19 IRU/ml) of all pregnant ewes. The observations in Exps 1-5 are consistent with a role for conceptus-derived IFN-alpha in maternal recognition of pregnancy and suggest that supplemental IFN-alpha might be useful in improving pregnancy success in sheep.     

Hormone studies on ewes grazing an oestrogenic (Yarloop clover) pasture during the reproductive cycle.

The endocrine function of Merino and Corriedale ewes grazing an oestrogenic (Yarloop clover) pasture has been studied during the oestrous cycle, pregnancy and parturition, and the results compared with those from a study of similar ewes grazing a neighbouring grass pasture. Plasma progesterone, oestrogen and corticoids were measured using competitive protein binding assay procedures. During the oestrous cycle clearly anomalous patterns in hormone content were evident in ewes grazing Yarloop, and this related to their significantly poorer (P less than 0.001) fertility. The first mating, when ewes were 1-1/2 years of age, was particularly affected. Successful conception took place in only 27% of ewes mated on Yarloop, compared with 95% on grass. Evidence of disturbance in the normal patterns of both plasma oestrogen and progesterone was found in infertile ewes, including a shortened period of luteal function. Disturbance of endocrine function caused by Yarloop clover ingestion was also found in pregnant ewes, with the mean plasma progesterone concentrations during the latter half of pregnancy reduced (P less than 0.05) and the plasma oestrogen and corticoid levels tending to be higher in these animals. In detailed hormone studies in the periparturant period, both groups showed a similar fall in plasma progesterone and rise in plasma oestrogen prior to parturition. Where excessive time was taken for parturition (more than 30 min) this was reflected in higher plasma corticoid levels (P less than 0.05) within 8 h of birth.     

Melanin concentrating hormone (MCH) in the cerebrospinal fluid of ewes during spontaneous oestrous cycles and ram effect induced follicular phases

The melanin-concentrating hormone (MCH) is a neuropeptide synthesized by neurons of the lateral hypothalamus and incerto-hypothalamic area that project throughout the central nervous system. The aims of the present report were: (1) to determine if MCH levels in cerebrospinal fluid (CSF) of ewes vary between the mid-luteal and the oestrous phase of spontaneous oestrous cycles; and (2) to study if MCH levels in CSF of ewes vary acutely during the follicular phase induced with the ram effect in anoestrous ewes. In the first experiment, CSF was collected from 8 adult ewes during spontaneous oestrous and during the mid-luteal phase (8-10 days after natural oestrus). In the second experiment, performed during the mid non-breeding season, a follicular phase was induced with the ram effect. After isolating a group of 16 ewes from rams, CSF was obtained from 5 of such ewes (control group). Three rams were joined with the ewes, and samples were obtained 12h (n=6) and 24h (n=5) later. In Experiment 1, there were no differences in MCH concentrations in CSF measured during the mid-luteal phase and spontaneous oestrus (0.14 ± 0.04 vs. 0.16 ± 0.05 ng/mL respectively). In Experiment 2, MCH concentrations tended to increase 12h after rams introduction (0.15 ± 0.08 vs. 0.35 ± 0.21 ng/mL, P=0.08), and increased significantly 24h after rams introduction (0.37 ± 0.15 ng/mL, P=0.02). We concluded that MCH concentration measured in the CSF from ewes increased markedly during the response to the ram effect but not during the natural oestrous cycle of ewes.     

Reproductive performance of Coopworth ewes following oral doses of zearalenone before and after mating

The reproductive performance of Coopworth ewes after administration of zearalenone was determined in two trials. In Trial I, zearalenone was administered to groups of 33 ewes at rates of 0, 1.5, 3.0, 6.0, 12.0 and 24.0 mg/ewe/day for 10 days, starting on Day 7 of the oestrous cycle before mating. There was a linear decline (P less than 0.001) in ovulation rate with dose of zearalenone; also cycle length decreased and duration of oestrous increased with increasing dose levels. Reductions in the incidence of ovulation and in fertilization were seen only at doses of 12 and 24 mg. In Trial 2, groups of 50 ewes were given the same range of doses of zearalenone for 10 days, starting 5 days after mating to entire rams. There was no effect of zearalenone treatment after mating on pregnancy rate or embryonic loss. These results indicate that the effects of zearalenone, administered orally, on ewe reproduction, at the dose levels examined, were restricted to ewes exposed before mating. Intakes of zearalenone of 3 mg/ewe/day or more during this period would be reflected as depressed ovulation rates and lower lambing percentages.     

Cellular proliferation in the lymphoid tissues of an inbred (RT1a) rat strain during the oestrous cycle

Changes in body weight, uterine weight and tissue weight, cell content and cellular proliferation of the thymus and uterine regional and popliteal lymph nodes were examined at daily intervals during the oestrous cycle of the DA (RT1a) inbred rat strain. Thirty-nine sexually mature virgin animals, aged between 13 and 15 weeks were used in this investigation. During dioestrus body weight and uterine weight fell significantly, while intrathymic and intranodal cellular proliferation increased significantly. These findings are discussed in relation to ovarian hormone secretion and it is suggested that increased thymocyte and lymphocyte proliferation occurs in response to rising oestrogen levels. This proliferative response prepares the female for the immunological challenge with allogeneic spermatozoa should mating occur during the subsequent oestrous phase.     

A comparison between intravaginal progestagen and melatonin implant treatments on the reproductive efficiency of ewes

A total of 548 Manchega ewes were used to compare progestagen impregnated intravaginal sponges (12 days) plus eCG with melatonin implant (100 days) treatments, as possible stimulants of reproductive performance during the seasonal anoestrus and the lactation period. The use of both methods of oestrous manipulation induced a significant improvement in conception rate (78% for progestagen and eCG; 78% for melatonin versus 65% for the control) and fecundity (1.50 for progestagen and eCG; 1.55 for melatonin versus 1.26 for the control), when compared to the control group. Likewise, the lambing rate was also increased (1.17 for progestagen and eCG; 1.21 for melatonin versus 0.82 for control). A total of 35 and 39 extra lambs/100 ewes mated were produced, with an increase in productivity (percentage of extra lambs) of 43% and 48%, for ewes treated with progestagen sponges and melatonin, respectively, compared to the control group. The interval between ram introduction and parturition was significantly decreased, with matings in February–April (early seasonal anoestrus), compared to matings in March–May (seasonal anoestrus) in the group treated with progestagen sponges. An increase in the interval between parturition and the following mating was also recorded. As the body condition improved, the number of lambing ewes and fecundity were significantly improved in all groups. All the fertility results obtained with the progestagen sponges and melatonin treatments were similar. It could be concluded that progestagen sponges plus eCG and melatonin implants improved the reproductive performance of ewes, in seasonal anoestrus and during the lactation period, to a similar extent.     

Plasma levels of cortisol and oxytocin,and uterine activity after cervical artificial insemination in the ewe

The objective was to compare in the ewe the effects of easy and difficult procedures for artificial insemination (AI) (as related to rapid or poor accessibility of the cervix, respectively) on plasma cortisol (CORT) and oxytocin (OT), and uterine motility. All AI were simulated using a catheter empty of semen to study genital and environmental stimuli only. In experiment 1, 40 ewes were sampled after Al, and whether it was an easy or difficult procedure was reported for each animal. While CORT concentrations rose to a similar amount in all ewes, whatever the Al procedure, a significant OT response occurred after a difficult procedure only (n = 18) (17.4 +/- 1.7 versus 12.7 +/- 0.7 pg x mL(-1) before Al, p < 0.05). In experiment 2, uterine activity was monitored in 4 ewes using an implantable telemetric transmitter equipped with an intrauterine pressure catheter. An increased uterine activity occurred during 2 +/- 1 min after an easy Al (n = 5), whereas the evoked activity lasted for 15 +/- 4 min after a difficult Al (p < 0.001, n = 7). A similar long-lasting response occurred after OT administration (100 mIU, i.v.). We concluded that the increase in uterine motility after a difficult Al resulted from a reflex release of OT, and not to a "stress" effect.     

Lambing rates and litter size following carazolol administration prior to insemination in Kivircik ewes

The effect of carazolol on the ease of penetrating the cervix during artificial insemination, lambing rate and litter size was studied using 1.5–4.0-year old Kivircik ewes in an incomplete 3 × 2 × 2 experimental design. All of the ewes in this study were synchronized for oestrus by insertion of a progesterone impregnated vaginal sponge for 12 days and administration of 400 IU PMSG at sponge withdrawal. Three methods of service were compared: natural service, artificial insemination (AI) with fresh semen, or AI with frozen semen. Two times of insemination (fixed time AI versus AI at observed oestrus) were compared on the fresh and frozen AI treatments. The absence (control) or use of carazolol (carazolol; 0.5 mg/ewe i.m. 30 min before mating) was the third factor in the design and penetration of the cervix by the insemination pipette was assessed as shallow (<10 mm), middle (10–20 mm) or deep (>20 mm). Natural service ewes were only mated at observed oestrus. Consequently, the factorial design was incomplete and there were a total of 10 treatments each represented by 30 ewes. Natural service resulted in a significantly (P < 0.05) higher lambing rate and litter size (86%; 2.0 ± 0.05 lambs/ewe) than AI using fresh (65%; 1.6 ± 0.1 lambs/ewe) or frozen (40%; 1.4 ± 0.14 lambs/ewe) semen. For AI animals the lambing rate and litter size were not significantly different when service was at a fixed time (50%; 1.5 ± 0.12 lambs/ewe) or at observed oestrus (56%; 1.5 ± 0.12 lambs/ewe). Carazolol did not permit complete cervical penetration in any ewe. Deep penetration of the cervix at AI was achieved in 33% of untreated (control) and 48% of carazolol treated ewes (P < 0.05). However, the proportion of ewes in which penetration of the cervix and semen deposition was greater than shallow was similar for control (82%) and carazolol (85%), and lambing rate and litter size were similar for both treatments. Over the three service methods, the lambing rate was 56% for control and 63% for carazolol (NS) and litter size was similar for both treatments. It was concluded that the carazolol treatment used prior to natural mating or AI in this experiment did not improve lambing rate or litter size in Kivircik ewes.     

Pattern of electrical activity of the ovine uterus and cervix from mating to parturition

The electrical activity of the uterus, greater curvature and lesser curvature, and of the cervix were recorded throughout several oestrous cycles, matings and the entire course of pregnancy in a set of ewes. In cyclic ewes, all electrode sites were active during the periovulatory period, but during the luteal phase only the cervix and lesser curvature continued to present regular activity, i.e. episodes of 6-8 min duration. Matings failed to disrupt the oestrous activity pattern in 5 out of 6 ewes. During the subsequent pregnancy in 4 ewes, activity, when present, consisted of the cyclic occurrence of regular activity episodes in the gravid horn(s) (greater curvature) and in the cervix (and lesser curvature in 1 ewe). Three phases were recognized on the basis of the activity of the greater curvature of the gravid horn(s). The first phase corresponded to quiescence of the horn from Day 5 (Day 0 = mating) to about Day 41. The second phase was marked by the sudden onset of regular activity of the uterus; frequency rapidly increased and maximal values were reached around Day 49, after which frequency progressively decreased until around Day 66. The third phase was characterized by a steady state in genital tract activity until the peripartum period. The cervix and lesser curvature activity followed an almost similar development of regular activity episodes with only slight time differences.     

Plasma concentrations of testosterone,androstenedione, progesterone,oestradiol-17β, LH and FSH during the preovulatory period in the ewe

Seven mature ewes were synchronized to oestrus by two injections of 125 μg Cloprostenol given 9 days apart. Blood samples, collected for 72 h at 4-h intervals beginning 16 h after the second Cloprostenol injection, were assayed for testosterone, androstenedione, progesterone, oestradiol-17β, LH and FSH. For the testosterone measurements, two radioimmunoassays, using two different antisera, were validated and used. A typical pattern of release was observed for progesterone, oestradiol-17β, LH and FSH, with a preovulatory gonadotrophin peak recorded 16.1 ± 2.1 h after the observed oestrous behaviour. In two of the experimental animals, an extra oestradiol peak was recorded before the usual preovulatory rise. The changes in the concentrations of testosterone and androstenedione during the same period were not synchronous. The levels of the two androgens fluctuated considerably with occasional peaks of 150–250 pg/ml and even 900–1400 pg/ml. Although a tendency towards an increase in the levels of both androgens was observed during the period of oestrous behaviour, the individual variations were significant.     

Induction of male behaviour in ovariectomized ewes and ovariectomized-androgenized ewes chronically implanted with oestradiol-17β or testosterone

Normal mature ewes and ewes that had been androgenized with testosterone (T) between days 30–80 or 50–100 of fetal life were ovariectomized and given 100 mg implants of either oestradiol-17β (E) or T. The T implants caused a sustained elevation in plasma T levels but the E implants did not produce stable plasma levels of E. The implants were weighed on removal from the ewes and daily release rates for E and T were 14.4 ± 5.8 μg/kg/day and 24.2 ± 5.3 μg/kg/day respectively.The implants of E induced oestrous behaviour in both the non-androgenized and the androgenized ewes, some of these animals remaining in oestrus for up to 11 days. The ewes also began to mount each other after 1–9 days of treatments; the androgenized ewes also showed male-like aggressive behaviour whereas the non-androgenized ewes did not.The T implants induced oestrous behaviour in both androgenized and non-androgenized ewes. However, the non-androgenized ewes never mounted other ewes, nor did they show aggressive behaviour, whereas the androgenized ewes did.Prenatal androgenization clearly alters the ability of a ewe to respond to exogenous steroids by increasing its propensity to show masculine behaviour. Nevertheless, non-androgenized ewes may also show masculine behaviour during chronic steroid treatment.