Effects of breed, ovarian steroids and season on the pulsatile secretion of LH in ovariectomized ewes.

The effects of season and of oestradiol and progesterone on the tonic secretion of LH were studied in ovariectomized Merino and Suffolk ewes, two breeds which differ markedly in the seasonal pattern of their reproductive activity. In the absence of exogenous steroids, the frequency of LH pulses was lower and the amplitude of the pulses was higher in anoestrus than in the breeding season for Merino and Suffolk ewes 30 days after ovariectomy. In long-term (190 days) ovariectomized ewes, this seasonal change in LH secretion was observed in Suffolk ewes only. During seasonal anoestrus, treatment of ewes with subcutaneous oestradiol-17 beta implants (3, 6 or 12 mm in length) decreased the frequency of LH pulses in a dose-dependent manner, with Suffolk ewes being far more sensitive to the inhibitory effects of oestradiol than Merino ewes. The lowest dose of oestradiol (3 mm) had no effect on the secretion of LH in Merino ewes, but reduced secretion in Suffolk ewes. Treatment of ewes with the highest dose of oestradiol (12 mm) completely abolished LH pulses in Suffolk ewes, whereas infrequent pulses remained evident in Merino ewes. During the breeding season, oestradiol alone had no effect on the pulsatile release of LH in either breed, but in combination with progesterone there was a significant reduction in LH pulse frequency. Progesterone effectively decreased LH secretion in both breeds in both seasons. It was concluded that differences between breeds in the 'depth' of anoestrus could be related to differences in the sensitivity of the hypothalamus to both negative feedback by oestradiol and the direct effects of photoperiod.

Importance of non-olfactory ram stimuli in mediating ram-induced ovulation in the ewe.

In Exp. 1, 4 groups of 50 recently weaned ewes were exposed to various degrees of contact with rams for 65 days, followed by exposure to novel rams for 4 days. Ovarian activity in the ewes was determined by laparoscopy on Days 29, 65 and 69 of treatment. There were no treatment differences in the percentage of ewes ovulating on Day 4 whereas by Day 29 more ewes in clear fenceline and full ram contact were ovulating compared to controls (P less than 0.05, P less than 0.001). After 65 days ovarian activity was significant only in those ewes in full contact with rams (P less than 0.001). Between 89 and 95% of ewes remaining anovulatory after 65 days ovulated after 4 days of full contact with novel rams. In Exp. 2, 4 groups of about 30 anovulatory ewes were exposed to various degrees of contact with rams for 5 days. Ovarian activity was assessed before and after treatment by laparoscopy. After 5 days, more ewes were ovulating in response to full ram contact than in any other treatment (P less than 0.05) and more ewes in fenceline contact with rams or with rams plus ewes were ovulating than in the isolated control treatment (P less than 0.01). In Exp. 3, 6 groups of about 40 anovulatory ewes were exposed to face masks with and without rams' wool and/or various degrees of contact with rams for 5 days. More ewes were ovulating after 5 days in the group in full physical contact with rams than in any other group (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

The number of spermatozoa, the number of ovulations per ewe, and immunization against androstenedione affect fertility and prolificacy of sheep.

Ewes that were untreated, fed lupins or fed lupins and immunized against androstenedione were artificially inseminated. The percentage of ewes pregnant at 36-45 days after insemination (fertility) was 8% higher in ewes that had more than one ovulation than in those that had only one ovulation. Maximum fertility was achieved with 50 x 10(6) spermatozoa and this did not vary with the number of ovulations that ewes had. Among the pregnant, twin-ovulating ewes, embryo survival increased as the number of spermatozoa inseminated increased from 25 x 10(6) to 400 x 10(6). Immunization of ewes against androstenedione increased ovulation rate but reduced fertility, and reduced embryo survival among twin-ovulation ewes.

Ovulation in the merino ewe in the breeding and anoestrous seasons.

The pattern of ovulation of Merino ewes was studied by repeated laparoscopy each 14 days in the anoestrous (n = 97) and breeding (n = 87) seasons. In the anoestrous season the proportion of ewes ovulating did not decrease below 11%, 42% of ewes never ovulated and the remainder fluctuated between the two states. On 20 occasions a clear anovulatory period was interrupted by an isolated spontaneous ovulation. In the breeding season the overall mean proportion of ewes with corpora lutea or albicantia at laparoscopy was 87%, 54% of ewes ovulated regularly throughout while in another 31% absence of corpora lutea or albicantia coincided with the follicular phase of an oestrous cycle as evidenced by an appropriately aged corpora lutea at the next laparoscopy. Of the remaining 15% of the flock 3% had anovulatory periods greater than 14 days while the remainder experienced irregular ovulatory cycles--the majority due to short periods of anovulation but some ewes retained corpora lutea for longer than 14 days while others ovulated twice between successive laparoscopies.

Effect of immunization against progesterone on oestrus, cycle length, ovulation rate, luteal regression and LH secretion in the ewe.

The effects of active immunization against progesterone on reproductive activity were studied in Merino ewes. Immunization against progesterone caused a shortening (P less than 0.01) of the interval between ovulations from 17-18 days (controls) to between 6 and 10 days (immunized group); this was associated with a corresponding reduction in the interval between LH surges. The immunized ewes also had higher (P less than 0.05) ovulation rates (1.72) than controls (1.25) and exhibited a reduced (P less than 0.01) incidence of oestrus (26% v. 95%). Many immunized ewes continued to ovulate despite the persistence of corpora lutea from earlier ovulations which led to an accumulation on the ovaries of many corpora lutea of different ages. The frequency of LH pulses in ewes immunized against progesterone (1.8 +/- 0.2 pulses/4 h) was significantly (P less than 0.001) higher than that of control ewes (0.3 +/- 0.1 pulses/4 h). This study highlights the importance of progesterone in the control of oestrus, ovulation, ovulation rate, luteal regression and the secretion of LH in the ewe.

Corpora lutea with a short life-span induced by rams in seasonally anovulatory ewes are prevented by progesterone delaying the preovulatory surge of LH.

Seasonally anovulatory Merino ewes isolated from rams were allocated to three treatments before the re-introduction of rams. Ten ewes received a single injection of progesterone (20 mg), 18 ewes received the injection of progesterone but had the ram-induced preovulatory surge of LH replaced by a series of injections of GnRH 24 h after the introduction of the rams, and 20 control ewes had no hormone treatment. Of the 48 ewes, 44 ovulated within 5 days of the introduction of rams and the treatments had no significant effect on the incidence of ovulation. The frequency of corpora lutea with a short life span (the interval between successive preovulatory surges of LH being 5.1 +/- 0.9 days) was 72% for control ewes and 58% for ewes treated with progesterone and GnRH, but such CL were prevented completely after the injection of progesterone alone (P less than 0.001). The injection of progesterone also delayed the preovulatory surge of LH (P less than 0.001). These results suggest that progesterone assures normality of corpora lutea by lengthening the period of gonadotrophin priming of follicles before ovulation.

Progesterone priming and age of ewe affect the life-span of corpora lutea induced in the seasonally anovulatory Merino ewe by the 'ram effect'.

Increasing doses of progesterone from 0 to 20 mg, given as a single i.m. injection, increased the proportion of corpora lutea that had a normal life-span when induced in ewes by the introduction of testosterone-treated wethers from 54% (19/35) to 100% (34/34). Injection of progesterone did not affect the induction of ovulation and 95% (130/136) of the anovulatory ewes ovulated. Nevertheless, a low proportion of ewes displayed oestrus between Days 16 and 26 after the introduction of testosterone-treated wethers (Exp. 1, 47%, 92/196; Exp. 2, 50%, 502/1000). Many of the ewes that did not display oestrus also failed to ovulate again (Exp. 1, 70%, 37/53). The proportions of anovulatory adult and maiden ewes that ovulated after the introduction of testosterone-treated wethers were not significantly different but significantly fewer maiden ewes were detected in oestrus.

Effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus.

The effects of progesterone on the responses of Merino ewes to the introduction of rams during anoestrus were investigated in two experiments. In the first experiment, the introduction of rams induced an increase in the levels of LH in entire ewes. The mean levels increased from 0.68 +/- 0.04 ng/ml (mean +/- s.e.m.) to 4.49 +/- 1.32 ng/ml within 20 min in ewes not treated with progesterone (n = 10). In ewes bearing progesterone implants that provided a peripheral concentration of about 1.5 ng progesterone per millilitre plasma, the LH response to the introduction of rams was not prevented, but was reduced in size so that the concentration was 1.38 +/- 0.15 ng/ml after 20 min (n = 5). Progesterone treatment begun either 2 days before or 6 h after the introduction of rams and maintained for 4 days prevented ovulation. In the second experiment ovariectomized ewes were used to investigate further the mechanism by which the ram evoked increases in tonic LH secretion. In ovariectomized ewes treated with oestradiol implants, the introduction of rams increased the frequency of the LH pulses and the basal level of LH. In the absence of oestradiol there was no significant change in pulse frequency but a small increase in basal levels. Progesterone again did not prevent but reduced the responses in ewes treated with oestradiol. It is suggested that following the withdrawal of progesterone treatment, the secretion of LH pulses in response to the ram effect would be dampened. This effect could be a component of the reported long delay between the introduction of rams and the preovulatory surge of LH in ewes treated with progesterone. Continued progesterone treatment prevented ovulation, probably by blocking positive feedback by oestradiol.

Effect of stress due to laparoscopy on plasma cortisol levels, the preovulatory surge of LH, and ovulation in the ewe.

The ovulation which is induced in seasonally anovular ewes by the introduction of rams was used as a model to examine the effects of the stress of repeated laparoscopy on the preovulatory surge of LH and ovulation. The number of ewes which experienced LH surges and ovulation was reduced (10/23 vs 1/20) when laparoscopy was repeated every four hours. Plasma cortisol levels were used as an indicator of stress, and were found to increase after laparoscopy: a single laparoscopy produced a transient rise, while repeated laparoscopy induced and maintained levels between 70 and 100 ng/ml.

Abnormal function of the corpus luteum in some ewes with phyto-oestrogenic infertility.

Ewes with permanent phyto-estrogenic infertility show oestrus less regularly than normal ewes, and the present study examines the extent to which this results from abnormal ovarian function. Forty-nine affected ewes and 53 controls were run with rams fitted with marking crayons and harnesses, and crayon marks were recorded and laparoscopy performed at weekly intervals for 3 weeks. Fewer affected ewes showed oestrus accompanied by ovulation (28 v. 49, P less than 0.001), and four of these affected ewes had a second ovulation during the experiment. More of the ovulations observed in affected ewes were unaccompanied by behavioural oestrus than in controls (8 out of 38 v. 2 out of 50; P less than 0.05). Six affected ewes had no corpus luteum or oestrus, and five of these had adhesions over the genitalia. Hydrops uteri in five other affected ewes was accompanied by prolonged maintenance of the corpus luteum. Some other abnormalities were also observed. In a second study, plasma progesterone concentrations were measured twice daily in 12 affected ewes which were run with rams. Five ewes had oestrous cycles of abnormal duration (two of more than 23 days, two of 21 days, and one of 11 days), and these were accompanied by plasma progesterone patterns different from those of the ewes with an oestrous cycle duration of 16-18 days. It is concluded that the irregular oestrous cycles in affected ewes are due mainly to abnormal life span and progesterone secretion by the corpus luteum, which in turn largely result from changes in the uterus.

Ovulation rate and oocyte numbers in ewes after prolonged exposure to oestrogenic pasture.

Ewes which had been exposed to oestrogenic clover for 3 years showed an elevated ovulation rate but no difference in numbers of primordial follicles compared with normal ewes.

Laparoscopic examination of the reproductive tracts of gilts using a special crate.

A crate for restraining gilts for routine laparoscopic examination of the reproductive tract is described, together with the techniques employed in anaesthesia and laparoscopy. The procedure is efficient, allowing two operators to examine 4-5 gilts per hour.