Fertility studies in young and mature merino ewes 2. Sperm transport

Samples of cervical mucus were collected from young and mature ewes at different stages of oestrus. The migration rate of spermatozoa (time taken to travel 410 mm) was not different in samples of mucus from young and mature ewes. Spermatozoa were unable to penetrate the mucus in 7 27 samples, five of which were collected about 16 hours after onset of oestrus. Migration rate in samples collected between 10 and 16 hours after onset of oestrus was significantly slower than in samples collected in the first five hours of oestrus. The transport of spermatozoa in young and mature ewes was also studied by observing distribution of spermatozoa in the female genital tract 26 to 27 hours after hand-mating. There were no significant differences between young and mature ewes either in mean sperm count or in variation of mean sperm numbers in all sections of the tract.     

Cervical mucus and oestrous behaviour responses to oestrogens in sheep: Progesterone-oestrogen relationships and role of progesterone pre-treatment

The final dose of progesterone (5, 10, 20 mg) and time to oestrogen injection relative to the final dose of progesterone (24–72 h) had no significant effect on the production of cervical mucus measured 24 h after the injection of 30 μg oestradiol benzoate (ODB). However, there were significant effects on the behavioural oestrous responses (time from injection of oestrogen to onset of oestrus and duration of oestrus). Time to onset of oestrus increased from 18 to 27.8 h with increasing dose of progesterone (P < 0.001) and decreased from 24.8 to 20 h with increasing time to oestrogen injection (P < 0.05). Conversely, the duration of oestrus decreased from 36.2 to 23.8 h with increasing dose of progesterone (P < 0.001) and increased from 29 to 39 h with increasing time to oestrogen injection (P < 0.01).Ovariectomized ewes became refractory to ODB as measured by the cervical mucus response after the fifth sequential daily injection of 20 μg oestradiol benzoate. Progesterone priming was not required to restore subsequent sensitivity to oestrogen treatment. However, there was a positive linear relationship between length of recovery period and level of response to subsequent treatment.It was concluded that: (1) progesterone pre-treatment or priming is not necessary in the cervical mucus bioassay in ovariectomized ewes; and (2) a period of 8–16 days is needed between assays for normal sensitivity to be regained.     

Coincident increases in oxytocin receptor expression and EMG responsiveness to oxytocin in the ovine cervix at oestrus

This study has localised oxytocin receptor (OTR) mRNA expression within the cervix of non-pregnant ewes and related this to changes in the sensitivity of the cervical musculature to administered oxytocin (OT) during the oestrous cycle by recording electromyographic (EMG) activity. Cervices were collected from 34 ewes at specified time points throughout the cycle. OTR mRNA expression was localised by in situ hybridisation and results were expressed as optical density measurements from autoradiographs in each of four different cervical regions. EMG recordings were made for up to 8 h per day from four non-pregnant ewes undergoing seasonal oestrous cycles between Days −3 and +3 relative to oestrus (Day 0). The highest concentrations of OTR mRNA were detectable within the luminal epithelium (LE) of the cervix, with values increasing from Day 15 of the cycle, peaking during the follicular phase (P<0.001, compared to the mid-luteal phase) and returning to basal by Day 2. There was a small but significant increase in OTR mRNA hybridisation (above basal/luteal phase values) within the stromal cells (STR) adjacent to the lumen (P<0.05) during the same time period, but no differences from basal values were detectable in the dense collagenous annular ring or in tissue superficial to this. Analysis of pooled EMG activity recorded daily from the cervix indicated that endogenous contractile activity was higher on Day 0 than on the Days +1 (P<0.05), −2, +2 and +3 (P<0.001). The response to bolus intravenous (i.v.) injections of 25 mU OT (25 mU) varied with day of the cycle. This dose produced a measurable and significant response on Days 0 (P<0.001) and +1 (P<0.001), but not on any of the other days, indicating that the sensitivity of the cervical musculature to OT peaked on these days. These data show that the cervix is highly responsive to OT at oestrus. This coincides with an increase in OTR mRNA expression in the luminal epithelial cells, suggesting the likely production of an intermediary messenger between the epithelial and smooth muscle cells.     

Changed control of cervical secretion from infertile ewes previously exposed to oestrogenic clover pasture

The amount of cervical mucus recovered from control ovariectomized ewes increased with increasing doses of oestradiol benzoate (OB), while the maximum Spinnbarkeit of mucus occurred at an intermediate dose of OB. Neither the amount nor the Spinnbarkeit of mucus varied with the dose of OB in ewes with permanent infertility caused by grazing oestrogenic pasture (clover-affected ewes). Furthermore, the increase in Spinnbarkeit of cervical mucus seen in normal ewes treated over a 3-day period with OB or with implants containing oestradiol did not occur in affected ewes. In control ewes treated repeatedly with OB, production of mucus declined within 5 days, but no change in secretion was detectable in clover-affected ewes. Therefore, neither the amount nor the duration of oestrogenic stimulation affected the cervical mucus in ewes with clover disease. Affected ewes produced more mucus than did controls in the absence of oestrogenic stimulation. It is concluded that the relatively normal volume of mucus in affected ewes treated with OB results largely from autonomous production. The Spinnbarkeit does not increase in these ewes because the ability of the cervix to respond to oestrogen is impaired.     

Assessing the usefulness of prostaglandin E2 (Cervidil) for transcervical artificial insemination in ewes

The underlying theme of this study involved the evaluation of the dilatory effects of prostaglandin E₂ on the ovine cervix and thus the assessment of its potential applicability to transcervical artificial insemination (TCAI) in ewes. A novel method of prostaglandin E₂ administration (controlled slow-release vaginal inserts) was examined, and the practical implications of this approach including cervical penetrability and posttreatment pregnancy rates were evaluated. The Guelph method of TCAI was performed during the seasonal anestrus (n = 40) and the breeding season (n = 40) on multiparous Rideau Arcott × Polled Dorset ewes, with or without the pretreatment with Cervidil (for a duration of 12 hours or 24 hours before TCAI). Cervical penetration rates averaged 82.5% (66 of 80), and they varied neither (P > 0.05) between the two seasons nor between Cervidil-treated ewes and their respective controls. Cervidil priming significantly reduced the total time required for TCAI during the breeding season in comparison with controls (54 vs. 98 seconds), especially after the 24-hour exposure (38 vs. 108 seconds). The time taken to traverse the uterine cervix was negatively correlated (P < 0.05) with the breed (percentage of Rideau Arcott genotype) and lifetime lamb production in seasonally anestrous ewes. Four out of 36 (11%) successfully penetrated ewes in the breeding season (three ewes allocated to the 12-hour control group and one ewe that had received Cervidil for 12 hours) became pregnant and carried the lambs to term. Vaginal mucus impedance at TCAI was significantly and positively correlated with the total time required to complete the procedure in cyclic ewes, and the negative correlation between vaginal mucus impedance and total time values at the time of controlled intravaginal drug release device removal approached to significance in anestrous ewes. The present results indicate a moderate benefit of using Cervidil for inducing cervical dilation before TCAI in ewes, mainly in the breeding season. The specific reason(s) for impaired fertility after the TCAI using frozen–thawed ram semen remains to be elucidated.     

Effect of site of deposition on the fertility of sheep inseminated with frozen-thawed semen

The widespread use of artificial insemination (AI) in sheep is currently prevented due to the lack of a cost effective insemination technique utilising frozen-thawed semen. The objective of the present study was to determine if the deposition of frozen-thawed semen in the vaginal fornix would result in a pregnancy rate comparable to that achieved following cervical insemination. Multiparous ewes of various breeds were synchronised and inseminated into either the vaginal fornix (n=78) or the cervix (n=79), at 57 h post sponge removal, with frozen-thawed semen. Information on mucus secretion and the depth to which it was possible to penetrate the cervix at insemination (cervically inseminated ewes only) was recorded at the time of AI. Pregnancy rate was subsequently determined either by return to service (oestrus) or after slaughter 30 days post insemination. Insemination site did not significantly influence pregnancy rate using frozen-thawed semen (36.2% compared to 27.6% for cervical and vaginal fornix insemination, respectively; P=0.26). Whilst depth of cervical penetration was positively associated with pregnancy rate (P<0.05), this association needs to be interpreted with caution as none of the ewes where the cervix could not be penetrated (score=0) was pregnant. In conclusion, pregnancy rate following insemination of frozen-thawed semen into the vaginal fornix was within 10% points of that obtained following cervical AI of frozen-thawed semen. As insemination into the vaginal fornix is technically easier than cervical insemination, it may be more practical for use in large scale applications.     

Persistent infertility in ewes after prolonged exposure to oestradiol-17 beta

Merino ewes were treated with implants which released 300 micrograms oestradiol-17 beta per day or 5 mg progesterone per day, or both, for 9 months (Months 1-9), and after an 11-month intermission were treated again for 6 months (Months 20-26). Ewes were run with rams at Months 16, 28 and 40. Fertility was not affected by the first exposure period, but the second exposure to oestradiol reduced the fertility of ewes at both subsequent mating periods. Affected ewes returned to service more frequently (P less than 0.01) and were less likely to conceive (P less than 0.05). After mating, a normal population of spermatozoa was established in the caudal cervix, but transport through the cervix was impaired in affected ewes and there were fewer spermatozoa (P less than 0.01) in the cranial cervix. In affected ewes, the spinnbarkeit of cervical mucus was reduced (P less than 0.05), and the histological appearance of the cervix changed, looking like that of the uterus. Treatment with progesterone did not affect fertility, cervical mucus or sperm transport, but diminished the histological abnormalities produced by oestradiol (P less than 0.05). These results show that oestradiol-17 beta given after puberty can cause the same kind of permanent sexual transdifferentiation that is produced by the oestrogenic isoflavones in ewes with clover disease. The results suggest that this change may require more than a single exposure to oestrogen.     

Continuous infusion of oxytocin prevents induction of uterine oxytocin receptor and blocks luteal regression in cyclic ewes

Continuous intravenous infusion of oxytocin (3 micrograms/h) between Days 13 and 21 after oestrus delayed return to oestrus by 7 days (length of cycle 23.3 +/- 0.6 days compared to 16.6 +/- 0.2 days in control ewes). At a lower infusion rate (0.3 micrograms/h) oxytocin delayed luteolysis in only 2 of 5 ewes. Treatment from Day 14, when luteolysis had already begun, was ineffective. Delay of luteal regression by oxytocin had no effect on the length of subsequent cycles. Measurement of circulating progesterone concentrations and luteal weight showed that prolongation of the oestrous cycle was due to prevention of luteal regression. Luteal regression and behavioural oestrus were induced during continuous oxytocin administration begun on Day 13 when cloprostenol was given on Day 15 (mean cycle length, 17.3 +/- 0.21 days). Continuous oxytocin infusion from Day 13 blocked the rise in uterine oxytocin receptor concentrations which normally precedes oestrus. Mean receptor concentrations in caruncular and intercaruncular endometrium and in myometrium were 76, 36 and 9 fmol/mg protein on Day 17 in ewes receiving continuous oxytocin (3 micrograms/h); in control ewes these values were 675, 638 and 130 fmol/mg protein respectively at oestrus. Receptor concentrations on the day of oestrus in ewes receiving oxytocin and cloprostenol were not significantly different from those in control ewes (649, 852, and 109 fmol/mg protein respectively). Since cloprostenol, a PGF-2 alpha analogue, overcame the antiluteolytic action of oxytocin, it is suggested that continuous oxytocin treatment may inhibit uterine production of PGF-2 alpha, possibly by down regulating the uterine oxytocin receptor.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effects of the prostaglandin E analogue Misoprostol and follicle-stimulating hormone on cervical penetrability in ewes during the peri-ovulatory period

Two experiments in parous Welsh Mountain ewes determined the pattern of natural cervical relaxation over the peri-ovulatory period and investigated FSH and Misoprostol as cervical relaxants to facilitate transcervical passage of an insemination pipette into the uterine cavity. Following synchronisation of oestrus using progestagen sponges and PMSG (500 IU) the depth of cervical penetration was determined using a modified cattle insemination pipette as a measuring device. Penetration of the cervix was least at the time of sponge removal and increased to a maximum at 72 h after sponge removal and then declined. Intra-cervical administrations of either ovine FSH (Ovagen; 2mg) or Misoprostol (1mg; a Prostaglandin E(1) analogue) facilitated cervical penetration. Ovagen given 24h after sponge removal allowed transcervical intrauterine penetration in 100% of ewes at 54 and 60 h after sponge removal while Misoprostol given 48 h after sponge removal allowed trans-cervical penetration in 100% of ewes at 54 h. A combination of Ovagen and Misoprostol was as effective but not more so than Ovagen or Misoprostol alone. These results show that there is natural relaxation of the cervix at oestrus and that maximum relaxation occurs 72 h after sponge removal, which is too late for the correct timing of insemination. The intra-cervical administration of FSH or Misoprostol enhanced relaxation of the cervix and both were able to relax the cervix to allow intrauterine penetration 54 h after sponge removal, the optimum time for insemination. The results also show that FSH is biologically active after intracervical, topical application.     

Ovulation, fertilization and lambing rates, and peripheral progesterone concentrations, in ewes inseminated at a natural oestrus during November or February.

The objective of this study was to determine the relative importance of seasonal changes in ovulation rate, fertilization rate and embryo survival as the cause of reduced lambing rates in ewes mated in February compared with those mated in November. The study was conducted at 57 degrees N using mature Mule ewes and Suffolk rams. Sixty ewes were allocated equally to five groups: unbred (UB) or mated at a natural oestrus during November (N) or February (F) by natural (N) or cervical artificial (A) insemination. Groups were maintained separately at pasture supplemented with hay. A raddled vasectomized or non-vasectomized ram was present with UB, NN and NA groups from 26 October 1995 to 1 January 1996 and with UB, FN and FA groups from 25 January 1996 to 31 March 1996. Ewes marked by the ram were recorded twice a day, and those in groups NN, NA, FN and FA were inseminated at their second behavioural oestrus. For all ewes, blood samples were obtained once a day from introduction of the vasectomized rams until 30 days after mating (groups NN, NA, FN and FA) or 20 days after the first oestrus (group UB), and ovulation rate was measured by laparoscopy 7 days after the first oestrus. For ewes in groups NN, NA, FN and FA, ovulation rate was measured again after the second oestrus and ova were recovered from six ewes per group for assessment of fertilization before autotransfer. Pregnancy and lambing rates were recorded at term. Mean (+/- SE) dates of the first recorded oestrus for ewes in groups NN, NA and UB, and FN, FA and UB were 4 +/- 1.1 November and 4 +/- 0.9 February, respectively, and intervals between the first and second oestrus were 16 +/- 0.2 and 17 +/- 0.3 days (P < 0.01), respectively. Ovulation rates were 2.6 +/- 0.08 and 2.0 +/- 0.05 (P < 0.001), and peripheral progesterone concentrations during the luteal phase were 8.5 +/- 0.25 and 7.6 +/- 0.31 ng ml-1 (P < 0.05), for November and February, respectively. The difference in peripheral progesterone concentration was not solely attributable to the difference in ovulation rate. There was no significant effect of month or method of insemination, or of embryo recovery and autotransfer procedures on pregnancy rates and the proportion of ewes that became pregnant were NN 0.92, NA 0.83, FN 0.67 and FA 0.75. For ewes undergoing embryo recovery and autotransfer, ova recovered per corpus luteum were 1.00, 0.93, 1.00 and 0.92, fertilized ova per ovum recovered were 0.69, 0.92, 1.00 and 0.83, and lambs born per corpus luteum were 0.62, 0.79, 0.78 and 0.58 for NN, NA, FN and FA groups, respectively. There were no significant seasonal effects on fertilization rate or embryo survival. It is concluded that a seasonal decline in ovulation rate is the primary cause of reduced lambing rates in ewes mated in February compared with those mated in November. Pregnancy rates were high after mating in both periods and were not enhanced by the use of cervical insemination.     

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.     

Fertility and sperm transport in Merino ewes at the first oestrus following embryonic death.

The embryos of ewes were killed with colchicine on Day 17 of gestation and the ewes were mated at the subsequent oestrus. Fertility was reduced at this mating, and fewer spermatozoa were found in the uterus and oviducts than in control animals. The total number of spermatozoa in the cervix and their distribution between the lumen and walls of the cervix were not altered, but the linear distribution along the cervical walls was changed. The density of the reamining spermatozoa in the control animals after flushing the cervix showed a progressive decrease from the posterior to the anterior segments. This did not occur in the untreated ewes. It seems likely that impaired sperm transport contributed to the lowered fertility.     

The evaluation of a laparoscopic insemination technique in ewes

Following synchronisation of oestrus using FGA and PMSG, ewes were inseminated by either the conventional cervical (CC) method or directly into the uterus by laparoscopy (LI). The CC method was carried out either at 48 and 60 hours following progestagen withdrawal with 480 x 10(6) spermatozoa per inseminate or once only at 56 hours with 600 x 10(6) spermatozoa. The laparoscopic method was performed at 52 hr using 48 x 10(6) spermatozoa per ewe. In the first two trials eggs were recovered at laparotomy. The egg recovery rate was significantly lower (P<0.05) for those ewes which had been inseminated by the LI method (74%) compared with those inseminated by the CC method (85%); fertilization rates were not significantly different (92% and 89% respectively). In the third trial 20 ewes were bled to determine their periovulatory LH concentrations and the timing of peak LH concentrations correlated with the outcome of each insemination. Ewes inseminated using laparoscopy did not conceive when their LH surge occurred >58 hr after progestagen withdrawal. In this and in the final experiment, the combined pregnancy rates and litter sizes (assessed radiographically) were 67% (n = 51) and 2.21 (n = 34) for the CC method and 75% (n = 48) and 1.97 (n = 36) for the LI method (P>0.05).     

A study of oestrus and fecundity in a laboratory colony of Mouse opossums (Marmosa robinsoni)

Twenty wild caught Mouse opossums Marmosa robinsoni Bangs 1898 formed the basis of a laboratory colony which was maintained at the Brookfield Zoo during 1971–3. Approximately 250 young including 3rd-generation descendants were produced.
M. robinsoni is polyoestrous with a mean cycle length of 25.5±0.5 days. Vaginal oestrus, which normally lasts three days, is accompanied by a pronounced cycle of cornification and mucification. During this period large numbers of desquamated epithelial cells and mucus are found in the urine. Females are sexually receptive 24 hours before cornified cells first appear in the urine and receptivity lasts two to four days. Fertile copulation takes place throughout this period. Ovulation is spontaneous and occurs late in oestrus. The mean number of corpora lutea recorded was 19.8±0.9 ( n =23).
Anovular cycles, associated with atresia of mature Graafian follicles, were found in approximately 25% of the females. The sterile cycles, which had a mean length of 15.6±0.7 days, were not always accompanied by a behavioural oestrus. Observations on the behaviour of introduced pairs provided direct evidence of sexual incompatibility. Sexual incompatibility was found to have been responsible for at least 10% of the unsuccessful pairings recorded during the study.
There were no indications in the Brookfield colony of the decline in productivity among laboratory-bred animals reported in other Marmosa colonies. Litter-sizes and litter-rates in the F₁ and F₂ generations did not differ from those in the wild-caught generation. However, the fact that only one out of three pairings produced young suggests that productivity in the colony was lower than in the wild. The cause of this low productivity was not established.     

Artificial insemination of ewes with frozen-thawed semen at a synchronized oestrus. 1. Effect of time of onset of oestrus, ovulation and insemination on fertility

In three experiments, the onset of oestrus, time of ovulation and lambing after intrauterine insemination with frozen-thawed semen were examined following synchronisation of oestrus using intravaginal progestagen-impregnated sponges (inserted for 12 days) and an injection of PMSG at sponge removal.

The number (and percentage) of ewes detected in oestrus 12, 24, 36, 48, 60 and 72 h after sponge removal was 1 (0.3), 2 (0.6), 17 (5.2), 120 (36.7), 65 (20.0) and 10 (3.1) respectively. One hundred and twelve ewes (34.3%) remained unmarked. Egg fertilisation rates were not different between ewes irrespective of time of onset of oestrus or whether or not ewes were marked.

The median time of ovulation with respect to sponge removal (with 95% fiducial limits) for ewes joined with vasectomised rams (10:1) at spronge removal (teased ewes) was 55.8 h (54.61–57.09) and for unteased ewes 59.7 h (58.27–61.12).

In the third experiment, a total of 394 ewes were inseminated by laparoscopy with frozen-thawed semen. The percentage of ewes lambing and lambs born per ewe inseminated, and number of lambs born per ewe lambing for inseminations 48, 60, 72 and 78 h after sponge removal were 45.9, 57.7 and 1.25; 55.1, 72.0 and 1.31; 57.4, 80.9 and 1.41; and 39.3, 60.7 and 1.54, and for 59 control ewes receiving fresh semen by cervical insemination 47.5, 69.5 and 1.46 respectively. The lambing data after insemination with frozen semen was not different to that of the controls. The percentage of ewes lambing and lambs born per ewe inseminated increased with time of insemination at 48, 60 and 72 h (linear, P < 0.01) but was lower for inseminations at 78 h after sponge removal. Number of lambs born per ewe lambing increased with time of insemination after sponge removal (linear, P < 0.05).     

Effect of nutritional flushing on the productivity of Finnish Landrace ewes

In the autumn (October–December), Finnish Landrace ewes aged 1 year (yearling), 4–5 years (mature) and 7–8 years (older) were fed forages with no supplemented (control) or supplemented daily with 150 g (diet 150) or 300 g (diet 300) of concentrates per ewe in order to determine the effect of energy supply to initiate flushing on ewe productivity. Concurrently, in the summer (August–September), yearling and mature ewes were fed 0, 300 or 450 g of concentrates daily for 17 or 34 days. Concentrate supplement prior to and during mating had little effect on the live weight, however mature ewes gained weight in the summer. In the autumn, increasing proportion of concentrate in the diet reduced the length of the mating period in the yearling ewes (P=0.03) however, diet combination had no significant effect of age on conception. In August and early September, flushing did not hasten the onset of estrus and only 25% of the young ewes and 83% of the adult ewes were mated successfully. The flushing diet had no effect on the prolificacy in yearling and older ewes. In contrast, prolificacy and number of lambs weaned increased (P<0.05) for mature ewes on diet 150. In the autumn, there was a significant influence of flushing on the litter weight at birth and live weight gain from birth to weaning for older ewes however, diet 300 and the control were similar. Although litter weight at birth and weaning tended to be higher in the summer for ewes supplemented with concentrate for 34 days, the number of ewes required for a significant influence due to diet and length of flushing combination were not adequate. It was concluded that little is to be gained in lamb production by flushing ewes of the Finnish Landrace breed.     

Plasma progesterone concentrations, ovarian and endocrinological response and sperm transport in ewes with synchronized oestrus

Two experiments involving 24 and 54 Australian Merino ewes were conducted in which the establishment of a cervical population of spermatozoa and several endocrinological events were studied after several regimens for the synchronization of oestrus. Intravaginal sponges impregnated with 500 mg (Exp. 1) or 200, 400 or 600 mg (Exp. 2) progesterone resulted in the maintenance of plasma progesterone concentrations of 1.5-4.9 ng/ml over a 12-day insertion period compared with 1.9-6.9 ng/ml during dioestrus in control ewes. In Exp. 1 basal concentrations of less than or equal to 0.25 ng/ml plasma were attained by 4 h after sponge withdrawal and this decline was much more rapid than in normal luteolysis. This was associated with fewer spermatozoa recovered from the cervix 2 h after insemination, and PMSG had no significant effect. In Exp. 2 injection of a supplementary dose of progesterone at sponge withdrawal resulted in a rapid increase in plasma progesterone concentrations followed by an equally rapid decrease and an attenuation of the rise in plasma oestradiol-17 beta, the LH surge, and the onset of oestrus. The numbers of spermatozoa recovered 4 h after insemination were not increased, and PMSG had no significant effect. Two factors were significant, namely the dose of progesterone in the sponge (600 mg greater than 400 or 200 mg, P less than 0.05) and stage of oestrus when inseminated (mid- or late oestrus greater than early). The data demonstrated that an adequate dose of progesterone/progestagen incorporated into intravaginal sponges and accurate timing of insemination relative to the LH surge are the most important factors involved in penetration of the cervix by spermatozoa.     

Oestrogen and progesterone concentrations in plasma and oviductal tissue of ewes exhibiting a natural or induced oestrus

Synchronisation of oestrus in Karagouniki ewes by administration of the standard dose of progesterone results in lower fertility than observed when these ewes ovulate naturally. This suggests that the optimum dose of progesterone may be breed dependent. The exogenous progesterone may perturb the concentrations of oestradiol-17beta and progesterone in blood plasma and the oviductal wall. This possibility was investigated using Karagouniki ewes allocated at random to three treatments (n=4 per treatment). Ewes were allowed to exhibit natural oestrus (N) or oestrus was synchronised by administration of 250 mg (LP) or 375 mg (HP) progesterone (subcutaneous implants) followed by PMSG at 8 mg/kg live weight i.m. 14 days later. Oestrus was observed using teaser rams. Blood samples were collected for plasma oestradiol-17beta and progesterone assay from the onset to the end of oestrus at 2 h intervals. The uterus of each ewe was recovered at the end of oestrus and samples of the oviductal wall were taken from both oviducts and prepared, separately, for progesterone and oestradiol-17beta assay. Statistical analysis was performed using univariate analysis of variance. Plasma oestradiol-17beta concentrations from the onset to the end of oestrus were highest for N ewes and lowest for HP ewes with the values for LP ewes occupying an intermediate position. The differences were significant (P<0.05) between HP and the other two treatments from 4 to 12 h after the onset of oestrus and then between all treatments until the end of oestrus. Plasma progesterone levels were similar and fairly constant from the onset to the end of oestrus for N and LP. The plasma progesterone levels for HP were significantly (P<0.05) higher than for the other two treatments throughout oestrus. In oviductal wall samples, the oestradiol-17beta concentration was significantly (P<0.05) higher for N ewes than for synchronised ewes and the levels were similar for LP and HP ewes. The concentration of oestradiol-17beta differed (P<0.05) between right and left oviducts for N ewes but not for ewes of either of the synchronised oestrus treatments. Progesterone concentrations in oviductal wall samples were highest (P<0.05) for HP ewes and the values for N and LP ewes were similar. The concentration of progesterone did not differ between right and left oviductal wall samples within treatments. It was concluded that the higher dose of exogenous progesterone perturbed the levels of oestradiol-17beta and progesterone in blood plasma and the oviductal wall, and this could explain the lower levels of fertility (relative to naturally occurring oestrus) observed when this protocol is used for Karagouniki ewes in practice.     

Oestrous cycle dynamics in peri-pubertal and mature Javanese thin-tail sheep

Duration of oestrus, time of ovulation and hormone profiles for progesterone and LH in prepubertal, pubertal and mature Javanese thin-tail sheep were studied at synchronized oestrus following progestagen-PMSG treatment and at the first natural oestrus after synchronization.The ewe lambs responded to progestagen-PMSG treatment by showing earlier onset of oestrus and an earlier and higher peak of LH concentration than mature ewes. For pre-pubertal, pubertal and mature ewes the mean LH peaks were 49.9, 43.9 and 37.9 ng/ml (P>0.05) at mean intervals of 7.5, 8.4 and 16.5 h (P < 0.05), respectively, after onset of oestrus. Duration of oestrus was 41.2 h in pubertal lambs and averaged 37.5 h in the other two groups (P>0.05). Except in one mature ewe, ovulation occurred between 24 and 36 h after onset of oestrus and the majority ovulated at around the end of oestrus. The corpora lutea developed normally, as indicated by plasma-progesterone changes. The patterns of plasma-progesterone changes were similar in all three groups, though the concentrations were lower in the ewe lambs.At the first natural oestrus after synchronization, mature ewes showed longer (P>0.05) oestrus (31.5 vs. 24.3 h), longer time interval from onset of oestrus to the LH peak (16.0 vs. 12.0 h) and from the LH peak to ovulation (21.0 vs. 19.6 h) than peri-pubertal lambs. Six of eight pre-pubertal lambs did not ovulate at their first natural oestrus, resulting in a conception rate of 11% for that group, while in pubertal lambs and mature ewes conception rates were 70% and 100%, respectively.     

Influence of haemoglobin type and selenium status on peripheral plasma progesterone and corticosteroid concentration in ewes grazing oestrogenic pastures.

Peripheral plasma progesterone and corticosteroid concentrations were studied in ewes grazing oestrogenic pastures during oestrus, pregnancy and parturition in 1974 and during pregnancy in 1975. Haemoglobin (Hb) type A ewes had significantly higher mean corticosteroid concentrations than HbB ewes in both years. In 1974 the concentration of progesterone did not differ significantly between Hb types whilst in 1975 the concentration was significantly (P less than 0.001) higher in HbA ewes. The mean corticosteroid concentrations were higher in selenium-supplemented ewes of both Hb types than in unsupplemented ewes in 1974 but lower in 1975. Selenium treatment did not significantly influence the progesterone concentration in ewes of both Hb types in 1974 but significantly (P less than 0.01) increased in 1975. It is concluded that the higher concentration of plasma corticosteroids or factors regulating the relationship between corticosteroids and progesterone could be responsible for the higher estimates of embryonic mortality reported in HbA ewes.