Ovarian responses and pregnancy rate with previously used intravaginal progesterone releasing devices for fixed-time artificial insemination in sheep.

The objective was to determine serum progesterone concentrations, ovarian responses, and pregnancy rate in sheep inseminated following a short-term protocol (6 days of treatment) with a previously used controlled internal drug release-G (CIDR-G) device. In experiment 1, 30 ewes were put on a short-term protocol using a CIDR-G of first use (new devices, N = 10), second use (previously used for 6 days, N = 10), or third use (previously used twice for 6 days, N = 10). All ewes were given prostaglandin F(2α) (10 mg dinoprost) and eCG (300 IU) im at device withdrawal. Mean serum progesterone concentrations were greater for ewes treated with new versus reused devices (P < 0.05), but there were no significant effects on ovarian follicular development, namely: proportion of ewes that reached ovulation/treated ewes (8/10, 9/10, and 10/10); day of emergence of the ovulatory follicle (2.9 ± 1.8, 1.8 ± 2.4, and 2.5 ± 1.1 days after CIDR-G insertion); and lifespan of the ovulatory follicle (5.4 ± 1.8, 6.5 ± 2.2, and 5.6 ± 1.0 days) for devices of first, second, and third use, respectively. The proportion of ewes that ovulated from a new follicle emerging after CIDR-G insertion was 100% (10/10) with new devices, and 77.8% (7/9) and 80% (8/10) with devices used two and three times, respectively (P = not significant). In experiment 2, ewes (N = 319) received the short-term protocol as in experiment 1, using CIDR-G of first or third use (N = 158 and N = 161, respectively) for 6 days, with prostaglandin F(2α) and eCG given im at device withdrawal. Intrauterine (laparoscopy) fixed-time AI with fresh semen (100 × 10(6) sperm) was done 52 to 57 hours after device withdrawal. Pregnancy rates were 80.4% (127/158) and 71.4% (115/161) for devices of first and third use, respectively (P = 0.06). We concluded that the CIDR-G with short-term protocol was effective for estrous synchronization and ovulation, with lower serum progesterone concentrations for reused devices. Three times used CIDR-G yielded a pregnancy rate >70%, which tended to be lower than that obtained with new devices, adding evidence of the detrimental effect of low serum progesterone concentrations on fertility in sheep.

Alternatives to improve a prostaglandin-based protocol for timed artificial insemination in sheep.

The objective was to improve the reproductive performance of a prostaglandin (PG) F(2α)-based protocol for timed artificial insemination (TAI) in sheep (Synchrovine®: two doses of 160 µg of delprostenate 7 d apart, with TAI 42 h after second dose). Three experiments were performed: Experiment 1) two doses of a PGF(2α) analogue (delprostenate 80 or 160 µg) given 7 d apart; Experiment 2) two PGF(2α) treatment intervals (7 or 8 d apart) and two times of TAI (42 or 48 h); and Experiment 3) insemination 12 h after estrus detection or TAI with concurrent GnRH. Experiments involved 1131 ewes that received cervical insemination with fresh semen during the breeding season (32/34 °S-58 °W). Estrous behaviour, conception rate, prolificacy, and fecundity (ultrasonography 30-40 d), were assessed. In Experiment 1, ewes showing estrus between 25 and 48 h or at 72 h after the second PGF(2α) did not differ between 80 and 160 µg of delprostenate (73 vs 86%, P = 0.07; and 92 vs 95%, P = NS, respectively). Conception rate and fecundity were lower (P < 0.05) using 80 vs 160 µg (0.24 vs 0.42, and 0.27 vs 0.47, respectively). In Experiment 2, giving PGF(2α) 7 d apart resulted in higher (P < 0.05) rates of conception (0.45 and 0.51) and fecundity (0.49 and 0.53) than treatments 8 d apart (conception: 0.33 and 0.29; fecundity: 0.33 and 0.34) for TAI at 42 and 48 h, respectively. In Experiment 3, rates of conception, prolificacy and fecundity were similar (NS) between Synchrovine® with TAI at 42 h (0.50, 1.13, and 0.56) and AI 12 h after estrus detection (0.47, 1.18, and 0.55), and Synchrovine® plus GnRH at TAI (0.38, 1.28, and 0.49). However, all TAI treatments had lower (P < 0.05) prolificacy and fecundity compared to AI following detection of spontaneous estrus (1.39 and 0.83, respectively). In conclusion, the Synchrovine® protocol was: a) more successful using 160 vs 80 µg delprostenate; b) more successful with a 7 d than 8 d PGF(2α) interval; c) similarly effective for TAI versus AI 12 h after estrus detection; and d) not improved by giving GnRH at TAI.

Re-use of intravaginal progesterone devices associated with the Short-term Protocol for timed artificial insemination in goats.

Because intravaginal devices impregnated with 0.3 g of progesterone (i.e., CIDR-G) contain remaining hormone after their use in a Short-term Protocol (5 to 7 d of treatment), the reuse of these devices is proposed in goats. Two experiments were designed to study the effects of the reutilization of CIDR-G, establishing serum progesterone concentrations, follicular development, ovulatory response, and fertility. Experiment 1: Thirty dairy goats received a Short-term Protocol for 5 d using CIDR-G of first use (new devices, n = 10), second use (previously used for 5 d, n = 10), or third use (previously used twice for 5 d each time, n = 10). Goats were given (im) prostaglandin F(2α) (10 mg dinoprost) and eCG (300 IU) at device insertion and withdrawal, respectively. Serum progesterone concentrations induced by CIDR-G of first use were higher than CIDR-G of second or third use (P < 0.05); concentrations were consistently > 1 ng/mL in all females treated with reused devices. Estrus and ovulation were synchronized in 100% of goats (no differences among treatments). All females treated with new devices, but only 80% of females treated with re-used devices ovulated a new follicle that emerged after CIDR-G insertion (P = NS). Ovulation occurred between 60 and 70 h after device removal (no differences among groups). Experiment 2: In goats subjected to a Short-term Protocol followed by AI at 54 h after CIDR-G, pregnancy rates with CIDR-G of first, second, and third use were 75.3% (64/85), 67.4% (60/89), and 62.1% (54/87), respectively (devices of first versus third use, P < 0.05). In summary, intravaginal devices originally containing 0.3 g of progesterone appeared effective to synchronize estrus and ovulation after first, second and third use in the Short-term Protocol. Although the pregnancy rate with reused devices was acceptable (i.e., > 60%), it was significantly lower than that achieved with new devices and further studies to ensure adequate follicular turnover are required.

New approaches to superovulation and embryo transfer in small ruminants.

The present paper reviews the current state of knowledge in multiple ovulation and embryo transfer (MOET) technology in small ruminants, focusing on recently reported information. Major new findings are related to follicular wave patterns in small ruminants, the elucidation of follicular dominance and the integration of this information into ovarian superstimulatory treatment protocols. Follicular dynamics determine steroid and gonadotrophin secretion, follicular responses to FSH, ovulatory responses and embryo yield. Protocols that control follicular dominance have been designed to allow the initiation of superstimulation at the beginning of a follicular wave. New approaches consist of (1) synchronisation of ovulation for superstimulation during Wave 1 (i.e. Day 0 protocol), (2) pretreatment with a gonadotrophin-releasing hormone (GnRH) antagonist from 10 days prior to FSH treatment to avoid follicular dominance and (3) progesterone-oestradiol cotreatment to synchronise follicle wave emergence. These protocols provide a homogeneous pool of small follicles that are gonadotrophin responsive, enhancing the superovulatory response and embryo yield with a reduction in the incidence of unovulated follicles and early regression of corpora lutea. In addition, the rate of fertilisation failure has been reduced by using an inducer of ovulation (i.e. GnRH) associated with intrauterine insemination. In summary, the application of recently acquired knowledge has resulted in relevant improvements in MOET programmes in small ruminants.

Pregnancy rate obtained with short-term protocol for timed artificial insemination in goats.

Recent studies demonstrated that the Short-term Protocol of 5 days of progestogen treatment plus one dose of prostaglandin F(2alpha) and equine chorionic gonadotrophin (eCG) resulted in a close synchronized ovulation (60 h after the end of treatment approximately). In addition, oestradiol benzoate (ODB) is effective in synchronizing ovulation in goats and could be an alternative to eCG. This study was performed to determine the pregnancy rate using the Short-term Protocol comparing: (i) two different moments of timed artificial insemination (TAI) after eCG treatment (trial 1) and (ii) ODB as an alternative to eCG treatment (trial 2). In trial 1, 250 IU of eCG was given at the end of progestogen exposure, and cervical TAI with fresh semen was performed 48 h (n = 156) or 54 h (n = 168) later. In trial 2, 250 IU of eCG was given at sponge withdrawal (eCG group, n = 154) or 200 mug of ODB was given 24 h later (ODB group, n = 119). TAI was performed 54 h after the end of progestogen treatment. Pregnancy rate was determined by transrectal ultrasonography. In trial 1, the pregnancy rate for goats with TAI performed at 54 h (107/168, 63.7%) was higher than for those with TAI performed 48 h (77/156, 49.4%; p < 0.05) after sponge withdrawal. In trial 2, pregnancy rate was higher in eCG (94/154, 61.0%) than in ODB (49/119, 40.3%; p < 0.05) treated goats. In conclusion, the highest pregnancy rate was achieved using Short-term Protocol associated with eCG and TAI performed 54 h after treatment.

Day 0 protocol: superstimulatory treatment initiated in the absence of a large follicle improves ovarian response and embryo yield in goats.

A new superstimulatory protocol (Day 0 Protocol) to initiate FSH treatment in the absence of a large follicle was compared to a traditional protocol in goats. The Day 0 Protocol (n=44) consisted of pre-treatment with progesterone and eCG to synchronize ovulation and the emergence of Wave 1, with FSH starting 84 h after the end of progesterone exposure (i.e., soon after ovulation). The traditional protocol (n=46) consisted of 11 d of progesterone exposure, with FSH treatment beginning 2 d before the end of progesterone exposure. Treatment with FSH was initiated in the absence of a large follicle in 37/44 and in 6/46 goats in the Day 0 Protocol and traditional protocol, respectively (P<0.01). There was more CL in the Day 0 Protocol than in the traditional protocol (breeding season: 9.6+/-0.6 and 6.3+/-0.8, P<0.05; non-breeding season: 14.3+/-1.5 and 10.7+/-1.5; P<0.05). More Grades 1 and 2 embryos were recovered in the Day 0 Protocol than in the traditional protocol (breeding season: 4.8+/-0.7 and 1.8+/-0.5, P<0.05; non-breeding season: 5.6+/-1.1 and 3.5+/-0.7, P=0.07). Similarly, the proportion of embryos that were Grades 1 and 2 was higher for the Day 0 Protocol than for the traditional protocol (breeding season: 81/114, 71%, versus 16/43, 37%, P<0.05; non-breeding season: 118/203, 58% versus 95/205, 46%, P<0.05). In summary, the Day 0 Protocol, was effective in initiating superstimulatory treatment in the absence of a large follicle, and compared to the traditional protocol, induced a higher ovulation rate and better embryo yield in goats.

Endocrine, luteal and follicular responses after the use of the short-term protocol to synchronize ovulation in goats.

The effect of the so-called Short-Term Protocol (5-day progesterone treatment+PGF(2)alpha) on ovarian activity and LH surge was studied in goats. The goats received 250IU eCG at the time of device withdrawal (eCG group; n=7), or 200microg of EB (estradiol benzoate) 24h after device withdrawal (EB group; n=8), or received neither eCG nor EB (control group; n=8). The Short-Term Protocol induced greater (4.1+/-1.1ng/ml) progesterone serum concentrations at 24h after start of the treatment, that declined to 0.2+/-0.1ng/ml at 12h after device withdrawal. In all of the groups, the maximum concentration of estradiol-17beta was reached at about 36h after device withdrawal. Maximum concentration was greater in the EB group (76.9+/-24.6pmol/l) than in the control group (41.8+/-9.0pmol/l; P<0.01), with the eCG group showing intermediate concentration (70.3+/-32.5pmol/l; P=NS). The LH peak occurred earlier in the eCG group (38.4+/-2.0h after device withdrawal) and in the EB group (41.0+/-4.1h), than in the control group (46.3+/-5.1h; P<0.05). Ovulation occurred earlier in the eCG group (5/7) and in the EB group (8/8) (58.8+/-2.7h and 63.0+/-5.6h, respectively), than in the control group (7/8) (70.2+/-8.3h; P<0.05). In summary, the Short-Term Protocol induced similar concentrations of progesterone among treated goats. In addition, eCG or EB resulted in a similar increase in estradiol-17beta and a similar LH surge, which induced ovulation in most females (86.7%) in a consistent interval (about 60h) after the end of progesterone exposure.

Endocrine and ovarian changes in response to the ram effect in medroxyprogesterone acetate-primed Corriedale ewes during the breeding and nonbreeding season.

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4-6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17beta were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17beta concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17beta, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17beta levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17beta concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.

Prostaglandin F2alpha treatment associated with timed artificial insemination in ewes.

Traditional treatments of two prostaglandin F2 alpha (PGF2alpha) doses at 10-day intervals or more did not result in acceptable pregnancy rates in timed artificial insemination (TAI) programmes in ewes. An explanation might be the undefined time-period of the onset of oestrus and ovulation after the treatment. Recently a consistent interval to oestrus and ovulation was obtained by giving PGF2alpha at day 3 post-ovulation, i.e. when the largest follicle of the first follicular wave of the cycle was still growing. This can be achieved when a second dose of PGF2alpha is given 7 days after a first dose. In this work, we evaluated the synchronization of oestrus and determined which of three different moments of TAI was the most successful using a PGF2alpha (PG-7d) protocol in a large flock. A total of 436 nulliparous and multiparous ewes were treated with two doses of a PGF2alpha analogue (delprostenate 160 microg, i.m.) separated by 7 days. Onset of oestrus was recorded twice a day and a single cervical TAI with fresh undiluted semen was performed either at 42 h (n = 152), 48 h (n = 120), or 54 h (n = 164), after the second PGF2alpha dose without taking into account the oestrous response. Pregnancy rate was determined by transrectal ultrasonography 30 days after insemination. Onset of oestrus was detected in 308 of 328 and 89 of 108 multiparous and nulliparous ewes, respectively (p < 0.001), within 72 h after treatment. The distribution of the onset of oestrus did not differ between multiparous and nulliparous ewes and the highest proportion of ewes in oestrus was detected between 25 to 48 h (313/397) from the second PGF2alpha dose. The pregnancy rate in ewes inseminated at 42 h tended to be higher than those inseminated at 48 h (p = 0.09) and was higher than those inseminated at 54 h (p < 0.05) (56/152, 31/120, 37/164; respectively). Therefore, the use of the PG-7d protocol resulted in a very high synchronization of oestrus with the highest concentration (around 80%) between 25 to 48 h from the end of treatment. The best pregnancy rate (37%) was obtained after a single cervical TAI with fresh semen at 42 h.

Overview of the response of anoestrous ewes to the ram effect.

The present review summarises knowledge of the reproductive response of anoestrous ewes to the introduction of rams - in other words, the ram effect. The ovarian and endocrine response, the factors that determine whether ewes will respond or not (associated with both the stimulus and the receptivity of the ewes) and some aspects of practical management are discussed. Information on the use of the ram effect to stimulate post-partum, prepubertal and cyclic ewes is also given. New insights are provided on ovarian response patterns, including recently collected information on luteal responses. The existence of delayed ovulation (5-7 days after the introduction of the rams) followed by luteal phases of normal or short length, luteal cysts and luteinised follicles is reported after scanning the ovaries with ultrasound. Endocrine parameters for depth of anoestrus, such as LH pulsatility and FSH concentrations, and how the concentrations of these hormones should be considered are discussed. Particular attention is paid to the observation of spontaneous, higher LH pulsatility and higher FSH concentrations in anoestrous ewes that respond to rams with luteal phases than in those that fail to respond. The use of progestogen priming and single progestogen administration and the possible advantages for synchronisation of oestrus are also discussed. Other factors that should be considered before the ram effect is applied, such as the strength of the stimulus and some practical considerations, are also included.

Response of the 1-5 day-aged ovine corpus luteum to prostaglandin F2alpha.

The hypothesis that, in the ewe, prostaglandin (PG) F2alpha administration on day 3 after ovulation is followed by luteolysis and ovulation was tested using 24 animals. The ewes were treated with a dose of a PGF2alpha analogue (delprostenate, 160 microg) on days 1 (n=8), 3 (n=8) or 5 (n=8) after ovulation, was established by transrectal ultrasonography. Daily scanning and blood sampling were performed to determine ovarian changes and progesterone serum concentrations by radioinmunoassay. The treatment induced a sharp decrease of progesterone concentrations followed by oestrus and ovulation in all ewes treated on days 3 and 5 and in one ewe treated on day 1 (8/8, 8/8, 1/8; P<0.05). Seven ewes treated on day 1 did not respond to PGF2alpha treatment and had an inter-ovulatory cycle of normal length (17.4 +/- 0.5 days). However, the profile of progesterone concentrations during the cycle of these ewes was delayed 1 day (P<0.05) compared with a control cycle. The overall interval between PGF2alpha and oestrus for the 17 responding ewes was 42.4 +/- 2.3 h. In 15 of these ewes the ovulatory follicle was originated from the first follicular wave and the ovulation occurred at 60.8 +/- 1.8 h after PGF2alpha treatment. The other two responding ewes ovulated an ovulatory follicle originated from the second follicular wave between 72 and 96 h after treatment. These results support the hypothesis and suggest that refractoriness to PGF2alpha of the recently formed corpus luteum (CL) may be restricted to the first 1-2 days post-ovulation.

Effect of high progesterone concentrations during the early luteal phase on the length of the ovulatory cycle of goats.

The effect of exogenous progesterone exposure early in the oestrous cycle on the duration of the interovulatory interval was studied in dairy goats. A controlled intravaginal drug release (CIDR-G) device was inserted for 5 days starting at day 0 (D0 group, n=6) or day 3 (D3 group, n=5) postovulation. A third group was composed of untreated control goats (control group, n=7). Daily transrectal ultrasound was carried out during the interovulatory interval to assess the ovarian dynamics. Oestrous behaviour was checked twice a day and serum progesterone levels were assayed in daily jugular blood samples. Treated goats showed two different responses. In three D0 goats and one D3 goat, progesterone concentrations fell immediately after CIDR withdrawal and this was followed by oestrus and ovulation between days 8 and 11 (short cycles). In the other three D0 goats and in four D3 goats the treatment significantly reduced the interovulatory interval (18.3+/-0.3 and 18.5+/-0.3 days, respectively) (shortened cycles) compared with the control group (20.0+/-0.2 days; P<0.05), but the intervals with progesterone concentrations over 1 ng/ml were not different (15.7+/-0.3, 15.8+/-0.7 and 16.0+/-0.5 days for D0, D3 and control goats, respectively). In all D0 goats with a short cycle response, the ovulatory follicle arose from the first follicular wave but in the D3 goat with a short cycle it arose from the second follicular wave. These results showed that premature progesterone exposure early in the ovulatory cycle of the goat affected its length inducing short or shortened cycles. The effect of progesterone could either affect luteotropic support of the corpus luteum (CL) and/or stimulate a premature release of the luteolysin.

Circulating gonadotrophins and follicular dynamics in anestrous ewes after treatment with estradiol-17beta.

Plasma FSH, LH, estradiol (E2) and progesterone (P4) profiles and patterns of follicular growth and regression by ultrasonography were determined after E2 treatment (1 microg/kg) in anestrous ewes. Fifteen ewes were treated with one (group I, n=7) or two (group II, n=4) i.m. injections of E2 with a 24h interval, or two oil injections with a 24h interval (group C, n=4). Blood samples for E2, P4, FSH and LH determinations were collected daily 4 days before the initiation of the treatment (day 0), when bleeding increased to every 2h starting 2h before treatment until 56h after the first injection and from then on every 6h until day 8, and twice per day till the end of the experiment (day 9). During the experimental period (days -4 to 9), transrectal ultrasonic examinations were carried out daily using a 7.5 MHz linear array probe. Number and size of follicles > or =3mm in diameter were recorded. No estrous was detected before, during or after treatment. LH and FSH surges were observed 10-18h after the first E2 injection. The second E2 injection stimulated another release of LH but no surges. E2 inhibited FSH levels before the surge and the second E2 injection induced a longer inhibition. No ovulation was detected by ultrasonography during the experimental period and P4 levels remained low (<0.7 nmol/l) before, during and after the treatment in all ewes. There was an effect of E2 treatment on the diameter of the largest follicle, a decrease could be observed 3 days after the first injection in both ewes of groups I and II. The E2-treated groups had a higher frequency of ewes showing wave emergence on day 3 (day 1.5+/-1,2.4+/-0.4 and 2.5+/-0.5 for control, groups I and II). LH and FSH surges were observed after E2 treatment, but were not able to provoke ovulation neither luteinization. In contrast, the treatment was associated with the regression of the largest follicle and with emergence of a new follicular wave on day 3.

Effect of long-term and short-term progestagen treatment on follicular development and pregnancy rate in cyclic ewes.

The aim of this study was to evaluate the effect of the length of a progestagen treatment (12 d vs. 6 d) on follicular dynamics, estrus synchronization and pregnancy rate using medroxyprogesterone acetate (MAP) with or without an eCG dose at the end of MAP treatment. One hundred sixty Polwarth ewes were divided into four equal groups: long-term treated (LT, n=40); short-term treated (ST, n=40); long-term treated plus eCG (LTeCG, n=40); and short-term treated plus eCG (STeCG, n=40). Five ewes of each group were separated to undergo daily transrectal ultrasonography, and blood samples were taken for hormone determination. Until 96 h after sponge withdrawal the number of ewes in estrus was higher in both long-term-treated groups than in both short-term-treated groups but at the end of the observational period (144 h) no significant differences were found among groups. The pregnancy rate was higher in the ST group (87%) than in the other groups (LT, 63%; LTeCG, 67%; and STeCG, 58%; P< or =0.03). The ovulatory follicle emerged before sponge withdrawal in long-term-treated ewes (-3.8+/-0.4 d and -2.2+/-0.8 d for LT and LTeCG, respectively), whereas in short-term-treated ewes it emerges around sponge removal (0.4+/-1.1 d and 0.5+/-0.5 d for ST and STeCG, respectively; P< or =0.01). The ovulatory follicle in the LT group had a longer lifespan and attained a larger (P< or =0.05) maximum diameter than in the ST group. We conclude: a) that the lower pregnancy rate observed after long-term progestagen treatment was related to a slower follicular turnover that promoted the ovulation of persistent dominant follicles; (b) that short-term treatment resulted in a higher pregnancy rate probably due to the ovulation of newly recruited growing follicles; and (c) treatment with eCG had no advantage in association with long-term treatment and had a deleterious effect in combination with short-term treatment with MAP.

The effect of subluteal levels of exogenous progesterone on follicular dynamics and endocrine patterns during early luteal phase of the ewe.

Nineteen Corriedale ewes were treated with an im dose of a PGF2alpha during the luteal phase to synchronize estrus. After ovulation had been detected by using ultrasonography (Day 0); the ewes were randomly assigned to 2 different groups. In 11 ewes a CIDR, which had previously been used for 10 d, was inserted on the fourth day after ovulation. The ewes then received a dose of PGF2alpha on Day 5 to induce luteolysis. The CIDR remained in place until the end of the experiment (Day 9). Control ewes (n = 8) received no treatment. Blood samples were taken daily for estradiol, progesterone and FSH determinations. In the untreated ewes, 2 follicular waves were detected in all of the animals throughout the monitoring period, with a mean wave interval of 4.5 d. The total number of follicles which were > or =2 mm decreased from Day 0 to Day 4 (8.8+/-1.0 to 5.3+/-0.6; P< or =0.05) and then increased at Day 7 (7.5+/-0.9; P< or =0.05). The growth profiles of both the largest and the second largest follicles of Wave 1 showed significant divergence, while no divergence was observed in Wave 2. Serum estradiol concentrations decreased significantly from the day before to the day of ovulation and then increased again during the growing phase of the largest follicle of Wave 1. Concentrations of FSH were high on the day of emergence of both waves, but while a significant decline was observed after emergence in Wave 1, the levels remained high in Wave 2. In 8 of the 11 treated ewes, the largest follicle of Wave 1 was still present on the ninth day after ovulation (persistent follicle). In the other 3 ewes, the largest follicle of Wave 1 was already regressing on the day that the treatment was administered, and the largest follicle that was present on Day 9 originated from Wave 2 (nonpersistent follicle). In persistent follicle ewes, the largest follicle of Wave 1 prolonged its lifespan significantly, attaining the maximum diameter (Day 8.1+/-0.8) later than in untreated (Day 3.0+/-0.4) and nonpersisted follicle ewes (Day 2.0+/-0.6). The total number of follicles decreased in persistent follicle ewes between Day 0 and Day 4 (7.9+/-1.5 to 4.5+/-0.5, respectively; P< or =0.05) and remained low until the end of the experiment. Progesterone concentrations (nmol/L) between Days 6 and 9 were significantly different between untreated and persistent follicle ewes (12.8+/-1.0 vs. 9.4+/-1.0, P< or =0.02). The present study confirms that the largest follicle of Wave 1 is dominant in the ewe and that subluteal progesterone concentrations can prolong its lifespan and extend this dominance.

Ovarian dynamics, serum estradiol and progesterone concentrations during the interovulatory interval in goats.

Ovarian changes determined by daily transrectal ultrasonic scanning, and its correlation with serum progesterone (P4) and estradiol (E2) concentrations were studied in seven cyclic Saanen goats. Estrous cycles were synchronized with 2 injections of a PGF2 alpha analogue 9 d apart. All follicles > or = 2 mm in diameter and CL were measured each day. One goat showed a longer interestrous interval, associated with development of a cystic-luteinized structure. The mean interovulatory interval for the other 6 goats was 20.8 +/- 0.4 d. The incidence of goats with 4, 3, and 2 follicular waves was 3, 1 and 2 respectively; follicular waves emerged on Days 0.5 +/- 0.6, 7.2 +/- 0.7, 10.7 +/- 0.5 and 13.7 +/- 0.8 for Wave 1, 2, 3 and the Ovulatory wave, respectively. The largest follicle of Wave 2 was smaller (4.9 +/- 0.1 mm) than the largest follicles of Wave 3 (6.2 +/- 0.1 mm; P < or = 0.01) and of the Ovulatory wave (7.0 +/- 0.5 mm; P < or = 0.01), and tended to be smaller than the largest follicle of Wave 1 (6.3 +/- 0.6 mm; P < or = 0.09). Interval between emergence of Wave 1 and Wave 2 was longer than interval between emergence of Wave 2 and Wave 3 (7.3 +/- 0.9 d vs 4.0 +/- 0.4 d; P < or = 0.01), and between Wave 3 and the Ovulatory wave (3.8 +/- 1.1 d; P < or = 0.05). Two days before ovulation, the diameter of the ovulatory follicle was larger (P < or = 0.01) than the first subordinate follicle. Serum E2 concentrations increased from the day of ovulation (2.7 +/- 0.3 pg/mL) to Day 2 (7.6 +/- 0.9 pg/mL; P < or = 0.01), associated with the early-mid growing phase of the largest follicle of Wave 1, and then decreased to basal levels on Day 5 (P < or = 0.01) and peaked again (16.5 +/- 2.4 pg/mL) 2 d before ovulation. The CL were detected ultrasonically on Day 3 post ovulation and attained a mean maximum diameter of 13.5 +/- 0.8 mm between Days 8 and 14. The following characteristics were observed: 1) ovarian follicular development in goats is wave-like; 2) increased P4 concentrations may be promoting follicular wave turnover; 3) it is suggested that the presence of follicular dominance and the production of E2 are different among waves. While in Wave 1 and in the Ovulatory wave, follicular dominance is present and production of E2 is consistent, no changes in serum E2 concentrations were found in other stages of the interovulatory interval. In the intervening waves, no indicators of follicular dominance could be firmly documented.

Response of Corriedale ewes to the "ram effect" after priming with medroxyprogesterone, fluorogestone, or progesterone in the non-breeding season.

One hundred eighty-nine Corriedale ewes were used during the non-breeding season to study the "ram effect" stimulus after priming with progestogens. Intravaginal sponges containing either medroxyprogesterone acetate (MAP group, n = 49), fluorogestone acetate (FGA group, n = 49), or progesterone devices (CIDR group, n = 46) were inserted on Day-6 (Day 0 = introduction of the rams). Forty-five ewes were untreated and kept as a control group. On Day 0 the sponges were removed and rams provided with marking harnesses for oestrous detection were placed with the ewes. Onset of estrus was monitored until Day 25, and conception was determined by transrectal ultrasonography. Ewes came into heat during 4 periods: Days 0-3, 5-7, 17-20, and 21-23. The overall number of oestrus ewes were 29%, 53%, 35%, and 50% for the control, MAP, FGA, and CIDR groups, respectively (MAP and CIDR > control, p < 0.05). Control ewes presented oestrus only on Days 17-20 and 21-23. Oestrus in the progestogen-primed ewes was concentrated during Days 0-3 and 17-20, and some ewes came into oestrus on Days 5-7. There were no differences between different primings neither in oestrous response nor in conception rate. The conception rate from matings occurring on Days 0-3 was higher than on those occurring on Days 17-20. We conclude that MAP, FGA, and CIDR is equally effective in improving the response to the ram effect, and the pattern of oestrus in primed ewes was different than previously reported.

Ovarian response to gonadotropin treatment initiated relative to wave emergence in ultrasonographically monitored ewes.

Follicular recruitment and luteal response to superovulatory treatment initiated relative to the status of the first wave of the ovine estrous cycle (Wave 1) were studied. All ewes (n = 25) received an intravaginal progestagen sponge to synchronize estrous cycles, and ewes were monitored daily by transrectal ultrasonography. Multiple-dose FSH treatment (total dose = 100 mg NIH-FSH-P1) was initiated on the day of ovulation (Day 0 group) in 16 ewes. In the remaining 9 ewes, FSH treatment was started 3 d after emergence of the largest follicle of Wave 1 (Day 3 group). Ewes received PGF(2alpha) with the last 2 FSH treatments to induce luteolysis. Daily blood samples were taken to determine progesterone profiles and to evaluate the luteal response subsequent to superovulation. The ovulation rate was determined by ultrasonography and correlated with direct observation of the ovaries during laparotomy 5 to 6 d after superovulatory estrus when the uterus was flushed to collect embryos. Results confirmed that follicular recruitment was suppressed by the presence of a large, growing follicle. In the Day 0 and Day 3 groups, respectively, mean numbers (+/- SEM) of large follicles (>/= 4 mm) recruited were 6.4 +/- 0.6 and 2.7 +/- 0.7 (P < 0.01) at 48 h after the onset of treatment, and 6.7 +/- 0.5 and 5.1 +/- 0.6 (P = 0.08) at 72 h after the onset of treatment. Ovulation rates were 5.6 +/- 0.8 and 3.3 +/- 0.8 in the respective groups (P < 0.05). The number of transferable embryos was 1.8 +/- 0.5 and 0.3 +/- 0.2 in the respective groups (P < 0.05). Short luteal phases (