No adverse effect of confirmation of ovulation by rectal palpation and ultrasonography after artificial insemination on formation, development, and function of the corpus luteum and conception rate in cows.

The effect of confirmation of ovulation by rectal palpation and ultrasonography after artificial insemination (AI) on the development of the corpus luteum (CL) and conception rate was investigated in cows. A total of 90 clinically healthy Holstein-Friesian dairy cows were examined in this study. After AI, the cows were divided into three groups (30 cows per group). In Group I, ovulation was confirmed by rectal palpation at 24 h after AI. In Group II, ovulation was confirmed using transrectal ultrasonography 24 h after AI. In Group III, ovulation was not confirmed after AI. Day 0 was defined as the day when ovulation was confirmed in Groups I and II, and as the day after AI was performed in group III. Transrectal ultrasonography was performed on days 3, 5, 7, and 14 to measure the CL diameter, tissue area, and CL blood flow area, and the ratio of CL blood flow area to CL tissue area was calculated. On the day of CL measurement, blood samples were collected to determine the plasma concentrations of progesterone (P4) and estradiol-17ß (E2). Pregnancy was diagnosed at 28 and 60 days after AI. A high conception rate of approximately 80% was achieved in Groups I and II, in which confirmation of ovulation was conducted. There were no differences in the diameter, tissue area, or blood flow area of the CL between the three groups. These results indicate that the confirmation of ovulation by rectal palpation and transrectal ultrasonography did not affect the formation and function of the CL or conception rate.

Changes in the ovary and the peripheral concentrations of sex hormones after the aspiration of follicular fluid from the spontaneous follicular cysts of dairy cows.

The aim of the present study was to clarify the ovarian and hormonal dynamics after the aspiration of follicular fluid in cows with follicular cysts. Follicular fluid was aspirated from the follicular cysts and follicles that were fated to become cystic follicles and other coexisting normal follicles, respectively, in lactating cows (n = 3). After the aspiration procedure, new follicles developed and reached a diameter of 25 mm without ovulation within 13-19 days. The plasma concentrations of inhibin decreased and follicle-stimulating hormone increased rapidly after the aspiration procedure, and subsequently increased and decreased, respectively, as a new follicle grew. No luteal structures developed after the aspiration procedure, and the animals' plasma progesterone levels remained low. The present study indicates that the cystic follicles are never luteinized by the aspiration of follicular fluid, and consequently, new follicular cysts are observed to repeatedly develop.

An investigation of the time period within which frozen-thawed semen delivers a high conception rate in lactating dairy cows.

We determined the length of time within which frozen-thawed semen delivered a high conception rate in present-day lactating dairy cows. The cows utilized were a total 100 milking Holstein-Friesian cows kept in tie-stall style farms. We carried out artificial insemination (AI) during the periovulatory period at a predicted time based on ovulation, and checked ovulation at 6-h intervals after AI. The period from AI to ovulation ranged from 48 h (i.e., 48 h before ovulation) to -12 h (i.e., 12 h after ovulation). High conception rates averaging 63.0% were obtained by carrying out AI > 6-30 h before ovulation, significantly higher than the conception rates of 30.0% (P < 0.05) and 26.9% (P < 0.01) from AI carried out earlier than 30 h before ovulation and later than 6 h before ovulation, respectively. It was concluded that frozen-thawed semen delivers a conception rate of ≥ 60% for > 24-30 h after AI, and that a conception rate of ≥ 60% can be achieved by carrying out AI 6-30 h before ovulation using frozen-thawed semen.

Evaluation of criteria for optimal time AI postulated by estrous signs in lactating dairy cows kept in tie-stalls.

Relaxation of the intravaginal part of the uterus is obvious around 6 to 18 h before ovulation, and this is considered the optimal time for artificial insemination (AI), as demonstrated in recent studies. Estrous signs have been suggested as useful criteria for determining the optimal time for AI. Therefore, this study evaluated the usefulness of estrous signs, particularly the relaxation of the intravaginal part of the uterus, as criteria for determining the optimal time for AI. A Total of 100 lactating Holstein-Friesian cows kept in tie-stall barns were investigated. AI was carried out based on the criterion for the optimal time for AI (optimal group), and earlier (early group) and later (late group) than the optimal time for AI, determined on the basis of estrous signs. After AI, ovulation was assessed by rectal palpation and ultrasonographic observation at 6-h intervals. For 87.5% (35/40) of cows in the optimal group, AI was carried out 24-6 h before ovulation, which was previously accepted as the optimal time for AI. AI was carried out earlier (early group) and later (late group) than optimal time for AI in 62.1% (18/29) and 71.0% (22/31) of cows, respectively. The conception rate for the optimal group was 60.0%, and this conception rate was higher than that for the early group (44.8%) and late group (32.2%), without significance. Further, the conception rate of the optimal group was significantly higher than the sum of the conception rates of the early and late groups (38.3%; 23/60) (P < 0.05). These results indicate that the criteria postulated, relaxation of the intravaginal part of the uterus and other estrous signs are useful in determining the optimal time for AI. Furthermore, these estrous signs enable the estimations of stages in the periovulatory period.

Relationships between the appearances and changes of estrous signs and the estradiol-17ß peak, luteinizing hormone surge and ovulation during the periovulatory period in lactating dairy cows kept in tie-stalls.

Lactating Holstein-Friesian cows kept in tie-stall barn were used as subjects in this study. Rectal examination, ultrasonography and blood sampling were conducted every other day and then daily after the day on which diameter of the corpus luteum decreased. After the luteal diameter decreased for 2 consecutive days, rectal and ultrasound examinations, blood sampling, and observation of estrous signs were conducted at 6-h intervals. Most of the estrous signs became obvious with the increase in estradiol-17ß (E2) and became most remarkable 24 to 30 hours before ovulation, at which point the E2 peak and luteinizing hormone (LH) surge were achieved, and then weakened which progression to ovulation. The correlation between the intensity of four estrous signs (hyperemia and swelling of the intravaginal part of the uterus, opening of the external uterine orifice and viscosity of the cervical mucus) and the plasma E2 concentration was higher than that of three estrous signs (swelling of the vulva, contraction of the uterus, diameter of uterine horn) and the plasma E2 concentration. The relaxation of the intravaginal part of the uterus showed a unique change compared with the other estrous signs, and it became most obvious 6, 12 and 18 h before ovulation; this obviously relaxed period was consistent with the generally accepted theoretical optimal time for artificial insemination (AI), i.e., 6 to 24 h after initiation of estrus. These results suggest that observation of estrous signs by vaginoscopic examination gave useful information for detection of the optimal timing of AI in the periovulatory period in lactating dairy cows kept in a tie-stall barn.

Progesterone profiles in the caudal vena cava and jugular vein in response to pulsatile luteinizing hormone stimulation induced by GnRH treatment during the mid-luteal phase in lactating dairy cows.

The aim of this study was to examine whether increased frequency of luteinizing hormone (LH) pulses influences luteal progesterone (P4) secretion by measuring progesterone concentrations at the secreted (caudal vena cava) and circulating levels (jugular vein) in lactating dairy cows. Cows received six intravenous administrations of 2.5 µg of GnRH (gonadorelin acetate, n=4) or 2 ml saline (n=3) at 1-h intervals on 12.4 ± 0.4 (mean ± SE) days after ovulation. Blood samples were collected from the caudal vena cava and jugular vein every 12 min for 12 h (6 h before and after treatment). During the 6 h after treatment, frequency of LH pulses (5.3 ± 0.3 and 3.0 ± 0.0 pulses/6 h) and mean LH concentration (0.50 ± 0.06 and 0.38 ± 0.05 ng/ml) were greater (P<0.05) in GnRH-treated cows than in saline-treated cows. Mean P4 concentration and amplitude of P4 pulses in the caudal vena cava during the 6 h after treatment were greater (P<0.05) in GnRH-treated cows than in saline-treated cows, but the frequency of P4 pulses was not different between the groups. Mean P4 concentration in the jugular vein during the 6 h after treatment was also higher (P<0.05) in GnRH-treated cows than in saline-treated cows (7.0 ± 1.3 and 5.4 ± 0.9 ng/ml). These results indicate that the increased frequency of LH pulses stimulates progesterone secretion from the functional corpus luteum and brings about higher P4 concentrations in the circulating blood in lactating dairy cows.

Changes in plasma progesterone levels in the caudal vena cava and the jugular vein and luteinizing hormone secretion pattern after feeding in lactating and non-lactating dairy cows.

The present study was designed to assess progesterone profiles at the secreted (caudal vena cava) and circulating levels (jugular vein) and luteinizing hormone (LH) secretion pattern in lactating and non-lactating cows with reference to feeding. Four lactating and four non-lactating cycling Holstein cows were examined. Blood samples were collected simultaneously from the caudal vena cava (via a catheter inserted from the coccygeal vein) and the jugular vein every 15 min for 12 h (0500-1700 h) during the functional luteal phase. Cows were fed 50% of the daily diet 6 h after the start of blood sampling. During the 12-h sampling period, mean progesterone concentrations in the caudal vena cava did not differ between lactating and non-lactating cows (49.0 ± 2.9 and 53.3 ± 3.7 ng/ml; mean ± SE), whereas mean progesterone concentrations in the jugular vein in lactating cows were higher than those in non-lactating cows (6.4 ± 0.1 and 5.6 ± 0.1 ng/ml, P < 0.001). Lactating cows had a higher frequency of LH pulses than non-lactating cows (7.0 ± 0.7 and 4.3 ± 0.9 pulses/12 h, P<0.05). The influence of feeding was not observed on LH profiles but was observed on progesterone profiles in both veins. Progesterone concentrations in the caudal vena cava increased after feeding in both groups. Progesterone concentrations in the jugular vein decreased after feeding in lactating cows but not in non-lactating cows. These results indicate the difference in feeding-related changes in progesterone dynamics between lactating and non-lactating cows.

Exposure to estrogen mimicking the level of late pregnancy suppresses estrus subsequently induced by estrogen at the level of the follicular phase in ovariectomized shiba goats.

A high-estrogen environment during late pregnancy is suspected to cause postpartum silent ovulation, and progesterone (P4) is suggested to recover estrus. However, few attempts have been undertaken to elucidate the influence of these steroids on estrus by analyzing hormonal profiles. We investigated estrus and luteinizing hormone (LH) surges in ovariectomized goats (n=6) assigned to three treatments in a cross-over design. In groups 1 and 2, 200 µg/kg body weight/day estradiol benzoate (Dose-200 E2B) was administered for 14 days concurrent with P4 for 11 days, while in the control, saline solution and P4 were administered likewise. Ten days after the final administration of Dose-200 E2B, group 2 was treated with P4 for 8 days, and all groups were treated with 2 µg/kg body weight E2B (Dose-2 E2B) 20 days after the final administration of Dose-200 E2B (or saline solution). The proportion of cases expressing estrus after the administration of Dose-2 E2B was smaller (P<0.01) in group 1 than in the control (1/6, 3/6 and 6/6; groups 1 and 2 and the control, respectively). The proportions of cases generating LH surges did not differ (P>0.1) among the groups (5/6, 5/6 and 6/6; groups 1 and 2 and the control, respectively), but the peak concentrations in groups 1 and 2 (26.2 ± 14.7 and 11.3 ± 6.7 ng/ml) were lower (P<0.01) than those in the control (67.8 ± 19.4 ng/ml). These results demonstrated that elevation of plasma estrogen mimicking late pregnancy inhibits the subsequent estrus induced by estrogen simulating the follicular phase.

Comparison between lactating and non-lactating dairy cows on follicular growth and corpus luteum development, and endocrine patterns of ovarian steroids and luteinizing hormone in the estrous cycles.

The dynamics of ovarian follicle, corpus luteum (CL), and peripheral plasma ovarian steroids were compared between lactating and non-lactating cows, and a possible association of pulsatile luteinizing hormone (LH) secretion with the dynamics was examined. Lactating (n=5) and non-lactating (n=5) cows were monitored daily for follicle and CL throughout two consecutive estrous cycles (Day 0: day of ovulation). Blood samples were collected daily and at 15 min intervals for 8h on Days 2, 4, 6, 8, and 14 of the second cycle. Lactating cows had larger CL (25.4 ± 1.8mm vs. 23.5 ± 1.5mm, P<0.01) and greater progesterone concentrations (4.6 ± 1.0ng/ml vs. 3.9 ± 0.9 ng/ml, P<0.01) during mid-luteal phase compared with non-lactating cows. Maximal diameters of the first wave dominant follicle (17.2 ± 1.8mm vs. 15.5 ± 0.8mm) and the ovulatory follicle (17.9 ± 1.2mm vs. 15.2 ± 0.8mm) were greater (P<0.05) in lactating cows than in non-lactating cows during the estrous cycles with two follicular waves, but no significant differences were detected between the groups during the estrous cycles with three follicular waves. Plasma estradiol concentrations did not differ between the groups throughout the experiment. Lactating cows had more LH pulses from Days 2 to 14 than non-lactating cows. These results imply that differences in ovarian dynamics may exist between lactating and non-lactating cows, for which the increased number of LH pulses observed in lactating cows may have responsibility.

Profiles of ovarian steroids, luteinizing hormone and estrous signs from luteolysis to ovulation in lactating and non-lactating dairy cows.

The aims of the present study were to investigate the profiles of ovarian steroids and luteinizing hormone (LH) and the appearance of estrous signs in relation to luteolysis and ovulation in lactating and non-lactating cows and to examine the influence of lactation on those observations. Five lactating (daily milk yield of 28.4 ± 3.2 kg; mean ± SD) and five non-lactating cycling Holstein cows were examined. Their ovaries were monitored by ultrasonography daily during one estrous cycle. Blood samples were collected daily and then at 3-h intervals after luteolysis until ovulation. Estrous signs in terms of behavior, the vulva and the vagina were checked at 8-h intervals after luteolysis until ovulation. Profiles of progesterone, estradiol-17ß and LH did not differ between the groups. There were no differences in the interval from luteolysis to ovulation (4.6 ± 0.5 and 4.2 ± 0.8 days) and the interval from the estradiol-17ß peak to ovulation (34.2 ± 4.5 and 30.6 ± 3.9 h) between lactating and non-lactating cows. The interval from the peak of the LH surge to ovulation was 27 h in all cows examined. Appearance of estrous signs did not differ between the groups. The vaginal estrous signs were observed conspicuously in all cows examined, but the behavioral signs were not observed in 20.0% of the cows. The duration of behavioral signs (41.3 ± 23.6 h) was shorter (P<0.05) than that of the vagina (68.9 ± 25.4 h). These results imply that lactation might not interfere with the hormonal profiles from luteolysis to ovulation.

Intermittent nutritional stimulus by short-term treatment of high-energy diet promotes ovarian performance together with increases in blood levels of glucose and insulin in cycling goats.

The aim of this study was to determine if short-term intermittent treatments of high-energy diet have any stimulatory effects on ovarian function and metabolic status in goat. Cycling Shiba goats were divided into treatment (TG; n=6) and control (CG; n=6) groups. After the detection of ovulation (1st ovulation, Day 0) by ultrasonography, a high-energy diet (250% of maintenance) was fed to the TG from Day 12 to Day 15 (4 days) and from Day 18 to Day 21 (4 days). The high-energy diet comprised 1000 g hay-cubes and 300 g of concentrated feed/head/day (approximately 15 MJ of digestible energy/day). The CG was offered maintenance diet throughout the experiment. Transrectal ultrasound examinations were conducted every other day during the luteal phase and were conducted daily during the follicular phase. Blood samples were collected daily from Day -2 to 7 days after ovulation (2nd ovulation) following the nutritional treatment for analysis of follicle stimulating hormone (FSH), progesterone, oestradiol, glucose and insulin in plasma. Two wave-like rises in the concentrations of glucose and insulin appeared in response to the intermittent nutritional stimulus. Mean plasma concentrations of glucose and insulin were significantly (P<0.05) higher in the TG than in the CG on almost all days during each treatment period. For ovarian performance, the total number of follicles that developed on the day prior to the 2nd ovulation in the TG (7.5 ± 2.7) was significantly higher than that of the CG (4.5 ± 2.6). Ovulation rate of the 2nd ovulation (4.5 ± 1.0) in the TG was significantly higher than that of the 1st ovulation (3.0 ± 0.6) of the TG and than both 1st (2.5 ± 0.5) and 2nd (2.7 ± 0.5) ovulations of the CG. No significant difference was detected in the concentrations of FSH, progesterone and oestradiol throughout the experiment between TG and CG. The present experiment demonstrated that short-term intermittent nutritional stimulus from the luteal phase increased the total number of ovulatory follicles and the ovulation rate in association with increasing plasma concentrations of glucose and insulin in goats.

Responses of gonadotropin secretion to short-term dietary supplementation in ovariectomized goats with different body weights.

The responses of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion to acute dietary supplementation were studied in goats with different body weights. Ovariectomized Shiba goats (n=11) were used and were maintained with a feed of 100% of their energy requirement. They were implanted subcutaneously with an oestradiol capsule and were divided into light (LBW; 24kg, n=5, mean+/-S.D., 32.0+/-6.3kg) groups on the basis of their body weights at 8 days before the start of treatment. At the start of treatment (Day 1), the level of a feed changed to 250% of their energy requirement and this level was maintained for 7 days in both groups. Blood samples were collected daily from Day -7 to Day 7 for the analysis of FSH, glucose, and insulin profiles in plasma. Frequent blood samples were also collected at 10min intervals for 6h on Day 0, Day 3, and Day 7 for analysis of LH pulses. LH pulse frequency increased significantly on Day 3 as compared with that on Day 0 in both the HBW (7.4+/-0.5pulses/6h vs. 6.2+/-0.8pulses/6h, p<0.05) and LBW (6.5+/-0.8pulses/6h vs. 5.5+/-0.5pulses/6h, p<0.05) groups, whereas it decreased on Day 7 (HBW, 6.4+/-0.9pulses/6h; LBW, 6.3+/-1.6pulses/6h, p>0.05 vs. Day 0). Plasma glucose and insulin concentrations increased temporarily from Day 2 to Day 4 and then decreased to the level before the start of dietary supplementation in both groups. There was no significant difference in the LH pulse frequency or daily concentrations of FSH, glucose, or insulin between the HBW and LBW groups throughout the experimental period. The present study indicated that acute dietary supplementation stimulates pulsatile LH secretion in parallel with a rise of blood glucose and insulin levels. However, the influence of body weight on these responses between light and heavy animals was not observed.

Ultrasonographic uterine changes and vaginal discharges following intrauterine infusion of liquid paraffin in cows.

This study was conducted to examine uterine changes and uterocervical discharges following intrauterine infusion with liquid paraffin (LP) during the luteal phase by ultrasonic and vaginoscopic examinations in cows. Multiparous dairy cows (n=10) were infused with 50 ml physiological saline (PS group; n=5) or liquid paraffin (LP group; n=5) on day 10 or 11 after ovulation (day 0: ovulation). Vaginoscopic, rectal and ultrasonogaphic examinations were carried out at 0.25, 1, 2, 3, 4, 6, 8, 12 and 24 h after the LP and PS infusion and then at daily intervals until subsequent ovulation after the infusions. The mean volumes of recovered discharges from the vagina within 6 h after infusion were significantly greater (P<0.05) in the LP group than in the PS group (33.0 +/- 9.9 vs.14.0 +/- 13.9 ml). Yellowish-white discharge was first observed at 3.2 +/- 0.5 and 3.6 +/- 0.6 h after infusion and lasted for 12.2 +/- 2.9 and 2.1 +/- 1.5 days for the LP and PS groups, respectively, showing a significant difference (P<0.05) in duration. Subsequently, transparent discharge appeared again 2-3 days before the subsequent ovulation after the treatments in both groups and disappeared on the day prior to or the day of ovulation. During the immediate examination after the infusion, the cavity of the uterine horn appeared anechoic and dilated in the images of both groups. The anechoic images changed to echoic images at 2.2 +/- 0.8 and 2.6 +/- 0.9 h after the infusion in the LP and PS groups, respectively, and the echoic images lasted for 12.2 +/- 2.9 and 2.1 +/- 1.5 days in the LP and PS groups, respectively. These results suggest that the appearance and disappearance of intrauterine anechoic and echoic images reflect the appearance and disappearance of the characteristics of the recovered LP/PS-like liquid and yellowish-white and transparent discharges from the vagina.

The effects of short-term nutritional stimulus before and after the luteolysis on metabolic status, reproductive hormones and ovarian activity in goats.

The effect of short-term nutritional supplementation on hormonal and ovarian dynamics was studied in goats. Cycling Shiba goats were divided randomly into maintenance (group M, n=4) and high-energy (group H, n=4) groups. After the detection of the ovulation (Day 0, 1(st) ovulation), group H received a high-energy diet providing 2.5 times of the maintenance energy requirement for 7 days from Day 7 to Day 13 and were administered 2 mg of prostaglandin F(2alpha) (PGF(2alpha)) on Day 10 to induce luteal regression followed by the follicular phase. Follicular and luteal dynamics were monitored using ultrasonography daily or every other day, and blood samples were collected daily from Day 0 to the third ovulation (3(rd) ovulation) following the second ovulation (2(nd) ovulation) induced by PGF(2alpha) administration. Blood samples were also collected at 10-min intervals for 6 h on Day 9 and Day 11 for analysis of pulsatile LH secretion. The mean concentrations of glucose and insulin were significantly (P<0.05) higher in group H than in group M on Days 8, 9, 12, 13 and Days 8, 9 and 10, respectively. For both the 2(nd) and 3(rd) ovulations, no significant difference was detected in ovulation rate between groups M and H. On the other hand, the interpeak interval for wave-like patterns of FSH in group H was significantly (P<0.05) shorter than in group M during the period between the 1(st) and 2(nd) ovulations (4.3 +/- 0.3 vs. 6.5 +/- 1.5 days). The mean LH pulse frequency in group H was significantly (P<0.05) greater than in group M on Day 11 (4.5 +/- 0.6 vs. 3.3 +/- 0.5 pulses/6 h). The present study clearly demonstrated that short-term (7 days) nutritional supplementation promoted pulsatile LH and wave-like FSH secretions in cycling goats. However, no significant increase in ovarian performance was found under such endocrine and metabolic conditions.

Influence of parity on follicular dynamics and resumption of ovarian cycle in postpartum dairy cows.

The present study examined ovarian changes preceding the resumption of the ovarian cycle in postpartum dairy cows with different parities under similar body nutritional conditions. In postpartum primi- (n=6), bi- (n=4), and multiparous (n=6) Holstein dairy cows, ovarian ultrasonographic observations starting at 7 days after calving were performed every other day and then daily after the confirmation of clinical signs of oestrus for the detection of postpartum first ovulation. Blood samples were collected at the same time as ultrasonography and analyzed for oestradiol and progesterone to monitor ovarian activity. To evaluate the nutritional condition of the cows, body weight and body condition score (BCS, 1=emaciated to 5=obese) were measured weekly and blood samples for the analysis of glucose, insulin, and non-esterified fatty acid (NEFA) were collected at the same time until postpartum second ovulation. Dominant follicles (>8mm in diameter) of the first follicular wave were detected at 7 days after calving in all cows. The first wave follicle ovulated in five of six multiparous cows, whereas no first wave follicle ovulated in any of the primiparous cows. The days to first ovulation after calving in primiparous cows (31.8+/-8.3 days) were significantly greater (p<0.05) than those in multiparous cows (17.3+/-6.3 days), but were not significantly different from biparous cows (28.8+/-8.6 days). There was a significant relationship between parity and days to first ovulation after calving (p<0.05). BCS was maintained at a level of more than 2.5 during the postpartum period in all cows and there was no influence of parity on postpartum changes in BCS, glucose, insulin, or NEFA throughout the experiment. The present study demonstrated a negative relationship between parity and number of days from calving to first ovulation in dairy cows under similar body nutritional conditions. It is possible that the influence of parity on the resumption of ovarian cycle is modulated by the factors different from the nutrition-related changes during the postpartum period in dairy cows.

Changes in the peripheral concentrations of inhibin, follicle-stimulating hormone, luteinizing hormone, progesterone and estradiol-17beta during turnover of cystic follicles in dairy cows with spontaneous follicular cysts.

Several studies have clarified that the follicular cysts degenerate and are replaced by newly growing follicles that develop into new follicular cysts without ovulation, i.e., turnover of ovarian follicular cysts in cows. However, the relativity of endocrinological changes, including the inhibin profile during turnover of spontaneous follicular cysts in dairy cows, is still unclear. In the present study, the relationship between turnover of follicular cysts and changes in the peripheral blood concentrations of progesterone (P), estradiol-17beta (E(2)), luteinizing hormone (LH), follicle stimulating hormone (FSH) and inhibin were examined in lactating dairy cows. Five cows diagnosed with follicular cysts (follicles of more than 25 mm in diameter in the absence of a corpus luteum) were investigated. Their ovarian dynamics were monitored using ultrasonography, and blood samples were collected at 2- or 3- day intervals throughout the experiment. The day when a follicle fated to become a follicular cyst reached more than 8 mm in diameter was defined as the start of a cystic follicular wave. Four of the 5 cows exhibited a similar patterns of cystic follicular changes and hormone profiles. The data from the 4 cows was used for analysis of the relationships between turnover of cystic follicles and the hormone profiles. Two or three new cystic follicular waves occurred in each cow during the experimental period. The mean diameter of the cystic follicles was more than 25 mm 13 to 15 days after the start of the cystic follicular wave, and it began to decrease 1 to 6 days before the start of the subsequent cystic follicular wave. The levels of E(2) and inhibin tended to decrease for 7 to 9 days before the start of a new cystic follicular wave and to increase concomitantly with new follicular cyst growth. The levels of FSH rose for 1 to 3 days before the start of a new cystic follicular wave. The present study clarified the relationship between FSH and inhibin during turnover of spontaneous follicular cysts in dairy cows and found that it was very similar to previous results for cows. The present results suggest that an increase in FSH secretion following a reduction in inhibin secretion triggers turnover of cystic follicles in cows with spontaneous follicular cysts.

Does exogenous progesterone and oestradiol treatment from the mid-luteal phase induce follicular cysts in goats?

The purpose of the present study was to investigate the effects of exogenous ovarian steroid treatment, which is known to induce follicular cyst experimentally in cows, on ovarian activity in goats. Eleven female Shiba goats with the length of the normal oestrous cycle (approximately 21 days) received subcutaneously either 1 ml of ethanol (control group, n=4) or 4 mg of progesterone and 2mg of oestradiol (treatment group, n=7) daily for 7 days beginning on day 14 of the oestrous cycle (day 0=ovulation). Ultrasonographic images of the ovary and blood samples were collected daily to monitor the ovarian activity. Ovulation was observed before 1 day after the end of treatment in the control group. In the treatment group, no detectable structures of follicles or corpus luteum (static ovarian condition) were found for 6.0+/-1.4 days (mean+/-S.D.) after the end of treatment. Then, detectable follicles appeared and ovulation was observed in all animals of the treatment group. There was no significant difference in the maximum diameter of the ovulatory follicle between the control and treatment group (4.7+/-0.4mm versus 5.1+/-0.7 mm). The large non-ovulatory follicles, which grew more than 10mm in diameter were observed after the static ovarian condition in one goat of the treatment group, whereas no turnover of the cystic follicular structures was found. The length of the inter-ovulatory intervals in the treatment group was significantly longer than that in the control group (38.4+/-7.4 days versus 20.3+/-0.5 days, P<0.05). The present results demonstrated that the exogenous treatment of progesterone and oestradiol, which was adapted from the follicular cyst model in cows, did not induce follicular cysts in goats, suggesting that there is/are different mechanism(s) mediating the occurrence of follicular cysts between cows and goats.

Changes in the ovarian dynamics and endocrine profiles in goats treated with a progesterone antagonist during the early luteal phase of the estrous cycle.

The aim of the present study was to determine the physiological role of endogenous progesterone in the regulation of ovarian dynamics, gonadotropin and progesterone secretion during the early luteal phase in the goat. Cycling Shiba goats received subcutaneously a vehicle (control group, n=5) or 50 mg of RU486 (RU486 group, n=4) daily from 1 to 7 days after ovulation (day 0) determined by transrectal ultrasonography. Ovarian dynamics were monitored by the ultrasonography and blood samples were collected daily until the subsequent ovulation for analysis of progesterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion. Blood samples were also collected at 10 min intervals for 6 h on day 3 and day 7 for the analysis of pulsatile patterns of LH and FSH. The LH pulse frequency was significantly (P<0.05) higher in the RU486 group than in the control group on day 7 (4.8+/-1.1 pulses/6 h versus 1.2+/-0.4 pulses/6 h). The shape of the FSH pulses was unclear on day 3 and day 7 in both groups and the overall means of FSH concentration for 6 h on day 3 and day 7 were not significantly different between the RU486 and the control groups. The pattern of daily FSH concentrations showed a wave-like fluctuation in both groups. There was no significant difference in the inter-peak intervals of the wave-like pattern of daily FSH secretion between the RU486 and the control groups (4.1+/-0.6 days versus 4.5+/-0.6 days). The maximum diameter of the largest follicle that grew from day 1 to day 7 in the RU486 group tended to be greater than that in control goats (6.4+/-0.8 mm versus 5.0+/-0.8 mm, P=0.050), whereas no significant difference was detected in the size of the corpus luteum and progesterone concentrations between the control and RU486 groups on almost all days during the treatment period. These results indicate that the rise of the progesterone concentration suppresses the pulsatile LH secretion and follicular growth, whereas progesterone has no physiological role in the regulation of FSH secretion and luteal function during the early luteal phase of the estrous cycle in goats.

Estrus synchronization and conception rate after a progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in heifers.

The objective of the present study was to evaluate estrus synchronization and conception rate after progesterone releasing intravaginal device (PRID) treatment from the early luteal phase in the presence or absence of estradiol benzoate (EB) in heifers. Heifers (n=11) were assigned randomly to two treatments; insertion of a PRID containing 1.55 g progesterone with a capsule attached including 10 mg EB (P+EB; n=6) and the PRID withdrawn the EB capsule (P-EB; n=5). The PRID was inserted into the vagina on Day 2 of the estrous cycle (Day 0 was the day of ovulation) and was left for 12 days. The proportion of heifers exhibiting standing estrus within 3 days after PRID removal was 83.3% (5/6) for the P+EB group, and 80.0% (4/5) for the P-EB group, respectively. Conception rate by artificial insemination on synchronized estrus was 80.0% (4/5) in the P+EB group, and 100% (4/4) in the P-EB treatment group, respectively. These results suggest that a PRID treatment from 2 days after ovulation for 12 days in the presence or absence of EB has an effect on the synchronization of estrus and produces a beneficial conception rate in heifers.

Ovarian and hormonal responses to a progesterone-releasing controlled internal drug releasing treatment in dietary-restricted goats.

An experiment was conducted to evaluate the effects of dietary restriction on ovarian, endocrine (ovarian steroids and luteinizing hormone (LH) pulse) and metabolic (glucose, insulin and non-esterified fatty acid (NEFA)) profiles in goats treated with a progesterone-releasing controlled internal drug releasing (CIDR-G) device. Cycling goats were offered either a maintenance or a restricted (30% of requirement; n =4 per treatment) level of feeding. The dietary restriction was started on the day following ovulation. At 30-32 days after the start of food restriction, the goats received a prostaglandin F(2alpha) (2mg of dinoprost) injection followed by 10 days of CIDR-G treatment. Ovarian ultrasonographic images were monitored daily throughout the experiment and blood samples were collected daily just before the morning feeding for analysis of endocrine and metabolic profiles. Frequent blood samples (1 ml) were also collected at 10 min intervals for 8 h from -8 h to CIDR-G removal, and from 32 to 40 h after CIDR-G removal for analysis of LH pulses. Body weight was significantly (P < 0.05) decreased in the food-restricted animals. Oestrous behaviour and ovulation followed by a rise of plasma progesterone concentration were observed after the CIDR-G removal in all control animals but not in any of the food-restricted animals within 12 days after CIDR-G removal. The LH pulse frequency from 32 to 40 h after the CIDR-G removal was significantly (P < 0.05) lower in the food-restricted animals than in control animals (1.5 +/- 0.6 versus 3.8 +/- 0.5 pulses for 8 h). There was no significant difference in the glucose concentration in weekly plasma samples between control and food-restricted animals. Insulin concentrations from 2 weeks after the start of feed restriction were significantly (P < 0.05) lower in restricted animals than in control animals. The NEFA concentration in restricted animals was significantly (P < 0.05) increased after the start of feed restriction, and then decreased gradually to the basal level. The present results suggest that nutritionally induced anovulation after CIDR-G treatment is associated with a reduction in the frequency of LH pulses, and that insulin and NEFA, rather than the glucose concentration in the circulation, may be associated with the metabolic suppression of LH pulses.