Estrus induction with prostaglandin F2alpha, cloprostenol or fenprostalene during the normal estrous cycle, superovulation and after embryo collection

Holstein heifers used as embryo donors were treated with three luteolytic agents (PGF2alpha, cloprostenol, fenprostalene) during the normal estrous cycle, superovulation or after embryo collection to determine the interval from treatment to estrus. A similar return-to-estrus interval was observed for each luteolytic agent among the three groups of heifers. Nevertheless, after embryo collection, fenprostalene had a tendency to induce the longest delays (p = 0.08). This tendency is supported by a higher proportion of delayed luteolysis and more heifers showing estrus later than 11 d post treatment. Also, during normal estrous cycles, 5/10 and 0/8 fenprostalene- and cloprostenol-treated heifers, respectively, showed progesterone concentrations higher than 1 ng/mL 48 h after treatment. Regardless of the luteolytic agent used, estrus was induced earlier (P < 0.005) during superovulation than when heifers were treated between Days 9 to 16 of the normal estrous cycle or after embryo collection. However, the return-to-estrus interval was similar between heifers treated during superovulation and those treated between Days 6 to 8 of the normal estrous cycle. After embryo collection, intervals before the return to estrus increased with the number of Corpora lutea (CL) palpated except in the nonresponding group (0 to 1 CL), which returned to estrus later than the low responding group (2 to 4 CL).     

Nonsurgical embryo collection from sows with surgically shortened uteri

To allow for the nonsurgical collection of swine embryos, the uteri of sows (n=7) were surgically shortened. A section of each uterine horn was resected to facilitate a transcervical flushing procedure. All sows with a shortened uterus exhibited natural estrus at least once after the operation. Four to six days after insemination, embryos were collected with a two-way Foley catheter. Embryos were collected (n=55, 6.3+/-6.0: x +/-SD ) from every treated sow. Although treated sows often did not exhibit estrus beyond 1 to 9 natural estruses, those sows (n=27) with persistent corpora lutea (CL) over a 4 to 5 wk period were given prostaglandin F(2alpha) (PGF(2alpha)) and they returned to estrus in 5.2+/-1.1 d: x +/-SD .     

Do high progesterone concentrations decrease pregnancy rates in embryo recipients synchronized with PGF2alpha and eCG?

The objective of this study was to evaluate the effects of equine chorionic gonadotropin (eCG) treatment on the number of induced accessory corpora lutea (CL), plasma progesterone concentrations and pregnancy rate in cross-bred heifers after transfer of frozen-thawed (1.5M ethylene glycol) embryos. All recipients received 500 microg PGF2alpha (dl-cloprostenol, i.m.) at random stages of the estrous cycle (Day 0) and were observed for estrus for 7 days. On Day 14, heifers detected in estrus between 2 and 7 days after PGF2alpha treatment were randomly allocated to four groups ( n=83 per group) and given 0 (control), 200, 400, or 600 IU of eCG. Two days later (Day 16), these recipients were given PGF2alpha and observed for estrus. Six to eight days after detection of estrus, plasma samples were collected to determine progesterone concentration and ultrasonography was performed to observe ovarian structures. Heifers with multiple CL or a single CL >15 mm in diameter received an embryo by direct transfer. Embryos of excellent and good quality were thawed and transferred to the recipients by the same veterinarian. Pregnancy was diagnosed by ultrasonography and confirmed by transrectal palpation 21 and 83 days after embryo transfer (ET), respectively. Plasma progesterone concentrations on the day of transfer (Day 7 of the estrous cycle) were 3.9+/-0.7, 4.2+/-0.4,6.0+/-0.4 and 7.8+/-0.6 ng/ml for groups Control, 200, 400, and 600, respectively (Control versus treated groups P=0.009; 200 versus 400 and 600 groups P=0.0001; and 400 versus 600 P=0.012 ). Conception rates 83 days after ET were 41.9, 50.0, 25.0, and 20.9% for groups Control, 200, 400, and 600, respectively (200 versus 400 and 600 groups P=0.0036 ). In conclusion, an increase in progesterone concentration, induced by eCG treatment, did not improve pregnancy rates in ET recipients. Conversely, there was a decline in conception rates in the animals with the highest plasma progesterone concentrations.     

Ultrasound and endocrine evaluation of the ovarian response to PGF2alpha given at different stages of the luteal phase in ewes

The purpose of this study was to evaluate the ovarian response of ewes to two treatments with PGF2alpha using transrectal ovarian ultrasonography and hormone measurements. Fifteen milligrams of PGF2alpha was given to six cyclic Western White Face (WWF) ewes early in the estrous cycle (Days 4 to 7) and to six late in the cycle (Days 10 to 12 after ovulation), and a second treatment was given 9 days after the first. Ultrasound scanning and blood sampling started 7 days prior to the first PGF2alpha treatment and ended 10 days (scanning) or 19 days (blood sampling) after the second PGF2alpha treatment, for both groups of ewes. Mean ovulation rate (2.6 +/- 0.7) did not differ significantly between the ewes first treated early or late in the cycle, or after the first or second treatments with PGF2alpha. The time from treatment to ovulation was longer in ewes first treated early (4.0 +/- 0.3 days) compared to late (2.8 +/- 0.4 days) in the cycle (P < 0.05). Both the number of ovulations (range: 0-7) and time from treatment to ovulation (range: 1-9 days) were highly variable. This variability appeared to be due to the extension of the life span of ovulating follicles that emerged prior to PGF2alpha administration and also ovulation of some follicles that emerged after treatment. When results for first and second treatments were pooled, the total number of follicles > 5 mm in diameter on the day of treatment that failed to ovulate in response to PGF2alpha was higher in ewes first treated early (0.8 +/- 0.2/ewe) compared to late (0.3 +/- 0.2/ewe) in the cycle (P < 0.05). The proportion of detected luteal structures relative to the number of ovulations was lower in ewes first treated early compared to late in the cycle (60 and 86%, respectively; P < 0.05). Disruption of ovulatory follicle dynamics and normal luteogenesis, and variability in the timing of ovulation after PGF2alpha treatments could all contribute to poor or variable fertility when prostaglandins are used for estrus synchronization.     

Follicular growth, ovulation and embryo recovery in dairy cows given FSH at the beginning or middle of the estrous cycle

Variability in the superovulation response is an important problem for the embryo transfer industry. The objective of this study was to determine whether FSH treatment at the beginning of the cycle would improve the ovulation rate and embryo yield in dairy cows. Twenty-eight postpartum cyclic dairy cows were allocated at random to 4 treatment groups (A, B, C and D). Group A cows (n = 10) received FSH (35 mg) at a decreasing dose, starting on Day 9 (Day 0 = day of estrus) for 5 days followed by PGF(2alpha) (35 mg) on Day 12. Cows assigned to Groups B, C and D (n = 6 cows each, respectively) were given 35 mg FSH at a decreasing dose from Days 2 to 6 followed by PGF(2alpha) on Day 7. Group C and D cows received PRID inserts from Day 3 to Day 7. Cows in Group D additionally received 1000 IU hCG 60 hours after PGF(2alpha) treatment. Ovaries were scanned daily using a real time ultrasound scanner from the beginning of FSH treatment until embryo recovery, to monitor follicular development, ovulation and the number of unovulated follicles. Embryos were recovered from the uterus by a nonsurgical flushing technique 7 days after breeding. There were no differences (P>0.01) in the number of follicles > 10 mm at 48 hours after PGF(2alpha) treatment among the 4 groups. The mean numbers of follicles were 10.6 +/- 1.2, 9.3 +/- 1.3, 12.2 +/- 1.3 and 15.0 +/- 2.9 for Groups A, B, C and D, respectively. A significantly (P<0.001) higher number of ovulations was observed and a larger number of embryos was recovered in Group A than in the other groups. The results of this study indicate that superovulation with FSH at the beginning of the cycle causes sufficient follicular development but results in very low ovulation and embryo recovery rates.     

Simultaneous injection of PGF2alpha and GnRH into diestrous dairy cows delays return to estrus

Simultaneous injections of prostaglandin F2alpha (PGF) and gonadotropin releasing hormone (GnRH) or saline were given to 32 diestrous dairy cows to test the ability of GnRH to improve estrous and ovulation synchrony beyond that of PGF alone. Cows were randomly assigned to receive PGF on Day 8 or Day 10 of the estrous cycle (estrus = Day 0), and all cows were further assigned to simultaneous injection of GnRH or saline. Corpus luteum (CL) regression, return to estrus and follicular activity were monitored by plasma progesterone assay, twice-daily estrous detection and ultrasonographic examination, respectively. Plasma progesterone concentrations declined to <1.0 ng/ml at 24 hours after PGF in all cows and were not affected by GnRH. Gonadotropin releasing hormone inducted premature ovulation or delayed return to estrus in 7 of 8 cows treated with PGF/GnRH on Day 8 and 3 of 8 cows treated with PGF/GnRH on Day 10. Further, cows with premature GnRH-induced ovulations failed to develop and maintain a fully functional CL, and all returned to estrus 7 to 13 days after the induced ovulation. These data indicate that GnRH administered simultaneously with a luteolytic dose of PGF disrupts follicular dynamics and induces premature ovulation or delays normal return to estrus and, therefore, does not improve the synchrony of estrus and ovulation achieved with PGF alone.     

Ovarian and endocrine responses to prostaglandin F2alpha (PGF2alpha) given at the expected time of the endogenous FSH peak at mid-cycle in ewes

In the ewe, a rise in circulating concentrations of FSH preceding follicular wave emergence begins in the presence of growing follicles from a previous wave. We hypothesized that prostaglandin F(2alpha) (PGF(2alpha)) given at the time of an endogenous FSH peak in cyclic ewes would result in synchronous ovulation of follicles from two consecutive waves, increasing ovulation rate. Twelve Western White Face (WWF) ewes received a single i.m. injection of PGF(2alpha) (15 mg/ewe) at the expected time of a peak in FSH secretion, from Days 9 to 12 after ovulation. The mean ovulation rate after PGF(2alpha) treatment (2.3+/-0.3) did not differ (P>0.05) from the pre-treatment ovulation rate (1.7+/-0.1). Five ewes ovulated follicles from follicular waves emerging before and after PGF(2alpha) injection (3.0+/-0.6 ovulations/ewe) and seven ewes ovulated follicles only from a wave(s) emerging before PGF(2alpha) treatment (2.0+/-0.3 ovulations/ewe; P>0.05). The mean interval from PGF(2alpha) to emergence of the next follicular wave (1.0+/-0.4 and 4.0+/-0.0 d, respectively; P<0.001) and the interval from PGF(2alpha) treatment to the next FSH peak (0 and 3.5+/-0.4d, respectively; P<0.05) differed between the two groups. Six ewes ovulated after the onset of behavioral estrus, with a mean ovulation rate of 1.7+/-0.2, and six ewes ovulated both before and after the onset of estrus (3.0+/-0.5 ovulations/ewe; P<0.05). None of the ovulations that occurred before estrus resulted in corpora lutea (CL) with a full life span. At 24h before ovulation, follicles ovulating before or after the onset of estrus differed in size (4.1+/-0.3 or 5.5+/-0.4mm, respectively; P<0.05) and had distinctive echotextural characteristics. In conclusion, the administration of PGF(2alpha) at the expected time of an FSH peak at mid-cycle in ewes may alter the endogenous rhythm of FSH secretion and was not consistently followed by ovulation of follicles from two follicular waves. In non-prolific WWF ewes, PGF(2alpha)-induced luteolysis disrupted the normal distribution of the source of ovulatory follicles and may be associated with untimely follicular rupture and luteal inadequacy.     

Impact of pre-ovulatory follicle diameter on plasma estradiol, subsequent luteal profiles and conception rate in buffalo (Bubalus bubalis)

The present study was designed to investigate the impact of pre-ovulatory follicle (POF) diameter on the day of estrus on plasma estradiol concentration, subsequent luteal profile (corpus luteum, CL, diameter and plasma progesterone concentration) and conception rate in buffaloes. Twenty-eight buffaloes were synchronized with synthetic analogue of prostaglandin F(2α) (PGF(2α)) administered 11 days apart. Transrectal ultrasonography and jugular vein blood sampling was carried out on the day of estrus and on days 0 (day of ovulation), 5, 12, 16 and 21 post-ovulation. Out of 28 buffaloes, 11 (39.3%) were diagnosed pregnant on day 40 post-ovulation. Retrospective analysis revealed that the buffaloes getting pregnant had larger (p<0.05) POF diameter. In fact, all the buffaloes (n=5/5) having POF diameter between >14 and 16 mm conceived, whereas, conception rate in buffaloes with POF diameter between >12 and ≤14 mm (n=6/17) or <12 mm (n=0/6) was 35.3% and 0.0%, respectively. A positive correlation (r=0.57, p<0.05) was observed between POF diameter and plasma estradiol concentration at estrus. On day 5 post-ovulation, luteal profile was positively correlated (CL: r=0.34, p<0.05; plasma progesterone concentration: r=0.27, p>0.05) with POF diameter. Further, on the same day, plasma progesterone concentration was positively correlated (r=0.47, p<0.05) with CL diameter, however, this correlation was absent (r=0.05, p>0.05) during the subsequent luteal phase. Nevertheless, the post-ovulation luteal profile of pregnant buffaloes was higher (p<0.05) compared to non-pregnant counterparts. In conclusion, the diameter of POF in buffaloes has positive impact on plasma estradiol concentration at estrus, post-ovulation luteal profile and conception rate. The diameter of CL can be used as an indicator of luteal function at early but not at mid or late luteal phase of estrus cycle in buffaloes.     

Is slow follicular growth the cause of silent estrus in water buffaloes?

The present experiment was conducted to study the growth profile of the ovulatory follicle in relation to the expression of estrus following administration of PGF(2alpha) to subestrus buffaloes. After detection of a mature corpus luteum by examination per rectum, confirmed by ultrasound scanning, subestrus buffaloes (n=20) were treated (Day 0) with single dose of Dinoprost tromethamin (25 mg, i.m.). Blood samples were collected at 0, 24 and 48 h after treatment for estimation of plasma progesterone concentration. Growth profile of the ovulatory follicle was monitored daily through ultrasound scanning starting from Day 0 until ovulation and the regression profile of CL was monitored at 0, 24 and 48 h of treatment. Estrus was detected by exposure to a fertile buffalo bull three times a day until expression of overt estrus or ovulation. Behavioral estrus was recorded in 14 animals and 6 animals ovulated silently. Sixteen animals including six animals with silent estrus ovulated from the dominant follicle present at treatment (Group A) and remaining four animals ovulated from the dominant follicle of succeeding follicular wave (Group B). The intervals from treatment to estrus (6.5+/-0.25 versus 3.2+/-0.27 days, P<0.001) and treatment to ovulation (7.5+/-0.25 versus 5.4+/-0.46 days, P<0.005) were significantly longer in animals of Group B compared with animals of Group A. Significant differences were observed in growth profile of the ovulatory follicle between animals of Groups A and B with respect to size of the follicle on Day 0 (9.8+/-0.7 versus 5.3+/-0.45 mm, P<0.001), daily growth rate (0.97+/-0.07 versus 1.6+/-0.2 mm/day, P<0.01) and increase in diameter (4.1+/-0.6 versus 7.8+/-0.7 mm, P<0.01). The animals with silent estrus (subgroup A-2) had significantly smaller diameter of the ovulatory follicle on Day 0 (7.7+/-0.4 versus 11.0+/-0.7 mm, P<0.005), its daily growth rate was significantly slower (0.7+/-0.02 versus 1.1+/-0.1 mm/day, P<0.01) and they recorded significantly longer interval from treatment to ovulation (7.3+/-0.56 versus 4.2+/-0.27 days, P<0.001) compared with the animals that showed overt estrus (subgroup A-1). The corpus luteum area (CL area) and plasma progesterone (P(4)) concentration declined continuously from 0 to 48 h after PGF(2alpha) treatment in the animals of both the Groups A and B. Non-significant differences were observed in mean CL area and plasma P(4) concentration at 0, 24 and 48 h post-treatment between animals of Groups A and B and also between animals of subgroups A-1 and A-2. The small size and the slow growth rate of the ovulatory follicle were identified as the possible cause of silent estrus in subestrus buffaloes after PGF(2alpha) treatment.     

Environmental,genetic and social factors affecting the expression of estrus in beef cows

Genetic, social and environmental factors affecting behavioral estrus were evaluated in Angus (n = 10), Brahman (n = 10) and Senepol (n = 10) cows during a PGF2alpha synchronized estrus and subsequent spontaneous estrus. Cows were equally stratified by breed to two groups of 15. Both groups were pre-synchronized with a modified two-injection PGF2alpha protocol. At the start of the experiment, cows were treated with 25 mg PGF2alpha followed by a second and third administration of 12.5 mg PGF2alpha, 11 and 12 days later to induce synchronized estrus. The subsequent estrus was designated as spontaneous estrus. Behavioral estrus data including the onset and end of estrus, estrous duration and the total number of mounts received for the synchronized and spontaneous estruses were collected using HeatWatch". Interval from the third PGF2alpha, treatment to the onset of a HeatWatch" estrus occurred earlier (P < 0.05) in Angus (31 +/- 5 h) than Brahman (53 +/- 7 h) or Senepol (53 +/- 4 h) cows, with dominant Senepol and Brahman cows taking longer to exhibit estrus after PGF2alpha than subordinate cows. The duration of the synchronized estrus tended to be shorter (P < 0.06) in Senepol (12 +/- 3 h) than in Angus (19 +/- 2 h) or Brahman (17 +/- 2 h) cows. Behavioral estrus data between the two periods were confounded by greater temperature-humidity index (THI) values during spontaneous estrus. The THI during spontaneous estrus appeared (P = 0.09) to affect the duration of estrus (9 +/- 1 h versus 16 +/- 1 h) and did affect (P < 0.0001) the total number of mounts received (8 +/- 4 mounts versus 34 +/- 4 mounts) during spontaneous estrus compared to synchronized estrus. Breed had no effect (P > 0.10) on the duration and total number of mounts received during synchronized and spontaneous estruses. In conclusion, type of estrus (synchronized or spontaneous), THI, social dominance and breed exerted significant effects on characteristics associated with behavioral estrus in beef cattle in subtropical environments.     

Regulation of estrus and ovulation in mares with progesterone or progesterone and estradiol biodegradable microspheres with or without PGF2alpha

A single injection of a microsphere preparation, designed to deliver 1.25 gm progesterone and 100 mg estradiol-17beta at a controlled rate, for a duration of 12 to 14 days, produces accurate control of estrus and fertile ovulations in mares. Theatment is followed by PGF(2)alpha injection 14 days after steroid injection. The objectives of the present study were to determine whether estradiol added to the progesterone treatment or PGF(2)alpha administered at the end of the steroid treatment regimen, would improve synchronization of estrus and ovulation. A total of 45 cyclic horse mares was randomly assigned to 1 of 5 treatment groups as follows: Group 1 (control, n=9) sterile microsphere vehicle + sterile PGF(2)alpha vehicle 14 days after treatment with microsphere vehicle; Group 2 (n=9) progesterone and estradiol microspheres + PGF(2)alpha 14 days after treatment with microspheres; Group 3 (n=9) progesterone and estradiol microspheres + PGF(2)alpha vehicle 14 days after treatment with microspheres; Group 4 (n=9) progesterone + PGF(2)alpha 14 days after treatment with microspheres; and Group 5 (n=9) progesterone + PGF(2)alpha vehicle 14 days after treatment with microspheres. Addition of estradiol (P<0.05) or PGF(2)alpha (P<0.05) to the treatment regimen increased synchronization efficary by reducing variation in days to ovulation. All treatments significantly reduced variation in days to estrus compared with that of the controls; however, mares in the progesterone groups had an increased incidence of silent or shortened estrous behavior (<- 2 days) following treatment. Estradiol added to the treatment regimen increased (P<0.05) the number of mares with post treatment estrus > 2 days in duration compared with mares treated with progesterone (78 vs 33%, respectively). Therefore, estradiol and PGF(2)alpha each appear to reduce variation in days to ovulation while estradiol seems to promote better expression of posttreatment estrous behavior.     

The effects of low level of feeding on response to synchronization of estrus, ovulation rate and embryo loss in goats

Mature nonlactating British Saanen and Toggenburg does with a body score 2 were fed 25% (n=24) and 100% (n=16) maintenance rations from about 19 days before mating until slaughter at approximately 60 days after mating. Estrus was synchronized using PGF(2alpha), and the ovulation rate was determined by laparoscopic examination of the ovaries once between Days 6 and 10 after mating. Pregnancy rate, potential kidding rate and embryo loss were determined by counts of viable fetuses at slaughter. The proportion of does in estrus within 96 hours of PGF(2alpha) administration was not different (P<0.5) between the feed-restricted and the maintenance groups (71.0% and 87.5%, respectively); however, the time of onset of estrus after PGF(2alpha) tended to be longer (P=0.12) in the feed-restricted group. Ovulation rate, incidence of multiple ovulations and proportion of does pregnant at 60 days were significantly lower (P=0.0004, P=0.025, P=0.05, respectively) in the restricted group. More embryos from single than multiple ovulations were lost in the restricted group (P=0.01). There was no difference in the overall ovulatory activity between right and left ovaries in the 2 groups. Transuterine migrations were observed in all does that had unilateral multiple ovulations. No migration was observed in does which had single ovulations. These data indicate that restricted feed intake in goats tended to delay the onset of estrus and lowered the ovulation rate, incidence of multiple ovulations, and pregnancy rate.     

Follicular waves during the oestrous cycle in Nili-Ravi buffaloes undergoing spontaneous and PGF2alpha-induced luteolysis

The objective of this study was to characterize the follicular waves and associated ovarian events during spontaneous and PGF(2alpha)-induced oestrous cycles in Nili-Ravi buffaloes. In Exp. 1, (n=13 oestrous cycles) follicular and luteal development was monitored by ultrasonography and jugular blood samples were collected simultaneously on alternate days. Of 12 oestrous cycles, 9 (75%) had two waves of follicular activity and only 3 (25%) had three waves. The mean (+/-S.E.M.) length of the oestrous cycle was shorter (P<0.05) in buffaloes with two waves than in those with three waves (21.2+/-0.1 days versus 22.8+/-0.1 days). In Exp. 2, follicular dynamics were compared in buffaloes undergoing spontaneous (n=12 oestrous cycles) and PGF(2alpha)-induced (n=9) regression of the corpus luteum (CL). The dynamics of ovulatory follicular growth during the 3 days before oestrus were similar (P>0.05) in buffaloes undergoing spontaneous and PGF(2alpha)-induced luteolysis. These results show that (1) the majority of buffaloes had a two wave pattern of follicular growth and emergence of a third wave was associated with a longer luteal phase, and (2) follicular dynamics during the 3 days before oestrus were similar in buffaloes undergoing spontaneous and PGF(2alpha)-induced luteolysis.     

Evaluating recipient and embryo factors that affect pregnancy rates of embryo transfer in beef cattle

The objectives of this experiment were to determine the effects of corpus luteum characteristics, progesterone concentration, donor-recipient synchrony, embryo quality, type, and developmental stage on pregnancy rates after embryo transfer. We synchronized 763 potential recipients for estrus using one of two synchronization protocols: two doses of PGF2alpha (25 mg i.m.) given 11 d apart (Location 1); and, a single norgestomet implant for 7 d with one dose of PGF2alpha (25 mg i.m.) 24 h before implant removal (Location 2). At embryo transfer, ovaries were examined by rectal palpation and ultrasonography. Of the 526 recipients presented for embryo transfer, 122 received a fresh embryo and 326 received a frozen embryo. Pregnancy rates were greater (P < 0.05) with fresh embryos (83%) than frozen-thawed embryos (69%). Pregnancy rates were not affected by embryo grade, embryo stage, donor-recipient synchrony, or the palpated integrity of the CL. Corpus luteum diameter and luteal tissue volume increased as days post-estrus for the recipients increased. However, pregnancy rates did not differ among recipients receiving embryos 6.5 to 8.5 days after estrus (P > 0.1). There was a significant, positive simple correlation between CL diameter or luteal tissue volume and plasma progesterone concentration (r = 0.15, P < 0.01 and r = 0.18, P < 0.01, respectively). There were no significant differences in mean CL diameter, luteal volume or plasma progesterone concentration among recipients that did or did not become pregnant after embryo transfer. We conclude that suitability of a potential embryo transfer recipient is determined by observed estrus and a palpable corpus luteum, regardless of size or quality.     

Control of ovulation with a GnRH agonist after superstimulation of follicular growth in buffalo: fertilization and embryo recovery

The potential to use a GnRH agonist bioimplant and injection of exogenous LH to control the time of ovulation in a multiple ovulation and embryo transfer (MOET) protocol was examined in buffalo. Mixed-parity buffalo (Bubalus bubalis; 4-15-year-old; 529 +/- 13 kg LW) were randomly assigned to one of five groups (n = 6): Group 1, conventional MOET protocol; Group 2, conventional MOET with 12 h delay in injection of PGF2alpha; Group 3, implanted with GnRH agonist to block the preovulatory surge release of LH; Group 4, implanted with GnRH agonist and injected with exogenous LH (Lutropin, 25 mg) 24 h after 4 days of superstimulation with FSH; Group 5, implanted with GnRH agonist and injected with LH 36 h after superstimulation with FSH. Ovarian follicular growth in all buffaloes was stimulated by treatment with FSH (Folltropin-V, 200 mg) administered over 4 days, and was monitored by ovarian ultrasonography. At the time of estrus, the number of follicles >8 mm was greater (P < 0.05) for buffaloes in Group 2 (12.8) than for buffaloes in Groups 1(8.5), 3 (7.3), 4 (6.1) and 5 (6.8), which did not differ. All buffaloes were mated by Al after spontaneous (Groups 1-3) or induced (Groups 4 and 5) ovulation. The respective number of buffalo that ovulated, number of corpora lutea, ovulation rate (%), and embryos + oocytes recovered were: Group 1 (2, 1.8 +/- 1.6, 18.0 +/- 13.6, 0.2 +/- 0.2); Group 2 (4,6.1 +/- 2.9, 40.5 +/- 17.5, 3.7 +/- 2.1); Group 3 (0, 0, 0, 0); Group4 (6, 4.3 +/- 1.2, 69.3 +/- 14.2, 2.0 +/- 0.9); and Group 5 (1, 2.5 +/- 2.5, 15.5 +/- 15.5, 2.1 +/- 2.1). All buffaloes in Group 4 ovulated after injection of LH and had a relatively high ovulation rate (69%) and embryo recovery (46%). It has been shown that the GnRH agonist-LH protocol can be used to improve the efficiency of MOET in buffalo.     

Does injection of prostaglandin F(2alpha) (PGF2alpha) cause ovulation in anestrous Western White Face ewes?

In a previous study in our laboratory, treatment of non-prolific Western White Face (WWF) ewes with PGF(2 alpha) and intravaginal sponges containing medroxyprogesterone acetate (MAP) on approximately Day 8 of a cycle (Day 0 = first ovulation of the interovulatory interval) resulted in ovulations during the subsequent 6 days when MAP sponges were in place. Two experiments were performed on WWF ewes during anestrus to allow us to independently examine if such ovulations were due to the direct effects of PGF(2 alpha) on the ovary or to the effects of a rapid decrease in serum concentrations of progesterone at PGF(2 alpha)-induced luteolysis. Experiment 1: ewes fitted with MAP sponges for 6 days (n = 12) were injected with PGF(2 alpha) (n = 6; 15 mg im), or saline (n = 6) on the day of sponge insertion. Experiment 2: ewes received progesterone-releasing subcutaneous implants (n = 6) or empty implants (n = 5) for 5 days. Six hours prior to implant removal, all ewes received a MAP sponge, which remained in place for 6 days. Ewes from both experiments underwent ovarian ultrasonography and blood sampling once daily for 6 days before and twice daily for 6 days after sponge insertion. Additional blood samples were collected every 4 h during sponge treatment. Experiment 1: 4-6 (67%) PGF(2 alpha)-treated ewes ovulated approximately 1.5 days after PGF(2 alpha) injection; these ovulations were not preceded by estrus or a preovulatory surge release of LH, and resulted in transient corpora hemorrhagica (CH). The growth phase was longer (P < 0.05) and the growth rate slower (P < 0.05) in ovulating versus non-ovulating follicles in PGF(2 alpha)-treated ewes. Experiment 2: in ewes given progesterone implants, serum progesterone concentrations reached a peak (1.7 2 ng/mL; P < 0.001) on the day of implant removal and decreased to basal concentrations (<0.17 ng/mL; P < 0.001) within 24 h of implant removal. No ovulations occurred in either the treated or the control ewes. We concluded that ovulations occurring after PGF(2 alpha) injection, in the presence of a MAP sponge, could be due to a direct effect of PGF(2 alpha) at the ovarian level, rather than a sudden decline in circulating progesterone concentrations.     

Progesterone concentration as an indicator of ovarian response to superovulation in Chios ewes

We investigated the prediction of the ovarian response to superovulation using progesterone (P4) determination in Chios ewes. During the estrus period. estrus synchronization and multiple ovulations were induced in 100 non-pregnant, non-lactating Chios ewes by a combination of FGA-impregnated intravaginal sponges and 8.8 mg of ovine FSH. Laparoscopic insemination was conducted 24-28 h after the onset of estrus. A concentration of P4 was determined on Day 5 of the estrous cycle and on Day 6 the ovarian response was evaluated by counting the corpus lutea (CL); subsequently, embryo collection was performed. According to the response of their ovaries, ewes were allocated into four groups: A (n = 30); B (n = 37); C (n = 22); D (n = 11), with minimal (0-3 CL), moderate (4-8 CL), good (9-13 CL) or extreme (> 13 CL) ovarian response, respectively. In groups C and D, the mean blood serum P4 concentration (23.2 and 27.3 ng/ml, respectively) was higher (P < 0.001) than that in groups A and B (4.6 and 13.1 ng/ml, respectively); no difference was detected in blood P4 concentration between groups C and D. A strong linear relation (F < 0.00005) was found between blood P4 concentration and the number of CL, as well as between blood P4 and a dummy variable corresponding to poor (< 4 CL) or moderate/good/extreme ovarian response (>3 CL). Our results indicate that based on blood P4 measurement, it is feasible to identify ewes that should show the highest embryo recovery, while it is impossible to predict the exact number of CL formed.     

Effect of prostaglandin F2alpha at the time of AI on progesterone levels and pregnancy rate in synchronized Italian Mediterranean buffaloes

The aims of this study were to evaluate the effects of an intravenous or intramuscular PGF2alpha analogue administration on the day of estrus on progesterone concentration and pregnancy rate in buffaloes undergoing artificial insemination (AI). To this end, two experiments were carried out. The first study was performed on 72 Mediterranean buffaloes synchronized by the Ovsynch-TAI Program. On the day of estrus only animals considered in heat were divided into four groups: Groups IVC and IMC received, respectively, an intravenous or intramuscular injection of cloprostenol (0.524 mg), whereas control Groups IVS and IMS received the same injections of saline. Milk samples were collected daily from each animal to assess progesterone concentration in the whey by RIA method. In addition on alternate days, buffaloes underwent transrectal ultrasound analysis. The second study was carried out on 385 buffaloes synchronized by the Ovsynch-TAI Program. On the day of AI, animals were divided in four groups, as described in experiment 1. Pregnancy rate was evaluated either on day 26 or day 45 and embryonic mortality rate was recorded. Statistical analysis was performed by ANOVA and chi2 test. A higher (P<0.05) progesterone concentration was recorded on day 11 (Day 0=estrus day) in Groups IVC and IMC compared to Groups IVS and IMS (351.6+/-129.7 and 355.8+/-112.2 pg ml(-1) vs. 239.8+/-81.1 and 243.6+/-90.5 pg ml(-1), respectively). Furthermore, a larger CL was recorded on the same day in treated vs. control groups (1.25+/-0.15 and 1.27+/-0.17 cm, respectively, in Groups IVC and IMC vs. 1.08+/-0.14 and 1.05+/-0.13 cm in IVS and IMS). In the second study, a higher pregnancy rate was observed in treated (IVC+IMC) vs. control (IVS+IMS) groups (46.7% vs. 30.7%; P<0.01), while no differences were recorded between treated groups. From these data, it can be concluded that either intravenous or intramuscular administration of PGF2alpha at the time of AI can enhance progesterone levels and pregnancy rate in buffaloes.     

Efficacy of either a single or split treatment of PGF2alpha after a 14 day melengestrol acetate treatment to synchronize estrus and induce luteolysis in Bos indicus x Bos taurus heifers

Two experiments evaluated a modified delivery of prostaglandin F2alpha (PGF2alpha) after a melengestrol acetate (MGA) treatment in Angus and Bos indicus x Bos taurus (BI) heifers. Experiment 1 was replicated three times with yearling BI heifers (n = 695). Heifers received MGA (0.5 mg head(-1) day(-1)) for 14 days. In Replications 1 and 2, heifers received either 25 mg of PGF2alpha im 19 days after MGA (single) or 12.5 mg of PGF2alpha im 19 and 20 days after MGA (split). In Replication 3, heifers received the same treatments, with PGF2alpha initiated either 18 or 19 days after MGA. Estrus was detected for 72 h after PGF2alpha, with AI commencing 8-12 h after a detected estrus. Heifers not observed in estrus by 72 h were timed-AI concomitant with GnRH (100 microg im). Heifers from Replication 2 (n = 146) had blood samples collected at the initial PGF2alpha and at timed-AI to determine corpus luteum (CL) regression by evaluating plasma progesterone concentrations. The interval from MGA withdrawal to PGF2alpha did not have a significant effect on any variable in Replication 3 and there were no treatment by replication effects for any variables, therefore data were pooled. Modifying the PGF2alpha treatment from a single treatment to two treatments on consecutive days increased (P < 0.05) 72 h estrous response (43.2% versus 50.1%), timed-AI (23.9% versus 33.5%) and total-AI pregnancy rates (34.5% versus 42.5%), and CL regression (79.1% versus 92.5%), respectively. In Experiment 2, yearling Angus (n = 66) and 2-year-old BI (n = 68) heifers were synchronized as per Experiment 1 (with the initial PGF2alpha 19 days after MGA). Neither breed nor PGF2alpha treatment effected (P > 0.05) 72 h estrous response, total-AI pregnancy rate, or CL regression rate. In conclusion, treating yearling BI heifers with split treatments of PGF2alpha (given on two consecutive days) improved estrous response and pregnancy rates by increasing PGF2alpha-induced luteolysis.     

Uterine involution and postpartum ovarian activity in Nili-Ravi buffaloes

Uterine involution and postpartum ovarian activity were studied in 53 Nili-Ravi buffaloes. Mean intervals to uterine involution (26 days), regression of the corpus albicans of pregnancy (22 days), resumption of follicular activity (21 days) and first postpartum estrus (56 days) were not affected by the month of calving or age. Mean interval to formation of first corpus luteum (CL) after calving as indicated by progesterone in plasma (>/= 1.5 ng/ml) was 23.8 +/- 1.7 days, but only 52% of these CL were palpable. The number of CL formed before first postpartum estrus ranged from zero to five per buffalo; mean values based upon progesterone and palpation were 1.6 +/- 1.3 and 0.8 +/- 0.2, respectively. Based upon either progesterone or palpation, length of first postpartum luteal phase (7.9 or 6.6 days) was shorter than the luteal phase immediately preceeding the first estrus (12.1 or 8.9 days). Intervals from regular cyclic ovarian activity was not established until first estrus and intervals from the end of one luteal phase to the onset of the next were as long as three weeks. High concentrations of progesterone (>/= 1.5 ng/ml) on the day of behavioral estrus were seen in 23% of the buffaloes studied.